PSYCHO NEURO ENDOCRINO IMMUNOLOGY

PSYCHO NEURO ENDOCRINO IMMUNOLOGY

November 15th - 17th, 2001

Regensburg(Venue: Domspatzen-Gymnasium, Reichsstraße 22)

GE BIN

Meeting of the Volkswagen -

Foundation

and the GEBIN

(German Brain Immune Network)

Local Organising Committee

Rainer H. Straub Regensburg (Chairman*)

Manfred Schedlowski Essen (GEBIN)

*Address: Department of Internal Medicine I

University of Regensburg

D - 93042 Regensburg

Phone +49(0)941 944 7120

Fax +49(0)941 944 7121

Email [email protected]

Internet www.gebin.org

Scientific Committee

Hugo O. Besedovsky Marburg

Jan Born Lübeck

Adriana del Rey Marburg

Norbert Müller München

Thomas Pollmächer München

Peter Rieckmann Würzburg

Manfred Schedlowski Essen

Werner Scherbaum Düsseldorf

Günther Stalla München

Volker Stefanski Bayreuth

Rainer H. Straub Regensburg

Karlheinz Voigt Marburg

Congress Venue

St. Wolfgang - Saal des Musikgymnasiums der Regensburger Domspatzen

Programme Respective abstract numbers are given in parentheses. Thursday, November 15th, 2001 14:00 Opening of the conference 14:30 Neuroimmunology in the CNS

Chairs: R. Hohlfeld, U. Bogdahn (3, 6, 11, 14, 20, 21, 24, 29, 38, 40, 43, 55, 58, 62, 75, 82, 88, 93, 99)

16:30 Neuroimmune Endocrine Network in Psychiatric Disease

Chairs: T. Pollmächer, M.J. Schwarz (17, 18, 19, 36, 44, 45, 47, 48, 51, 64, 68, 70, 71, 72, 73, 84, 85, 92)

19:00 Evening Reception (with poster viewing session)

Friday, November 16th, 2001 09:00 Peripheral Neuroimmune Interactions

Chairs: D. Gemsa, C. Heijnen, R.E. Schmidt, J. Westermann (5, 10, 28, 30, 31, 32, 39, 46, 53, 56, 57, 63, 66, 76, 77, 78, 79, 80, 81, 86, 91, 97, 98)

12:00 Lunch (with poster viewing session) 13:00 Neuroendocrinology and Immune Function

Chairs: J. Born, S. Bornstein, A. Del Rey, E. Haen (4, 7, 15, 25, 26, 41, 50, 52, 54, 59, 61, 65, 69, 83, 87, 94, 95)

19:30 Party time in the Hotel Maximilian

Saturday, November 17th, 2001 09:00 Stress, Behaviour, and Immune Function

Chairs: R. Pabst, K. Schauenstein, M. Schedlowski, D. von Holst (1, 2, 8, 9, 12, 13, 16, 22, 23, 27, 33, 34, 35, 37, 42, 49, 60, 67, 74, 89,

90, 96) 12:30 Future aims of the GEBIN: Interactive discussion

Chairs: The Steering Committee of the GEBIN 13:00 Closing of the conference

A. Alipour, J. Faraji, Y. Jafair, A. Mirrezaie

Department of Psychology, Payame Noor University, Tehran, Iran

1. The effect of video games on children’s cortisol: A new perspective

Psychoneuroimmunology (PNI) is a new interdisciplinary field, researching the effects of psychological

factors on neural and immune systems, and their interactions. Most researchers believe that severe emo-

tions and chronic stressors often results in alteration of neural hormonal mediators, which are important

causes of stress–induced immune suppression. Furthermore, some working on PNI have attempted to

study the effects of severe emotions on cognitive (abstractive) activities of subjects. Due to controversial

data in this field, in order to study the effect of video games on hormonal alterations (cortisol), 16 students

with age average 12.6 yrs were selected randomly among a residential secondary school population.

They were assigned to four groups based on a Solomon design. Two experimental groups played com-

puter (video) games for one hour per day over 27 days. The blood samples were gathered based on

Solomon design and analysed in a laboratory for cortisol levels. The results were analysed statistically for

significant differences by Variance Analysis. The analysis indicated that the computer games increased

cortisol in the morning (p= 0.01) and in the evening (p= 0.04) significantly. The research finding have

implications for the effect of the computer games (as stressors) on children based on cortisol critical psy-

chophysiological influences.

P. Arck, M. Rose, R. Joachim, J. Niess, M. Hildebrandt, B.F. Klapp

Charité, Humboldt University Berlin, Germany

2. PsychoNeuroImmunology: Mission impossible ? Psycho-neuro-immunological (PNI) pathways have been suggested to be involved in the progression of

various diseases, i.e. inflammatory bowel disease, bronchial asthma or pregnancy complications. Many

insights have thereby been gained by basic laboratory work. In experiments employing laboratory animals

researchers could give their imaginativeness full scope in designing appropriate experimental stressors.

Delineating PNI pathways in humans is exceedingly difficult, and various approaches have been de-

signed to capture stress perception. Our procedure to evaluate stress perception in order to link these

observations to stress-triggered immune imbalances comprised the use of established stress question-

naires. The sensitivity and validity of these tests was proved by stress management interventions and

quality of life assessments. We were able to correlate immunological imbalances with high stress percep-

tion, i.e. the increased levels of abortogenic cytokine TNF and mast cells in decidua samples of patients

suffering from miscarriages, or the increase of the Th1 marker CCR5 in patients suffering from Crohn's

disease, an indisposition with high levels of inflammatory cytokines. We conclude that stress-triggered

effects on the progression of diseases might be identified using a questionnaire and linked to immu-

nological imbalances. Hereby, one must certainly be aware that correlation does not signify causality.

Nevertheless, future research is needed to fortify any correlation, enrolling a large number of individuals

preferably in a prospective study. Our recent data may therefore encourage physicians to use the per-

ceived stress questionnaires.

H.O.Besedovsky1, D. Balschun2, F. Pitossi3, H. Schneider1, W. Zuschratter2 and A. Del Rey1

1Institute of Physiology, Medical Faculty, Marburg, Germany; 2Leibniz Institute for Neurobiology, De-

partment of Neurophysiology, Magdeburg, Germany; and 3Institute for Biomedical Research, Fundacion

Campomar, Buenos Aires, Argentina

3. Endogenous IL-6 modulates the consolidation of hippocampal LTP IL-6, originally described as an immune-derived mediator, is also produced by brain cells and belongs to

the family of neuropoietins. Pharmacological evidence indicates that this cytokine can influence neuro-

endocrine functions, the synthesis, release, and turnover rate of several neurotransmitters, and behav-

ioural mechanisms integrated at brain levels. However, it is still unknown whether the production of IL-6 is

increased in the brain under non-pathological conditions and affects the activity of discrete populations of

brain neurons. We have found, in freely moving rats and in slices, that IL-6 gene expression is substan-

tially increased in the hippocampus during long-term potentiation (LTP), a process considered to underlie

certain forms of learning and memory. The increase in gene expression was long lasting, specific to po-

tentiation and could be prevented by inhibition of NMDA receptors. Blockade of endogenous IL-6 90 min

after tetanus, i.e. long after LTP started to consolidate, caused a remarkable prolongation of LTP. No

comparable effect was observed when the cytokine was blocked before and 5 minutes after tetanisation.

Thus, our data indicate that an increased local production of IL-6 during LTP may serve to control deci-

sive parameters involved in LTP consolidation and specificity. These modulatory actions of IL-6 and, as

we have already shown also of IL-1beta, may reveal a more general cytokine-neuronal mechanism that is

locally established in brain areas where neuronal activity is increased for a prolonged time. Supported by

the Volkswagen-Foundation and the SFB 297.

S. Birkmann, A. Schuld, M. Haack, W. Liebetrau, M. Dalal, T. Kraus, T. Pollmächer

Max-Planck-Institute of Psychiatry, Munich, Germany

4. Immunomodulatory properties of low dose dexamethasone - dose and time-of-day effects

The hypothalamo-pituitary-adrenal (HPA)-axis and the non-specific immune-response-system are closely

related. Glucocorticoids are important endocrinological modulators of cytokine release and increases of

granulocyte counts during pharmacological treatment with corticosteroids are well known. Furthermore, in

depressive patients it has been shown that a single, oral dose of dexamethasone decreased plasma lev-

els of inflammatory cytokines as interleukin (IL)-6, tumor-necrosis-factor (TNF)-alpha, and the soluble

TNF-receptor (sTNF-R) p75. However, it is still unclear, if those effects can be reproduced in healthy hu-

mans and whether there is circadian or dose dependent variability in these effects of glucocorticoids. In

order to explore these relationships, we assessed the immunomodulatory properties of a single, oral dose

of 1.5 or 3.0 mg dexamethasone at 9.00 or 21.00 hours in 43 healthy male volunteers. In this double-blind

placebo-controlled study blood was drawn on three consecutive days in the morning and evening. Differ-

ential blood cell counts were performed, and the immunocompetent cells were sub-specified by FACS-

analysis. Plasma levels of hormones, inflammatory cytokines and cytokine-receptors in plasma were dis-

played with commercially available ELISA and RIA. In all groups, dexamethasone had potent immuno-

modulatory properties, resulting in decreased levels of inflammatory cytokines and increased numbers of

granulocytes. Neither the applied dose of dexamethasone nor the circadian time of administration

seemed to have any influence on the results. The present study demonstrates that already small amounts

of glucocorticoids are potent immunomodulatory agents in healthy humans. In view of the strong circadian

modulation of immune parameters, it is somewhat surprising that time of day has no influence on dexa-

methasone-induced changes in cytokine secretion.

T. Brzoska, B. Altmann, T. Scholzen, D.-H. Kalden, A. Schwarz, T. Schwarz, S. Grabbe and T.A. Luger

Department of Dermatology and Ludwig-Boltzmann-Institute for Cell- and Immunobiology of the Skin,

University of Münster, Münster, Germany

5. Intravenous injection of the immunomodulator alpha-MSH induces hapten-specific tolerance

To investigate the underlying mechanisms, bone marrow derived dendritic cells (BMDC) were generated

with GMCSF/IL-4. On day 6, cells were treated with alpha-MSH (3h) and/or hapten (DNBS, 2h). After

washing, BMDC (500.000 cells) were injected i.v. into naive mice. 5d later, mice were challenged with

DNFB and ear swelling was measured. To evaluate tolerance induction, mice were resensitized after 14d

and rechallenged 5d later. Mice treated with DNBS-pulsed BMDC developed significant ear swelling upon

challenge with DNFB which was even enhanced after resensitization. In contrast, mice injected with al-

pha-MSH exposed BMDC showed significantly reduced ear swelling. Furthermore, ear swelling was still

suppressed following resensitization, indicating that tolerance had developed. Tolerance was hapten spe-

cific, since the same animals could be normally sensitised with the unrelated hapten oxazolone. In the

model of UV-induced tolerance it was recently shown that T-suppressor cells expressing CTLA-4 are

critically involved in mediating suppression. Therefore, BMDC were treated with DNBS and/or alpha-MSH

and co-incubated with lymph node derived T-lymphocytes. Alpha-MSH treated BMDC induced the gen-

eration of CTLA-4-positive subpopulations of CD4- and CD8-T-cells. Treatment (i.v.) of naive mice with

these CTLA-4-positive T-lymphocytes resulted in a significant inhibition of CHS and induction of toler-

ance. These data suggest that alpha-MSH pulsed DC may cause immunosuppression and tolerance in-

duction through the generation of CTLA-4-positive T-suppressor lymphocytes.

K. Bürger, H. Hampel

Department of Psychiatry, Dementia Research Section and Memory Clinic, Ludwig-Maximilian University

Munich, 80336 Munich, Germany

6. Expression of the interleukin-6 receptor complex in CSF and brain of pa-tients with Alzheimer's disease and controls

Interleukin-6 (IL-6) is a multifunctional cytokine serving as a relevant mediator of neuroregulatory and

inflammatory processes in the human central nervous system (CNS). We investigated the level of ex-

pression of IL-6 and its functionally related soluble (sIL-6R) and membrane bound receptor complex

(IL-6RC) in human CNS. The IL-6RC expression in brain tissue was investigated in different cortical re-

gions using semi-quantitative immunocytochemistry in AD patients and controls. Expression levels in

microglial cultures were analysed. After exposure to aggregated Abeta-peptide, conditioned medium from

the cultures was subjected to ELISA assays of IL-6. CSF-analysis: Sporadic AD and HC subjects were

analysed for IL-6, sIL-6R, soluble gycoprotein 130 (sgp130), and total tau levels. SIL-6RC concentrations

in CSF and plasma were correlated to polymorphisms of the IL-6 gene (IL-6vntr*C, IL-6prom*C). The IL-

6RC was differentially expressed by microglia, astroglia, neurons, and endothelial cells in both groups.

Microglia showed increased IL-6 in a dose-dependent manner after Abeta exposure, there was no differ-

ence between AD and HC microglia. In CSF, AD patients exhibited decreased concentrations of sIL-6R

and sgp130, but unchanged levels of IL-6. Combined examination of CSF sgp130 and tau protein maxi-

mised diagnostic separation between groups. Significant differences in plasma and CSF sIL-6R levels

were observed between AD patients and controls in non-carriers of the IL-6vntr*C allele (p = 0.001 and p

= 0.035, respectively). In subjects homozygous for the IL-6prom*C allele, AD patients had increased sIL-

6R levels in CSF and plasma compared to controls (p = 0.001). Our data further support the notion that

IL-6 plays a central role in inflammatory mechanisms involved in AD.

A. Buske-Kirschbaum, S. Fischbach, W. Rauh, J. Hanker, D.H. Hellhammer

Department of Psychology, University of Trier, Trier, Germany

7. Increased responsiveness of the hypothalamus-pituitary-adrenal (HPA) axis to stress in newborns with atopic disposition

In previous studies atopic patients showed attenuated cortisol responses to psychosocial stress which is

suggestive of a hyporeactive hypothalamus-pituitary-adrenal (HPA) axis in this patient group. Regarding

the anti-inflammatory role of glucocorticoids, reduced responsiveness of the HPA axis under stress may

be one potential explanation of stress-induced exacerbation of atopic symptoms. The present study

evaluated whether hyporeactivity of the HPA axis is a feature related to the disposition of atopy rather

than a consequence of an ongoing chronic allergic inflammatory process. Newborns with an atopic dispo-

sition (parental atopy; n=31) and without atopic disposition (no parental atopy; n=20) were recruited. To

further assess atopic disposition, total IgE levels were determined in the cord blood of the neonates.

Three days after birth, a blood sample was obtained by a heel prick which is part of a standard pediatric

examination. Blood sampling by heel prick is well known to be a significant stressor resulting in activation

of the HPA axis in newborns. Analysis of salivary cortisol indicated a significant increase of cortisol levels

in the newborns after the stressor with a trend towards an elevated cortisol response in babies with a

family history of atopy or with elevated levels of cord IgE (>/= 0.5 kU/L). Neonates with a positive parental

atopic heritage and elevated cord IgE were found to show significantly elevated cortisol responses to the

heel prick stress when compared to newborns without a parental history and nomral cord IgE values.

Moreover, cord IgE levels were significantly correlated with basal cortisol levels and the cortisol response

to the stressor. These findings suggest that atopic disposition in neonates is associated with altered re-

sponsiveness of the HPA axis to stress which may increase the vulnerability to develop manifestation of

atopy in later life.

L. Dawils, V. Stefanski

Department of Animal Physiology, University of Bayreuth, Bayreuth, Germany

8. Social confrontation and tumor metastasis in male laboratory rats: health consequences of social stress

Many studies on social stress suggest negative consequences for immune functioning and health. In-

creased levels of adrenal hormones are usually thought to mediate this effect. However, convincing evi-

dence for a causal relationship between hormonal action, immunosuppression and disease outcome un-

der naturalistic stressful conditions is still rare. To address this question, we investigated the conse-

quences of continuous social confrontations (three hours as well as two days) on the adrenomedullary

activity and the antimetastatic activity of natural killer cells, using the MADB tumor model. Adrenal intact

subdominant males showed an about ten-fold increased tumor load in the lungs 24 hours after tumor

injection compared to undisturbed controls. Adrenal tyrosine hydroxylase (TH) activity increased in loser

males after two days of confrontation, elevated levels of plasma catecholamines were correlated with the

TH activity. The number of tumor cells retained in the lungs was positively correlated with submissive

behaviour as well as adrenomedullary activity. These findings demonstrate that social defeat not only has

negative consequences to metastatic development per se, but also affects the disease outcome depend-

ing on the quality of subordination. In addition, adrenal demedullation reduced the number of tumor cells

in the lungs of loser rats by 50%. This emphasises the involvement of the sympathetic-adrenomedullary

axis in mediating the suppression of in vivo natural killer cell activity. Supported by the Volkswagen Foun-

dation (I/75144)

R. Deinzer, B. Waschul, N. Granrath, A. Herforth, R Stiller-Winkler, H. Idel

Institute for Medical Psychology, Department of Periodontology, Institute for Hygiene, Heinrich-Heine-

University Düsseldorf, Düsseldorf, Germany

9. Stress and periodontitis - current perspectives

Periodontitis is an inflammatory disease with a prevalence of about 15-50% among western communities.

The major characteristics of this disease are degradation of alveolar bone and periodontal tissues in-

duced mainly by the inflammatory immune response. Interleukin 1beta is thought to be a key cytokine in

the etiopathogenesis of the disease. Several studies during the last related psychological stress to perio-

dontal breakdown. Indeed, major life events as well as stressful life circumstances seem to accelerate the

disease's progress. The mechanisms behind these stress-periodontitis-relationships are, however, not

well understood. Both, antigen load as well as immune reponses to periodontal antigens as well as other

risk factors like smoking or nutrition might mediate between psychological stress and periodontitis. In our

group we analyse stress effects on putative behavioural and immunological mediators in humans. Thus,

we analyse stress effects on oral hygiene behaviour, the resulting antigen load and on local immune pa-

rameters like interleukin 1beta, interleukin 1 receptor antagonist, prostaglandin E2, and immunoglobulin A

in the gingival crevice. Most of our studies indicate stress induced alterations implicating an increased

periodontal risk. However, some less clear-cut results suggest that nature and duration of the stressor as

well as time of assessment play an important role. In this presentation we will summarise our present

research on stress-periodontitis relationships and discuss current research perspectives (supported in

part by DFG De527/3-1).

Del Rey1, A. Kabiersch1, E. Roggero1, M. Schäfer2, E. Weihe2, H.O. Besedovsky1

1Department of Immunophysiology, 2Institute of Physiology, and Institute of Anatomy, Medical Faculty,

Marburg, Germany

10. Interactions between the immune and the sympathetic nervous system dur-ing a genetically-determined autoimmune lymphoproliferative disease

Mice homozygous for the lpr gene develop an autoimmune proliferative disease that resembles certain

characteristics of systemic lupus erythematosus in humans. Lpr/lpr mice do not express Fas, and thus

apoptosis cannot be triggered through this receptor. Because we have recently found that noradrenaline

(NA), the main sympathetic neurotransmitter, can induce apoptosis in lpr/lpr lymphoid cells, we have be-

gan to study the sympathetic innervation in lymphoid organs of these mice. We have found that the con-

centration of NA in the spleen of C57Bl/6J lpr/lpr mice markedly differs from that of normal animals. This

difference is already observed early in ontogeny. A profound decrease in splenic NA concentration, with

reduction in the density of tyrosine hydroxylase-immunoreactive fibres and disappearance of fine proc-

esses in parenchymal nonvascular localizations, is observed at relatively advanced stages of the disease.

We also found that there is an inverse correlation between IgM blood levels and NA concentration in the

spleen when the disease is overtly manifested. Preliminary results indicate that sympathetic denervation

at birth differentially affects IgM blood levels in normal and lpr/lpr mice. Interestingly, about 40% of the

sympathectomized lpr/lpr male mice died within 6-8 weeks. As a first pharmacological approach, a small

group of young lpr/lpr female mice was chronically treated with a beta-blocker. This treatment resulted in

significantly increased IgM blood levels when compared to those of the control group. These findings

support the hypothesis that alterations in the sympathetic nervous system of lpr/lpr mice may contribute to

the pathogenesis of the autoimmune disease. Supported by the Volkswagen-Foundation and the SFB

297.

K. Dinkel, M. Rickert, G. Möller1, J. Adamski2, H.-M. Meinck3, W. Richter

1Department of Orthopedic Surgery, University of Heidelberg, Germany, 2Department of Mammalian

Genetics, 2GSF-National Research Center of Environment and Health, Neuherberg, Germany,

3Department of Neurology University of Heidelberg, Germany

11. Identification of 17ß-Hydroxysteroid Dehydrogenase Type 4 as a novel 80 kDa Antineuronal Antigen in Stiff-Man-Syndrome

Stiff-man syndrome (SMS) and its variants, Stiff leg syndrome and progressive encephalomyelitis with

rigidity and myoclonus are rare disorders of the central nervous system. Autoantibodies to the enzyme

glutamic acid decarboxylase (GAD) are present in 60-80% of these patients. Several studies indicate that

autoimmunity against GAD may be only part of a multiantigenic immune response which may also affect

the endocrine system. In a patient newly diagnosed with SMS we screened the peripheral B cell reper-

toire for antibodies reactive with rat cerebellum. We isolated stable human monoclonal B cell lines with

staining patterns distinct from GAD antibodies, and further characterised these antibodies. Amongst five

antibodies with brain reactivity, one human monoclonal antibody (mAb CB11) reacted with both Purkinje

cells and ependymal cells and precipitated an 80 kDa protein from radiolabelled rat neuronal primary

cultures. An 80 kDa reactivity was also seen with 12% (3/25) of SMS sera and 13% (2/15) of SMS cere-

brospinal fluid (CSF) samples. The 80 kDa antigen was affinity-purified with CB11 from ependymal cell

enriched neuronal cultures and its sequence identified by mass spectrometry as 17b-hydroxysteroid de-

hydrogenase type 4 (17ß-HSD4). This peroxisomal enzyme is important for steroid inactivation and b-

oxidation of fatty acids. MAb CB11 immunoprecipitated steroid dehydrogenase enzyme activity from rat

liver extracts and reacted also with recombinant rat 17ß-HSD4. In conclusion we here identified 17ß-

HSD4 as a possible novel target of autoimmunity in SMS supporting the hypothesis that the many differ-

ent facultative symptoms seen in individual SMS patients may occur on a heterogeneous autoantibody

background.

M. Ebrecht, A. Buske-Kirschbaum, S. Kern, C. Kirschbaum, D.H. Hellhammer

Department of Psychological Medicine, Guy's King's and St Thomas' School of Medicine, Kings College,

London, UK

12. Enhanced catecholaminergic reactivity and lymphocyte redistribution after acute stress in patients with psoriasis

Exacerbation of various skin disorders has been documented after phases of severe stress in humans.

This study investigated the mechanisms of how a short-term psychosocial stressor can affect lymphocyte

subpopulations in psoriasis. Subjects with the Th1-type inflammatory skin disorder psoriasis (n=23, mean

age 32.6) and healthy controls (n=25, mean age 28.5) were confronted with a psychosocial laboratory

stressor (Trier Social Stress Test, TSST) consisting of public speaking and mental arithmetic. Blood and

saliva samples were repeatedly obtained over a 2h period for determination of cortisol, ACTH, cate-

cholamines and leukocyte subsets in peripheral blood. Insignificant group differences in basal and stress

induced cortisol and ACTH responses and in dexamethasone-induced cortisol suppression suggest that

the HPA axis is unaffected in psoriasis. However, differential stress responses were observed in redistri-

bution of monocytes and lymphocyte subsets. While peripheral monocytes and CD4+ cell counts were

significantly elevated in the psoriasis group immediately after the stressor, no such changes could be

observed in the control group (Monocytes: F=2.75, p=.046; CD4+ cells: F=3.09, p=.030). Monocytes and

CD4+ cells play an important role in inducing hyperproliferation of keratinocytes in psoriasis. Furthermore,

stress induced secretion of the catecholamines adrenaline (F=4.32, p=.006) and noradrenaline (F=

3.11,p=.029) was elevated in the psoriasis patients. Positive correlations between adrenaline elevation

and increase in monocyte counts (r=.37, p=.016) as well as noradrenaline and CD4+ cells (r=.38, p=.014)

were found. The contribution of personality factors and stress susceptibility to the alterations in the immu-

nological and endocrine stress profile in psoriasis will be discussed.

F. Eggert

Department of Psychology, University of Kiel, Kiel, Germany

13. Behavioural specificity of MHC-associated olfactory preferences

The major histocompatibility complex (MHC) is a gene cluster that is involved in antigen recognition and

tolerance induction in the anticipatory immune system of vertebrates. The most prominent feature of the

MHC, its outstanding genetic diversity seems to be selectively maintained in contemporary natural popu-

lations by parasite- and pathogen-driven selection, namely negative frequency dependent selection and

overdominance. However, the high degree of heterozygosity found in natural populations of most species

seems to be promoted not only by overdominance but also by non-disease-based selection such as mat-

ing preferences. Results of a number of studies confirm that MHC-associated olfactory signals exist in

mice and rats. In mice these signals contribute to imprinted mating preferences which serves to promote

heterozygosity. The aim of our recent studies was to determine whether these olfactory preferences are

confined to mating situations. We examined the olfactory preferences of inbred mice choosing between

the scents of genetically different inbred strains with known MHC type in a standard non-mating situation

(preference test). The results show that MHC-associated olfactory preferences in mice exert their influ-

ence on behaviour within the mating situation but not in standard preference tests. (This and related work

was supported by the Volkswagen-Foundation)

U. Eisel, L. Marchetti, V. Fontaine, M. Klein, K. Pfizenmaier

Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart, Germany

14. Neuroprotective Function of TNF is mediated via Akt/PKB signalling pathway We have investigated the neuroprotective function of TNF in different transgenic and knockout mouse

models and in in vitro studies. In a model of retinal ischaemia TNFR1 knockout mice were completely

protected and showed Akt/PKB phosphorylation 6 hours after ischaemia. TNFR2 knockout mice showed

increased lesions, suggesting that TNFR2 signalling has a protective function in ischaemia. Using an in

vitro model with primary cortical neurons we used glutamate induced excitotoxicity to study the function of

TNF in neuronal injury. Cortical neurons from the transgenic mouse line tg41.3IZI, a mouse line express-

ing TNF under control of the murine NMDA receptor subunit NR2B promoter in the cortical and hippo-

campal neurons, were completely protected from glutamate induced excitotoxicity, whereas cortical neu-

rons from wild type C57Bl/6 were highly sensitive to glutamate. Cortical neurons from wild type mice pre-

treated with soluble human or murine TNF were also protected from excitotoxicity and showed NFkappaB

activation within 20 to 40 minutes after treatment with glutamate. MK-801, a NMDA receptor antagonist,

abolished also excitotoxicity. Treatment with either TNF or glutamate alone showed very little effects on

Akt/PKB phosphorylation. However, treatment with both TNF and glutamate together induced strong

Akt/PKB phosphorylation. These results suggest a cross talk of TNF receptor signalling and NMDA recep-

tor signalling in glutamate induced neuronal injury and Akt/PKB phosphorylation.

S. Elsenbruch, D. Özcan, A. Lysson, G. Holtmann, M. Schedlowski

Department of Medical Psychology and Department of Gastroenterology and Hepatology, University

Clinic of Essen, Essen, Germany

15. Neuroendocrine-immune responses to food stimulation in patients with irri-table bowel syndrome (IBS) and controls

It is well-established that IBS patients show altered neuroendocrine responses to various visceral stimuli,

and recent data support the role of infectious processes in pathophysiology of IBS. The goal of this study

was to characterise changes in neuroendocrine and immune parameters before and after a food chal-

lenge. Blood samples were drawn from 10 women with IBS and 10 age-matched healthy women before

(fasting), and 20, 45, 75, and 105 minutes following a standardised meal containing 500Kcal. Prolactin,

cortisol, ACTH, and growth hormone, cardiovascular responses, peripheral lymphocyte subpopulations,

and cytokine production (IL-6 and TNF) by blood monocytes were analysed before and after the meal.

Compared to controls, patients showed significantly enhanced heart rate (p<.05) and blood pressure re-

sponses (p<.01) to the meal. In addition, patients demonstrated a significantly greater postprandial in-

crease in norepinephrine, and a significantly diminished ACTH response (both p<.05). Analyses of im-

mune parameters revealed a significantly enhanced increase in the number of peripheral granulocytes,

and a significantly diminished response of monocytes and CD3+ T cells in patients compared to controls

(all p<.05). Postprandial changes in LPS-stimulated TNF and IL-6 concentrations were also significantly

diminished in patients. These results support that IBS patients respond to a food challenge with abnormal

autonomic and HPA-axis activation, which is accompanied by alterations in cellular immune responses

suggesting disturbances of the neuroendocrine-immune axis in IBS.

H. Engler, S. Kurth, S. Hoves, V. Stefanski


16. Social stress and immunomodulation: specific role of glucocorticoids and catecholamines

Acute and chronic social stressors have been shown to influence the distribution and function of immune

cells in blood as well as in central and peripheral lymphoid organs of laboratory rats. These immunologi-

cal alterations were associated with an activation of the hypothalamus-pituitary-adrenocortical and the

sympathetic- adrenomedullary system, resulting in elevated blood levels of free glucocorticoids (GC) and

catecholamines. Since both, cells of the lymphoid and the myeloid cell lineage express receptors for

these hormones, this study was conducted to elaborate the endocrine mechanisms that were responsible

for immunomodulations in socially stressful situations. We used adrenalectomised (ADX) as well as GC

type-II receptor antagonist (RU486) treated male laboratory rats in social confrontation experiments to

investigate the specific impact of adrenal-derived hormones on immune cells in blood and thymus. Adre-

nalectomy as well as RU486 treatment abolished the stress- associated immune changes in the thymus

(increased T-cell apoptosis, reduced cellular proliferation), but affected only partially the alterations in the

peripheral blood: the decrease in lymphocyte numbers (Thelper, CTL, and B cells) was completely elimi-

nated by ADX and RU486 whereas the typical increase in NK cells (+170%) and neutrophils (+260%) was

unaffected by these manipulations. This supports the view that stress-associated alterations in mature

and immature lymphocytes were mediated by GCs acting on type-II adrenal steroid receptors whereas

the increase in NK cells and neutrophil granulocytes has to be mediated by adrenal-independent

mechanisms, e.g. via stimulation of adrenergic receptors by norepinephrine. Supported by the Volks-

wagen Foundation (I/75144)

B. Fellerhoff, B. Bondy, N. Müller, M. Vogel, B. Laumbacher, R. Wank

Institute for Immunology, Ludwig-Maximilians University Munich, Munich, Germany

17. HLA factors in families and unrelated patients with psychiatric disorders: significant deviations in recombination frequencies and allelic frequencies of TAP but not LMP alleles

Analysis of HLA factors in 17 families with 18 patients with schizophrenic, 21 patients with bipolar or uni-

polar disorders and 11 patients with other disorders showed no significant Lod score between HLA and

any disorders or combinations of disorders. However, the frequency of recombination between HLA-A

and HLA-B was higher than expected (Fisher's exact p = 0.012). Further studies are required to verify

extent and nature of these events. To find a possible common pathogenic denominator of the disease, we

expanded our investigations in the HLA complex and focused on polymorphisms of TAP and LMP genes,

involved in processing of MHC-I-presented antigens. ARMS-PCR was used to investigate the polymor-

phisms of the LMP2 and LMP7 genes in 165 patients and 165 controls. The frequency of the LMP2-

Arg/LMP7-Gln/LMP7-Lys genotype was increased in patients, but did not reach the significant level after

correction. We found significant alterations in allele and genotype frequencies looking at TAP1 polymor-

phisms by sequence analysing the DNA of 135 schizophrenic patients and 162 controls. The

TAP1*A1/TAP1*B1 genotype was significantly increased (p = 0.00274) in the patients group whereas the

TAP1*A1/TAP1*B4 genotype was reduced. We calculated the relative risk and found that individuals with

the TAP1*A1/TAP1*B1 genotype have a 10 fold risk to develop schizophrenia, whereas persons with the

TAP1*A1/TAP1*B4 genotype have a 9.8 fold reduced risk. Because TAP*B1 and TAP1*B4 differ in the

predicted peptide binding region of the TAP1 molecule, we suggest that selection of transported peptides

occurs and finally the presentation of the peptides could be altered in schizophrenics.

S. Fest, A.C. Zenclussen, H. Lange, C. Weber, B. Klapp, P.C. Arck

Charité, Humboldt University, Berlin, Germany

18. Downregulation of Th1-markers by progressive muscle relaxation (PMR)

Stress is known to influence the immune, nervous and endocrine systems, but very little insight has been

gained in this complex network. It remains still unclear whether stress might rather have a "Yin" or a

"Yang" effect on the physiological homeostasis. Furthermore, it has been reported that stress manage-

ment can be improved by PMR. Here, we investigated the presence of CCR5+ cells and the intracellular

production of INF-gamma and TNF in immunocompetent cells under PMR. PMR has been applied over

10 weeks (n=5). At the beginning and the end blood samples of PMR-volunteers and a non-PMR-group

(n=5) were taken. The presence of CCR5+, INF-gamma or TNF producing immunocompetent cells were

evaluated by flow cytometry. The stress was assessed by using a Perceived Stress Questionnaire (PSQ)

and a Perceived Stress Scale (PSS). CCR5+ cells and the amount of INF-gamma and TNF was signifi-

cantly reduced after PMR treatment. CD57+ NK cells seemed to be the major source of INF-gamma,

whereas TNF was produced by CD3+ cells. No changes in of immune parameters were present in the

control group. The PSS was decreased with PMR when it remained unchanged in control individuals at

the two assessment points. Decreased Th1 cytokines after PMR reflects how influential stress interven-

tion may be on the physiological homeostasis. Thus, it might be useful to improve stress management

capacities for the progression of several diseases. Remarkably, CCR5 might be a sensitive marker of

stress measurement.

B.L. Fiebich, B. Müksch, M. Böhringer, M. Hüll

Department of Psychiatry and Psychotherapy University of Freiburg, Freiburg, Germany

19. Neuroinflammatory circuits in Alzheimer's disease: Interleukin-1 induces cyclooxygenase 2 in human neuroblastoma cells

Cyclooxygenase 2 (COX-2) has been shown to be overexpressed in neurons in Alzheimer's disease

(AD). In the normal brain, neuronal COX-2 expression is regulated by transsynaptic stimulation via

NMDA-receptors. This pathway of neuronal COX-2 induction can not explain neuronal COX-2 overex-

pression in AD due to the loss of synaptic stimulation in AD. Therefore we investigated other potential

stimulators of neuronal COX-2 expression. The expression of interleukin-1 (IL-1) in microglia is one of the

early and consistent signs of microglial activation in AD. Here we report that IL-1 induces COX-2 in the

human neuroblastoma cell line SH-N-SK which is accompanied by neuronal prostaglandin E-2 (PGE-2)

production. The induction of COX-2 in human neuroblastoma cells involves the activation of p38 mitogen-

activated kinase (p38 MAPK) and nuclear factor-kappaB (NFkappaB). Previous neuropathological data

showed an activation of p38MAPK and NFkappaB in neurons in AD. Therefore our results supports a

model of neuroinflammation in AD in which microglial IL-1 synthesis is followed by neuronal p38 MAPK

activation and NFkappaB activation which both are involved in subsequent COX-2 expression and PGE-2

production. Neuronal expression of COX-2 may lead via not yet well described intracellular pathways to

neuronal cell death. Neuronal release of PGE-2 may stimulate further astroglial activation including inter-

leukin-6 synthesis. Neuroinflammatory circuits may be interrupted by drugs that reduce IL-1 induced neu-

ronal COX-2 expression.

B.L. Fiebich, S. Schleicher, O. Spleiss, M. Czygan, M. Hüll

Department of Psychiatry and Psychotherapy University of Freiburg, Freiburg, Germany

20. Neuroinflammatory circuits in Alzheimer’s disease: Prostaglandin E-2 in-duces astroglial interleukin-6 synthesis independent of cAMP formation

In Alzheimer's disease (AD) interleukin-6 (IL-6) is present in early amyloid plaques and elevated levels of

IL-6 are measured in brain extracts. We have previously shown that prostaglandin E-2 (PGE-2) is able to

induce IL-6 in the human astroglioma cell line U373 MG and in primary rat astrocytes. In many different

mesenchymal cell lines the induction of IL-6 by PGE-2 is regulated via the activation of the prostaglandin

receptor EP2 with subsequent elevation of intracellular cAMP. In this study we show that PGE-2-induced

IL-6 secretion in human astroglioma cells is independent of EP2 activation or rise of intracellular cAMP

despite the fact that PGE-2 induces a rise in intracellular cAMP via EP2 receptors in these cells. In con-

trast to the expected involvement of EP2 receptors and cAMP we could demonstrate that IL-6 secretion

depends on the activation of protein kinase C (PKC) and p38 mitogen activated protein kinase

(p38MAPK). Preliminary pharmacological experiments with agonists and antagonists suggest that the

astroglial IL-6 secretion depends on a non EP1/EP2/EP3-like PGE-2 receptor. The reduced tolerability of

COX-inhibitors in old age has stimulated the research for prostaglandin receptor antagonists for anti-

inflammatory treatment. Our study suggests that in human glial cells a yet not characterised PGE-2 re-

ceptor with a link to PKC / p38MAPK activation is crucial for glial activation in neuroinflammation in AD.

P. Flachenecker, T. Kümpfel1, M. Gottschalk1, P. Rieckmann, F. Weber1, C. Trenkwalder1, K.V. Toyka

Department of Neurology, Julius-Maximilians-Universität Würzburg, Germany, and 1Max-Planck-Institute

for Psychiatry, Munich, Germany

21. Autonomic dysfunction in patients with multiple sclerosis and fatigue

Fatigue is one of the most common disabling symptoms in patients with multiple sclerosis (MS), but the

pathophysiological mechanisms remain to be determined. In this pilot study, we evaluated autonomic

function in MS patients suffering from fatigue. Standard autonomic function tests (AT) for parasympathetic

(heart rate variability to Valsalva manoeuvre, deep breathing and active change of posture) and sympa-

thetic function (blood pressure responses to active change of posture and sustained handgrip), power

spectrum analysis of heart rate variability (PSA), and resting levels of serum catecholamines (norepineph-

rine, NE, and epinephrine, EPI) were assessed in eight female MS patients, age 37 ± 7 years with fatigue

(Fatigue Severity Scale 5.2 ± 0.7). Results were compared to those obtained in eight age- and sex-

matched healthy controls. The parameters of AT and PSA were similar in both groups, as were NE levels.

However, EPI levels were significantly reduced in MS patients with fatigue compared to healthy volun-

teers (17 ± 4 ng/l vs 24 ± 6 ng/l, p < 0.025, t-test). The results of this pilot study suggest that sympathetic

activity may be reduced in MS patients with fatigue. However, larger studies in patients with and without

fatigue are warranted to clarify whether autonomic dysfunction contributes to MS-related fatigue.

M.U. Göbel, Yu Fen Xie, A.E. Trebst, J.F. Steiner, M.S. Exton, S. Freede, A. Canbay, U. Heemann, M.C.

Michel, M. Schedlowski

Departments of Medical Psychology, Physiology and Medicine, University of Essen, Germany

22. Behaviourally conditioned immunosuppression

One of the most impressive demonstrations of brain to immune system interaction is classically condi-

tioned immunomodulation. Our group has established a rat model of conditioned immunosuppressive

effects to examine the mechanisms and biological relevance of this protocol. By using cyclosporin A

(CsA) we showed that classical conditioning reduces splenocyte proliferation and both interleukin (IL)-2

and interferon (IFN)-γ production. These effects were driven by neural innervation of the spleen, as surgi-

cal denervation completely blocked the conditioned effects. Moreover, we demonstrated that our protocol

inhibits contact sensitivity and prolongs the survival time of transplanted heart allografts in rats, thereby

demonstrating the biological relevance of conditioned immunosuppression. Further, a subtherapeutic

dose of CsA extended the graft survival time in conditioned animals, showing a synergistic effect of the

subtherapeutic drug with the conditioning regimen. These data thus suggest that conditioned immuno-

suppression may have utility in a clinical setting. Therefore, in a first step to examine the clinical applica-

bility of this, we examined the conditionability of the immunosuppressive effects of CsA in healthy hu-

mans. This study was conducted as a double blind, placebo controlled experiment, and results from the

ongoing analyses will be presented.

J. Götze, D. Goldmann, J. Niess, A. Dignass, P. Arck

Charité, Humboldt University Berlin, Germany

23. Influence of stress on immune-competent cells in patients suffering from ulcerative colitis

Stress is known to influence the pathophysiology of inflammatory bowel disease (IBD). Recent data indi-

cate that perceived stress could exacerbate ulcerative colitis in human and mice (Levenstein, 2000;

Qui,1999). Experimental studies in mice employing TNBS-induced colitis exposed to stress provided

some insights into the underlying effects of stress in intestinal inflammation. This suggested an involve-

ment of immune-competent cells, such as CD4+/CD8+ T-cells. In order to probe the validity of the con-

clusions drawn from murine experiments in human IBD, the following study was performed. We measured

the stress score and colitis activity index of patients suffering from ulcerative colitis using an established

questionnaire. The presence and distribution of peripheral immune-competent cells, such as, CD56+ or

CD57+ NK-cells, CD3+, CD4+ and/or CD8+ T-cells, CCR3+ and CCR5+ cells were determined by flow

cytometry. Serum levels of IL-6 and IL-10 were analysed by ELISA. The local presence of T cells, NK

cells and mast cells at the mucosal site was confirmed by IHC. We observed a significantly negative cor-

relation between systemic CD3+/CD57+, CD4+/CD57+, CD8+/CD56+, CD3+/CD56+, serum IL-6 level

and the stress score, and a significantly positive correlation between CD8+cells and stress locally. The

percentage of chemokine-receptor expressing cells and the serum IL-10 levels was not sensitive to stress

perception. We conclude that using a questionnaire to score perceived stress in human might be an ap-

proach to generate non-biased stress levels. As presented in this study, we could show that T-NK cells

are negatively correlated with high levels of perceived stress in our patient cohort.

M. Gottschalk, T. Kümpfel, A. Yassouridis, F. Holsboer, F. Weber

Max-Planck Institute of Psychiatry, Section of Neurology, Munich, Germany

24. Fatigue and regulation of the hypothalamo-pituitary-adrenal axis (HPA axis) in multiple sclerosis (MS)

Several neuroendocrine studies have suggested alterations of the HPA axis in patients suffering from

chronic fatigue syndrome. In recent studies we demonstrated a hyperreactivity of the HPA axis in MS

patients, which correlated to the course and stage of the disease. 44 untreated patients (31 women and

13 men) with clinically-definite MS, age 39.2 ±1.5 years (mean ± SEM) and disease duration of 7.5 ± 1.0

years were studied. 31 patients suffered from relapsing-remitting (RR) and 13 from a progressive course

(CP) of MS. Mean EDSS according to the Kurtzke´s scale was 2.7 ± 0.3. Subjects were interviewed and

asked to fill in fatigue questionnaires: Visual Analogue Scale (VAS), Fatigue Severity Scale (FSS) and

Modified Fatigue Impact Scale (MFIS). For assessing HPA axis activity the combined Dex-CRH test was

performed. We found a significant correlation between the severity of fatigue (cluster-analysis of the vari-

ables VAS, FSS and MIFS) and the HPA axis-activity (AUCACTH: area under the curve, MLACTH: mean

location of the curve): patients with fatigue showed higher ACTH-levels (AUCACTH: 49.6 ± 6.8 arbitrary

units (AU), MLACTH: 11.0 ± 1.5 pg/mL, 22 patients) than patients without (AUCACTH: 30.8 ± 3.2 AU,

MLACTH: 7.1 ± 0.7 pg/mL, 19 patients), p<0.05 (univariate F-tests). Cortisol levels showed a trend.

Therefore our results demonstrated a stronger hyperreactivity of the HPA axis in MS patients with fatigue

than in MS patients without fatigue as demonstrated by elevated AUCACTH and MLACTH. Proinflamma-

tory cytokines may be the cause of HPA axis alterations and a stronger feeling of fatigue.

V.V. Grinevich1,2, G.F. Jirikowski1 1Department of Anatomy II, Friedrich Schiller University Jena, Jena, Germany; 2Department of Histology

and Embryology, Russian State Medical University, Moscow, Russia

25. Magnocellular hypothalamic-neurohypophyseal system under acute inflam-mation

Since stimulatory effects of several cytokines on activity of the hypothalamic-neurohypophyseal system

(HNS) have been well documented, data regarding to reaction of this system to acute inflammation were

quite controversial. Therefore we aimed to study effects of systemic inflammation, induced by bacterial

endotoxin lipopolysaccharide (LPS) on HNS with special reference to its vasopressin (VP) compartment.

Moreover we studied responsiveness of this system to osmotic stimulation in rats receiving 250 micro-

gram per 100 g BW of LPS, 3 or 6 h prior to ip injection of hypertonic saline 900 mM NaCl (ipHS). LPS

injection had no effects on plasma vasopressin (VP) levels in control rats but it potentiated the responses

to ipHS independent from plasma sodium levels and hemodynamic changes. High magnification light

microscopy and electron microscopy examination of the PP shows that LPS injection induces a significant

increase in the proportion of large terminals, some of which appeared as gigantic swelling. Osmotic

stimulation reduced the total number of VP labeller terminals and intensity of staining, that were more

profound in LPS pretreated rats. There were no changes in oxytocin terminals in the PP. In conclusion the

findings reveal an anatomical substrate for increased releasable pools of VP and potentiation of osmotic-

stimulated VP secretion observed during LPS-induced acute inflammation in the rat.

M. Haack, T. Kraus, A. Schuld, M. Dalal, D. Köthe, T. Pollmächer

Max Planck Institute of Psychiatry, Munich, Germany

26. Diurnal variations of interleukin 6 plasma levels during sleep and wakeful-ness: effects of particularities of blood drawing

Recent findings suggest that inflammatory cytokines are involved in sleep regulation. In part, this idea is

based on studies showing that systemic levels of interleukin-6 (IL-6) are affected by sleep and sleep dep-

rivation. However, i.v. catheters used for repetitive blood sampling were reported to increase local IL-6

production, which might confound sleep-dependent or circadian changes in the systemic production of

this cytokine. To further examine the effects of blood drawing procedures on IL-6 plasma levels, 12

healthy young male subjects participated in a 24-hour cross-over study protocol involving sleep and sleep

deprivation. Blood was collected half-hourly through an i.v. line and one additional sample was taken by a

simple needle stick from the contralateral arm in parallel to the last sample from the catheter. Difficulties

in blood sampling, the plasma levels of IL-6, cortisol and subjective sleepiness were quantified. In sam-

ples from the i.v. line there was a linear increase in IL-6 levels in both conditions, whereas the amount of

IL-6 detected in the needle stick sample at the end did not differ from baseline. IL-6 levels were signifi-

cantly higher in samples rated as difficult, and those difficulties were more frequent during sleep com-

pared to nocturnal wakefulness. IL-6 levels did not correlate to variations in cortisol levels of subjective

sleepiness. We conclude that variations in IL-6 plasma levels measured in samples from an i.v. catheter

are caused by changes in local cytokine production rather than by physiological changes in circulating IL-

6 levels.

P. Haake, M.U. Göbel, K. Heberling, S. Elsenbruch, M. Schedlowski, T. Krueger

Institute for Medical Psychology, University Clinics of Essen, Essen, Germany

27. Orgasm-induced circulation of lymphocyte subsets

In a series of studies we have analysed the psychoneuroendocrine response to sexual arousal in hu-

mans, showing that orgasm stimulates cardiovascular activity, catecholamine secretion and induced a

pronounced increase in plasma prolactin concentrations. In this ongoing study we investigate the effects

of masturbation-induced orgasm on immunological parameters in healthy young males. In a cross-over

design, eight participants completed an experimental session, where a documentary film was observed

for 20 min, followed by 20 min of a pornographic film, and a further 20 min documentary. Subjects also

participated in a control session, where the participant watched a documentary film for 60 min. Following

10 min of pornographic film in the experimental session, subjects were asked to masturbate until orgasm.

Sexual arousal was estimated by different questionnaires. Cardiovascular parameters were monitored

continuously. Blood was drawn before, and both 5 min and 30 min after orgasm for determination of leu-

kocyte subsets and lymphocyte subpopulations. Paralleling elevated sexual arousal, orgasm induced

transient increases in heart rate and blood pressure and led to significant increases in the number of

CD3+ lymphocytes and CD3-CD16+CD56+ lymphocytes in comparison to the control session. In contrast,

CD3+CD4+ lymphocytes were not affected. These preliminary data of this ongoing study indicate that

orgasm induces acute changes in lymphocyte circulation.

E. Haen, M. Buchberger, W. Dorsch

Department of Clinical Pharmacology/Psychopharmacology and Clinic/Polyclinic for Psychiatry, University

of Regensburg

28. Bronchodilatation by antidepressants in allergen sensitised guinea pigs

Tricyclic antidepressants have been empirically used for years to treat psychic aspects in patients with

bronchial asthma. They may however improve pulmonary function by at least three different pharmacol-

ogical mechanisms. We tested amitriptyline in unanaesthesised, ovalbumine - sensitised guinea pigs in

the two-chamber whole-body plethysmograph by Dorsch and Hess. The results were compared to the

selective serotonin reuptake inhibitor fluoxetine. The variable “compressed air” (CA) over time (t) de-

scribes the degree of bronchial obstruction. Amitriptyline, applied 60 minutes before ovalbumine chal-

lenge (10 mg/kg) significantly reduced bronchial obstruction. Mean AUC of the CA/t-function was 0.96

[ml*min] in 15 amitriptyline treated animals as compared to 8.99 [ml*min] in 14 placebo treated animals

(p<0,05). Amitriptyline, once a day for 4 weeks (2 mg/kg / 10mg/kg) did not prevent bronchial obstruction.

Fluoxetine was ineffective. The short term effect of amitriptyline on allergen induced bronchial obstruction

is clear evidence for a direct bronchodilating action of tricyclic antidepressants.

K. Haerter, A. Vroon, A. Kavelaars, V. Limmroth, C.J. Heijnen, M. Schedlowski, S. Elsenbruch

Department of Medical Psychology and Clinic of Neurology, University of Essen; Department of Pediatric

Immunology, University Medical Center, Utrecht

29. Adrenergic modulation of cytokine production during MOG-induced EAE

In vivo experiments have shown adrenergic modulation of experimental autoimmune encephalomyelitis

(EAE), a rat model of multiple sclerosis. The goal of this study was to investigate mechanisms of adren-

ergic modulation in EAE using in-vitro techniques. Therefore, the influence of the beta2-agonist terbuta-

line on LPS-stimulated TNF production by peritoneal macrophages and the effects of the alpha-1 agonist

methoxamine and the alpha-2 agonist UK-14304 on ConA-stimulated splenocyte proliferation and cyto-

kine production were measured during various stages of the disease compared to controls. The results

showed significant disease stage-dependent alterations in TNF production by LPS stimulated macro-

phages, with maximal production during remission (p<0.01 compared to controls). Terbutaline inhibited

macrophage TNF production in a dose-dependent manner at concentrations between 1 and 1000 nM with

nearly complete suppression at 300 nM (83.75 ± 3.8% decrease from 0 nM, p<.001). However, there

were no differences between EAE and controls in TNF inhibition with terbutaline. Splenocyte proliferation

was slightly increased with both alpha agonists in all groups. Splenocyte production of TNF was lowest in

second peak animals (542.9 ± 52 pg/ml vs. 581.3 ± 80 first peak vs. 704.1 ± 78 controls). Splenocyte IFN-

gamma production was highest in EAE second peak (3443.8 ± 1194 pg/ml) compared to EAE first peak

(2156 ± 515, p<.05) and controls (2711.4 ± 778, p<.1). However, there was no effect of either alpha-

agonist on TNF or IFN-gamma production in controls or EAE animals. Together, these results support

that the disease stages of EAE are associated with specific changes in the capacity of splenocytes and

peritoneal macrophages to produce cytokines. Beta-2 adrenergic inhibition of TNF production by perito-

neal macrophages does not differentiate EAE from controls. Alpha adrenergic substances enhance

splenocytes proliferation in EAE and controls, but have no effect on splenocyte TNF or IFN-gamma

production in EAE or controls.

P. Hauck, U. Ungethüm, A. Schiffold, C. G. O. Baerwald, M. Wahle, and A. Krause

Department of Internal Medicine III, Charité, Berlin, Germany; and Department of Internal Medicine IV,

University Leipzig, Germany

30. The suppressive effect of acetylcholine on monocyte activation is mediated by nicotinergic receptors

The immune response is under partial control of the autonomic nervous system. However, only little is

known about the direct effect of acetylcholine (ACh) on the function of immune cells. The study aimed at

investigating the impact of ACh on monocyte activation and the expression of ACh receptors (AChR) on

monocytes. Mononuclear cells from 24 healthy donors were stimulated with LPS (1µg/ml) and co-

incubated with ACh in final concentrations between 10(-5) and 10(-13) mol/l. After 24 hours, IL-6 as a

marker of monocyte activation was determined in culture supernatants. The results showed that in a sub-

group of 5 subjects ACh had a strong dose-dependent inhibitory effect on monocytes activation with a

decrease of IL-6 concentrations by 70% (p<0.001). In another subgroup (n=5) ACh only had moderate

inhibitory effects (maximum IL-6 decrease 35%) while monocytes from the remaining 14 subjects did not

respond to ACh at all. ACh effects in the responder groups could be blocked by hexamethonium but not

by atropine, indicating that ACh effects were mediated via nicotinergic AChR (nAChR). Moreover, it could

be shown that these nAChR did not bind alpha bungarotoxin (bgt) and therefore belong to a special sub-

group of non-bgt binding nAChR. Using flow cytometry and molecularbiological methods it could be dem-

onstrated that monocytes indeed express nAChR consisting of alpha 3, alpha 5 and beta 2 subunits. So

far, these nAChR have only been found on ganglionic postsynaptic membranes and keratinocytes. Stud-

ies on the regulation of AChR expression on immune cells and the impact of ACh on the immune re-

sponse under physiologic and pathologic conditions are currently under way.

C.J. Heijnen1, M.S. Lombardi1, A.Vrooni1, R. Jacobs2, R.E. Schmidt2, M. Schedlowski3, A.Kavelaars1

1Laboratory for Psychoneuroimmunology, University Medical Center Utrecht, Utrecht, the Netherlands

2Department of Clinical Immunology, Hannover Medical School, Germany 3Department of Medical Psy-

chology, University of Essen, Germany

31. G protein coupled receptors and neuroendocrine signalling in autoimmunity

G protein coupled receptor kinases (GRK) play a key role in regulating desensitisation of G protein cou-

pled receptors such as ß2-adrenergic, SP, opioid and chemokine receptors. High levels of GRK are pre-

sent in the immune system. We recently reported changes in GRK levels in peripheral blood mononuclear

cells of patients with rheumatoid arthritis. The reduced GRK level was associated with increased reactivity

of PBMC to ß- adrenergic agonists. The aim of this study was to investigate whether changes in GRK

levels in the immune system and in non-immune organs occur during the course of inflammatory autoim-

mune diseases. In a rat model of adjuvant- induced arthritis (AA), we investigated GRK activity and ex-

pression in primary and secondary lymphoid organs and in heart and pituitary. At the peak of the inflam-

matory process, we observed a profound down regulation of GRK-2, GRK-6, and GRK-3 in splenocytes

and mesenteric lymph nodes cells from AA rats. In contrast, no changes in GRK were observed in a pri-

mary lymphoid organ or in non-immune organs such as heart and pituitary. During the remission phase of

AA GRK levels in spleen and mesenteric lymph nodes return to baseline levels. To answer the question

whether reduced GRK levels in immune organs also occur during other inflammatory diseases, we also

analysed cells from animals with experimental autoimmune encephalomyelitis. Finally, using a GRK2

heterozygous knock out mouse, we show that reduced GRK2 levels influence the course of experimental

autoimmune encephalomyelitis as well as the response to chemokines.

J. Herklotz, J. Westermann

Department of Anatomy (4120), Hannover Medical School, Hannover, Germany; Institute of Anatomy,

Medical University of Lübeck, Lübeck, Germany

32. Local immune response in the spleen: Regulation by the nervous system and effects on behaviour

We investigated whether a local immune response is regulated by the nervous system and whether it

induces changes in the behaviour of rats. A local immune response in the spleen was induced by intrave-

nous injection of sheep red blood cells (SRBC) into Lewis rats. After 10 days the size of the different

splenic compartments, the rate of proliferation and apoptosis, and the formation of germinal centres was

analysed. The behaviour was investigated by recording home cage activity continuously for 10 days. The

role of the nervous system was studied by performing surgical denervation of the spleen 5 days before

induction of the immune response. Denervation reduced norepinephrine content to less than 10% of the

normal level, but did not affect compartment size, proliferation and apoptosis. Plasma interleukin-2 levels

were elevated, whereas IL-4, IL-6, IL-10 and IFN-g remained normal. The immune response led to a sig-

nificant increase in the number and size of germinal centres in the intact spleen which was not influenced

by denervation. Immediately after injection of SRBC the activity of the rats with intact spleens decreased

compared to saline injected animals. Thereafter, no differences were seen between both groups, and

denervation did not change this pattern. Our study demonstrates that an intact innervation of the spleen is

not necessary for the induction of a local immune response. In addition, the local immune response af-

fects neither via afferent innervation nor via secreted cytokines the behaviour as measured by the un-

changed home cage activity. Thus, the presence of molecules and its respective receptors as such is not

sufficient to predict an interaction between the immune and the nervous system.

H. Hutzelmeyer, D. v. Holst


33. The critical first year in the life of European wild rabbits (Oryctolagus cu-niculus)

Wild European rabbits live in groups of 1-3 adult males and 1-6 adult females; both sexes have separate

linear rank orders, which are established and maintained by intensive fights. Data are presented of a

study on population physiology and behavioural ecology in wild rabbits living in a 22,000 m² enclosure.

This report focuses on the relevance of behaviourally induced immunomodulation for the reproductive

success of females. Since 1987, a yearly average of 36.3+3.2 adult females with their offspring lived in 8-

14 territorial groups in the enclosure. The reproductive season lasted from March to September. Since

female rabbits exhibit postpartum oestrus they potentially could give birth to up to 7 litters at monthly in-

tervals. However, the number of reproducing females decreased during the reproductive season from

about 80% to less than 10% reducing the mean number of litters per female and year to 3.18+0.05

(n=497) with a mean litter size of 4.83+0.04 offspring (n=1575). From the total of 7608 kittens only

5.5+1.3% survived to adulthood (= breeding season the year following their birth). Predation as well as

intestinal diseases (caused by several species of Eimeria and nematodes) represented the major mortal-

ity factors. The reproductive success of females was influenced to a large extent by their social ranks.

This depended on two effects of about equal strength: a higher fecundity of high ranking females and a

lower mortality of their offspring between birth and adulthood. The relevance of behavioural induced im-

munological changes for these differences in offspring mortality is shown. Supported by the Volkswagen

Foundation (I/75182).

R. Jacobs1, C.R. Pawlak2, E. Mikeska2, D. Meyer-Olsoni1, M. Martin3, C.J.Heijnen4, M.Schedlowski5,

R.E. Schmidt1

1Divisions of Clinical Immunology, 2Medical Psychology, 3Clinical Molecular Pharmacology, Hannover

Medical School, 30623 Hannover, Germany. 4Division of Immunology, University Hospital for Children

and Youth, Utrecht, The Netherlands, 5Division of Medical Psychology, University of Essen, Essen, Ger-

many

34. SLE and RA patients differ from healthy controls in their cytokine pattern after stress exposure

To study whether patients with rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE) differ

from healthy individuals in their immune responses under acute psychological stress. Methods. Numbers,

phenotype, and function (NK cytotoxicity and cytokine production) of peripheral blood mononuclear cells

(PBMC) were analysed at three time points before and after stress exposure in female patients and age-

matched healthy subjects. Acute psychological stress was induced by a public talking task. All patients

displayed mild disease activity and were all free of immunosuppressive medication. Results. Natural killer

(NK) cell numbers increased transiently in all groups under stress. NK activity, however, increased in

healthy controls (HC) only. We observed a stress induced increase in interleukin (IL)-4 producing (IL-4+)

cells in SLE patients only, whereas interferon (IFN) gamma+ cell numbers increased due to stress in all

three groups as assessed by FACS from intracellular staining of PMA/ionomycin stimulated PBMC.

Analysis of supernatants from whole blood PHA cultures revealed increased IFN gamma and IL-10 levels

in healthy subjects but neither in SLE nor in RA patients after stress exposure. Conclusions. These data

demonstrate that RA and SLE patients differ in their immune response under stress from healthy controls.

Changes in cytokine patterns might contribute to stress-induced exacerbation of disease activity in RA

and SLE patients. supported by the Volkswagen Foundation Germany, Grant I/72 032/031

R. Joachim, D. Quarcoo1, A. Zenclussen, J. Niess, P. Arck and B.F. Klapp

Department of Internal Medicine and 1Department of Pediatrics, Charité, Humboldt University, Berlin,

Germany

35. Stress-induced increment on bronchoalveolar cell count in OVA- sensitised mice is mediated by tachykinin NK1 receptor

Airway inflammation is a leading symptom of bronchial asthma. Recent data is linking stress to the initia-

tion and perpetuation of asthma symptoms. Tachykinins like substance P have been shown to play an

important role in generating asthma symptoms and furthermore to be released in tissues in response to

stress. The aim of the study was to determine 1) the influence of stress on murine allergen induced air-

way inflammation and 2) the involvement of tachykinins in stress mediation of airway inflammation. CBA/J

mice were sensitised by i.p. injection of ovalbumine (OVA). Coinciding with the beginning of the first OVA

aerosol airway provocation, one half of the mice was stressed by exposure to an ultrasonic sound

stressor for 24 hours. Part of the animals were additionally treated with a tachykinin NK1-receptor an-

tagonist (NK1-RA) before airway challenges. 24 h after the second airway provocation, animals were

sacrificed. Broncho-alveolar lavage fluid (BAL) was obtained and total cell numbers were determined.

Cytospins were prepared for each sample and after staining differential cell counts were performed. A

significantly higher total cell count was present in the BAL in stressed animals compared to unstressed

control mice. This stress-induced increase was mainly caused by an increase of eosinophils and could be

blocked by application of the NK1-RA, whereas NK1-RA did not influence airway inflammation in un-

stressed controls. Our data suggest that stress increases allergen induced airway inflammation. Further-

more it points to tachykinins like substance P as putative mediators in stress induced airway inflamma-

tion.

K G Kahl1,2, N Kruse3, M H Schmidt1, P Rieckmann3

1 Department of Child and Adolescent Psychiatry and Psychotherapy, Central Institute of Mental Health,

Mannheim; 2Department of Psychiatry and Psychotherapy, Medical University of Lübeck; 3Department of

Neurology, Clinical Research Unit for Multiple Sclerosis and Neuroimmunology, Julius-Maximilians Uni-

versity, Würzburg

36. Quantitative analysis of cytokine mRNA expression patterns in the disease course of adolescent patients with anorexia nervosa

Anorexia nervosa (AN) is a serious eating disorder characterised by extreme weight loss and abnormali-

ties of the neuroendocrine and immune systems. Cytokines have been discussed to be involved in the

pathomechanisms underlying cachexia, although different results were obtained in anorexia nervosa by

detecting cytokines on the protein level. Therefore our study aimed at examining the mRNA expression

pattern of tumor necrosis factor-alpha (TNF) , interferon-gamma (IFN-gamma), interleukin-6 (IL-6) and

interleukin-10 (IL-10) in these patients. A sensitive quantitative polymerase chain reaction (PCR) method

was applied to determine cytokine mRNA levels in patients with anorexia nervosa (n=11) and age and

sex matched control subjects (n=10) in a whole blood assay. We found a significant increase in TNF and

IL-6 mRNA expression in anorectic patients at admission (mean BMI 14.8 ± 1,3) compared to control

subjects. No significant group differences were found concerning the expression of IFN-gamma and IL-10

mRNA expression at admission. During follow-up, the expression of TNF mRNA remained high in anorec-

tic patients (mean BMI 18.7 ± 0.5) while IL-6 mRNA expression decreased to the range of control sub-

jects. We conclude that quantitative real time PCR may be a useful tool to detect differences in the mRNA

expression pattern of cytokines. Our results suggest that TNF may contribute to metabolic abnormalities

in anorexia nervosa even after goal BMI is achieved.

T. Karl1, K. Shingu1, R. Jacobs2, R.E. Schmidt2, R. Mentlein3, S. Kaiser4, N. Sachser4, R. Pabst1, St.

von Hörsten1

1Department of Functional and Applied Anatomy, Medical School of Hannover, Germany, 2Department of

Clinical Immunology, Medical School of Hannover, Germany, 3Departemnt of Anatomy and Cell Biology;

University of Kiel, Germany, 4Department of Behavioural Biology, University of Münster, Germany

37. CD26 mutant rats: A model for psychoneuroimmune interactions

The peptidase CD26 is specifically involved in truncation of circulating neuropeptides (NPY), hormones

(GLP-1, GHRH) and chemokines (eotaxin, RANTES), in immune responses and cell adhesion. Possibly,

CD26 activity alters behavioural, neuroendocrine and immune regulation at various levels. In the present

studies, mutant Fischer 344 (F344) substrains that lack endogenous CD26-like enzymatic activity and/or

expression (F344GER and F344JAP substrains) were compared with wild-type F344USA rats. F344 rats

of each substrain were characterised in a behavioural tests battery, in expression studies, in tests of glu-

cose tolerance as well as neuroendocrine stress responsiveness, and in immunoassays for NK cell func-

tion, tumor adhesion, and colonisation. Monitoring growth, neurological, and motor functions revealed no

overt differences although differential mutations in F344GER and F344JAP were confirmed. Mutant rats

showed a improved glucose tolerance and a higher food intake but a reduced water intake suggesting

functionally relevant differences in GLP-1 and NPY metabolism. These differences may also account for a

reduced behavioural and neuroendocrine stress response in these mutant F344 substrain. Furthermore,

NK cell function and metastasis were altered depending on the type of mutation. Overall, these data indi-

cate that altered expression and function of a single enzyme the CD26 protein can drastically change the

outcome of behavioural, neuroendocrine and immunological responses. Mutant F344 substrains repre-

sent a valuable animal model for pathophysiological changes related to a differential function and/or ex-

pression of CD26. Supported by the DFG/GK705 (TK) and the Volkswagen Foundation /I/75169 (RP and

SvH).

S. Kastenbauer, U. Ködel, T. Brzoska1, TA. Luger1, HW Pfister

Department of Neurology, Clinic Großhadern, Ludwig-Maximilians University, Munich, and 1Department

of Dermatology, University of Munster, Germany

38. Failure of alpha-melanocyte stimulating hormone to attenuate cerebral com-plications in experimental pneumococcal meningitis

Alpha-melanocyte-stimulating hormone (alpha-MSH) is an endogenous neuroimmunomodulatory peptide

that can inhibit a broad range of inflammatory mediators known to be involved in the pathophysiology of

bacterial meningitis. We evaluated the effect of alpha-MSH in a rat model of pneumococcal meningitis.

Six hours after intracisternal infection with Streptococcus pneumonia, rats were left untreated (n = 10) or

treated with a single intraperitoneal injection of 100 µg/kg alpha-MSH (n = 9), 100 µg/kg intraperitoneal

alpha-MSH every four hours (n = 10) or 100 µg/kg intracisternal alpha-MSH every four hours (n = 3).

Twenty-four hours after infection, the intracranial pressure and CSF bacterial titres were determined.

Blood-brain barrier disruption was evaluated by fluorescence microscopic detection of Evans Blue in brain

slices. In brain homogenates, cytokine concentrations were measured by ELISA and the leukocyte accu-

mulation was determined by myeloperoxidase-assay. CSF alpha-MSH concentrations were quantified by

RIA. Both systemic and intracisternal alpha-MSH failed to influence blood-brain barrier disruption, in-

creased intracranial pressure, brain cytokine concentrations (IL-1 beta, IL-6, TNF, MIP-2, and IL-10), CSF

bacterial titres, and clinical parameters of disease severity (weight loss, body temperature, and blood

pressure), although the treatment strongly increased the CNS concentrations of alpha-MSH. However,

systemic but not intracisternal alpha-MSH slightly reduced the CNS leukocyte accumulation, indicating

that the leukocyte extravasation is inhibited by alpha-MSH from the blood side. Our results show that

alpha-MSH reduces the CNS leukocyte accumulation by its systemic action, but does not attenuate men-

ingitis-associated intracranial complications.

S. Kerzel1, G. Päth1, D. Quarcoo1, U. Raap1, H. Renz1 and A. Braun2

1Department of Clinical Chemistry and molecular Diagnostics, Philipps University Clinic, Marburg,

2Fraunhofer Institute for Toxikology und Aerosol Research, Pharmaceutical Research, and Clinical Inha-

lation, Department of Immunology/Allergology, Hannover

39. Neuronal plasticity is mediated by neurotrophin receptor p75NTR in a mur-ine model of allergic bronchial asthma

A key feature of chronic asthma is airway hyperresponsiveness to unspecific stimuli (e. g. smoke). This

might be due to enhanced neuronal reflex circuits controlling airway function. The underlying neuronal

plasticity comprises lowering of the firing threshold of sensory airway nerves and increased synthesis of

proinflammatory neuropeptides (tachykinins like substance P). There is growing evidence that nerve

growth factor (NGF), released by invaded immune cells, is responsible for this effect of allergic inflamma-

tion. To asses the mechanisms of NGF action, the role of its low-affinity neurotrophin receptor p75NTR

was examined in a murine model of allergic asthma. p75NTR -/- mice as well as anti-p75NTR-antibody

treated wildtype animals were sensitised to ovalbumine. Allergic airway inflammation was induced by

aerosolic allergen challenges. A characteristic feature of allergic wildtype mice is enhanced neuronal re-

sponse to inhaled capsaicin, assessed by head-out body plethysmography as lengthening of phase I of

expiration. Blocking of p75NTR function prevented this augmented reflex activity almost completely. In

addition, allergic inflammation was markedly decreased, characterised by the lowered influx of eosino-

phils into the airways. The receptor distribution in wildtype animals and the diminished substance P con-

tent in lungs of p75NTR -/- mice suggest that these effects are partly mediated via sensory nerves and

are possibly due to neurogenic inflammation. Taken together, p75NTR might be a key player in inflamma-

tion-induced neuronal plasticity.

B.C. Kieseier1, A. Chan2, R. Gold2, K.V. Toyka2 and H.P. Hartung1

1Department of Neurology, Karl-Franzens-University, Graz, Austria, 2Department of Neurology, Julius-

Maximilians-University, Würzburg, Germany

40. Expression of TNF-converting enzyme in inflammatory demyelination of the peripheral nervous system

Tumor necrosis factor-alpha (TNF) is a major proinflammatory cytokine implicated in the pathogenesis of

inflammatory demyelinating diseases of the peripheral nervous system (PNS), such as the Guillain-Barré

syndrome (GBS). Soluble TNF is released from its membrane-bound precursor by shedding through a

proteinase, identified as TNF-converting enzyme (TACE; ADAM-17), a member of the ADAM (a disin-

tegrin and metalloproteinase) domain family of proteins. To study the role of this proteinase in inflamma-

tory demyelinating disorders of the PNS the expression and distribution patterns of ADAM-17 were inves-

tigated immunohistochemically in adoptive transfer experimental autoimmune neuritis (AT-EAN), an ex-

perimental model of the GBS. Immunoreactivity against ADAM-17 could be found in diseased animals

with peak expression concurrent with maximal disease severity. Double immunofluorescence labelling

pointed to T cells as the cellular source of this enzyme. These data were strengthened by studying sural

nerve biopsies from GBS patients. Positive immunoreactivity against ADAM-17 could be found in the

biopsies. Moreover, increased protein expression was detectable in the cerebrospinal fluid of GBS pa-

tients in comparison with non-inflammatory controls. Our findings suggest that ADAM-17 is upregulated

during AT-EAN and is expressed in the GBS and thus may contribute to the pathogenesis of inflammatory

disorders of the peripheral nervous system.

C. Kirschbaum, L. Joksimovic, N. Rohleder, J. Wolf, S. Riegler

Department of Experimental Psychology, University of Düsseldorf, Düsseldorf, Germany

41. Posttraumatic stress disorder and HPA functioning: Hypocortisolism and increased glucocorticoid sensitivity of PBMC in Bosnian war victims

Posttraumatic stress disorder (PTSD) has been repeatedly found to be associated with a state of hy-

pocortisolism as characterised by a lowered circadian secretion of cortisol. Subsequent studies revealed

that these PTSD victims have an increased number of glucocorticoid (GC) receptors in peripheral blood

mononuclear cells (PBMC). The functional significance of these alterations remained elusive. Thirteen

Bosnian war PTSD victims and 15 age and sex matched control subjects were studied. On two consecu-

tive days, they collected saliva samples during the first hour of awakening and at 11, 15, and 20 hrs, re-

spectively. In addition, GC sensitivity was measured by DEX-inhibition of LPS-induced interleukin-6 (IL-6)

and tumor necrosis factor alpha (TNF) production in whole blood. Results showed that the PTSD group

had lower salivary cortisol levels throughout the day (F=9.07, P=0.006). PTSD subjects did not show the

expected cortisol response after awakening and their daytime cortisol levels remained significantly lower

than in the control group. Interestingly, the cortisol levels upon awakening and at 20 hrs did not differ

between groups. Concomitantly, a much lower dose of DEX was required for IL-6 and TNF suppression

(t=2.86, P=0.009; TNF: t=4.42, P<0.001) reflecting a higher GC sensitivity of PBMC in PTSD victims.

These data show that in war victims suffering from PTSD, hypocortisolism is associated with an increased

GC sensitivity. Whether this pattern reflects an adaptive mechanism or might render these individuals

more vulnerable to certain diseases in which inflammation plays a critical role for the host defence re-

mains to be elucidated.

J.M. Kittner1, R. Jacobs1, C.R. Pawlak2, C.J. Heijnen3, M. Schedlowski4, R.E. Schmidt1

1Department of Clinical Immunology, Hannover Medical School, Germany, 2Department of Physiological

Psychology, University of Marburg, Germany, 3Wilhelmina Childrens Hospital,Utrecht, the Netherlands,

4Department of Medical Psychology, University of Essen, Germany

42. Effects of adrenaline infusion on immune parameters in rheumatoid arthritis (RA) patients and healthy controls (HC)

A dysregulation of the adrenergic system is suggested to affect disease activity in RA. Aim of this study

was to investigate whether the immune response under adrenaline is different in RA patients. 15 female

RA patients and 14 healthy controls were infused with 1 ug/kg adrenaline over 20 min. Blood was drawn

before, immediately after and one hour after end of the infusion. Phenotype of leukocytes and function

like cytokine production and cytotoxicity were determined by FACS or 51Cr release assay, respectively.

All subjects exhibited mild cardiovascular changes with an increase of heart rate (20%), an increase of

the systolic (5%), and a decrease (15%) of the diastolic blood pressure. NK cells increased by factor 5.7,

CD3+cells by 1.5, monocytes by 1.6, and neutrophil granulocytes by 1.2 in both groups. Preferentially,

activated monocytes (CD14+CD16+) and T cells perforin+CD3+, CD11a++CD8+) were recruited in both

RA and HC. IL-2, IL-4, IFN-gamma, and TNF producing lymphocytes were increased after infusion with-

out differences between patients and controls. IL-8 and IL-10 producing monocytes were higher in pa-

tients and presented a significant higher increase after the infusion compared to HC. Cytotoxic activity

was higher in RA patients and exhibited an increase after the infusion. All values returned to baseline one

hour later. Adrenaline infusion is a valid model to mimic acute psychological stress. Our data demonstrate

an altered response to adrenaline in patients with RA with both pro- and anti-inflammatory dissimilarities

compared to HC. Activated T-cells and monocytes recruited by adrenaline may influence disease activity.

Supported by grant I/72 032/031 of the Volkswagen Foundation

C. Körner1, B. Wieland1, W. Richter2, H.-M. Meinck1

Departments of 1Neurology and 2Orthopedic Surgery, Ruprecht-Karls-University, Heidelberg, Germany

43. Stiff man-syndrome, a neuropsychiatric disorder with autoimmunity against GABAergic neurones. Part II: GAD autoantibodies impair GABAergic func-tions within the motor cortex

In the stiff man syndrome (SMS) and its variants, dominant autoimmunity is targeted against glutamic

acid decarboxylase (GAD-Ab), the rate-limiting enzyme of GABA synthesis. We have recently shown that

GAD-Ab effectively inhibit GABA synthesis in vitro. This and various other observations suggest that ex-

cess alpha motoneurone activity in SMS and its variants might be caused by attenuation of GABAergic

inhibition. In the human motor cortex both, intrinsic inhibition (ICI) and facilitation (ICF) are thought to be

mediated by GABAergic mechanisms. We investigated ICI and ICF with transcranial magnetic stimulation

(TMS, double shock paradigm) in a total of 21 patients with SMS (n=15) or PERM (n=6) and in 14 healthy

controls. Seven patients received no GABAergic medication. Serum levels of GAD-Ab were determined

by radioimmunoassay. ICF was significantly enhanced, and ICI reduced in patients with SMS and PERM.

In patients treated with GABAmimetic drugs, this effect was partially masked. Reduced ICI and enhanced

ICF of the motor cortex were positively correlated with elevated GAD-Ab levels. However, reduced intrin-

sic GABAergic inhibition in the motor cortex was not associated with clinical signs of motor cortex hyper-

excitability, nor with reduced motor threshold on TMS, enlarged somatosensory evoked potentials or

transcortical long-loop reflexes. Our results demonstrate that defective GABAergic intrinsic inhibition in

the motor cortex of patients with SMS and PERM is correlated with elevated levels of GAD-Ab suggesting

that autoimmunity against GAD plays a causative role. However, the observed physiological dysfunction

is subclinical and does, moreover, not lead to a net hyperexcitability of the motor cortex as revealed by

established physiological tests.

D. Köthe, M. Haack, D. Hinze-Selch, Th. Kraus, A. Schuld, Th. Pollmächer

Max Planck Institute of Psychiatry, Munich, Germany

44. Body weight and the plasma levels of leptin and endogenous immune modu-lators during treatment with carbamazepine

Several antipsychotic agents induce weight gain and increase circulating levels of cytokines. To asses

whether carbamazepine have similar effects we measured weekly plasma cytokine levels and weight in

12 inpatients, suffering of various psychiatric disorders during the first four weeks of treatment. Treatment

with carbamazepine lead to a significant increase of weight and the BMI (p=0,044; p=0,046) across five

weeks, preceded by a significant increase of TNF plasma-levels in the first two weeks of treatment.

Plasma levels of the soluble TNF-receptors p55 and p75, sIL2R, IL6, and circulating levels of leptin were

not affected. We conclude that weight gain induced by psychotropic agents may occur without increased

circulating levels of leptin, but there is a possibility that the activation of the TNF system might be an early

and sensitive marker of ensuing weight gain.

C Kohl1, T Walch1, R Huber1, B Widner4, A Laich4, E Sölder3, J Kemmler1, H Schröcksnadel3, D

Fuchs4, B Sperner-Unterweger2

1Department of Psychiatry, 2Department of Biological Psychiatry, 3Department of Gynaecology,

4Institute of Medical Chemistry and Biochemistry, University of Innsbruck, Austria

45. Changes of tryptophan and kynurenine in the postpartum period

Postpartum blues is a mild form of postpartum mood disorders. It is associated to biochemical changes

during and in the first days after birth. One of these alterations is an increase of serum tryptophan (TRP),

an amino acid, that is - according to animal studies - involved in the process, which prevents fetal rejec-

tion and plays a role in depressive disorders. Our study was designed to examine alterations of trypto-

phan and its catabolic product kynurenine in the postpartum period and to compare these changes in

mothers with and without postpartum blues. 95 healthy women delivering without complications provided

blood during parturition, 2 and 4 days after birth. The blood samples were analysed by HPLC (high pres-

sure liquid chromatography). Women were asked to perform the Edinburgh Postnatal Depression Scale

(EPDS) on day 2 and 4. Eight point six percent (n=9) of the women met the criteria of postpartum blues

according to EPD scale (group2). In women without blues (group 1; n=86) tryptophan increased after birth

to reach normal levels on day 2, whereas group 2 did not show this increase until day 4. The difference in

the change between day 0 and 2 reached statistical significance (p=0.027). Kynurenine increased be-

tween day 0 and 2 in group 1, but not in group 2 (p=0,012). Our study shows a difference in the altera-

tions of tryptophan in women after parturition with and without postpartum blues, and also – surprisingly –

a difference in the level of kynurenine.

J. Kraus, C. Börner, E. Giannini, K. Hickfang, V. Höllt

Department of Pharmacology and Toxicology, University of Magdeburg, Magdeburg, Germany

46. Mu opioid receptor expression in immune cells is induced by IL-4 and TNF

The aim of this study was to investigate regulation of mu opioid receptor expression in immune cells by

cytokines. Under unstimulated conditions no mu receptor transcripts were found by RT PCR in immune

cells (cell lines of B cells (Raji), monocytes (U937) and endothelium (HMEC-1); primary T cells and

granulocytes). A dramatic induction of transcription was observed in all these cells after stimulation with

either IL-4 or TNF. Transient expression of reporter gene constructs in combination with promoter dele-

tion and mutation analysis in Raji, HMEC-1 and SH SY5Y cells revealed a single cis-active promoter ele-

ment in the human gene at nt -997 and the rat gene at nt -727 conferring IL-4 regulation of mu transcrip-

tion. Gel shift assays (EMSA) identified the elements as binding sites for transcription factor Stat-6. Fur-

thermore, reporter gene analysis and EMSA revealed that TNF regulates mu receptor transcription via

NFkappaB, for which several binding sites on the gene promoters were found. Interestingly, a polymor-

phism has been reported in humans within the Stat-6 binding site which decreases transcription factor

binding and trans-activation of reporter gene transcription.

H. Krönig, M. Riedel, M. J. Schwarz, M. Ackenheil, N. Müller

Department of Psychiatry, Ludwig-Maximilian University Munich, Nussbaumstr.7, 80336 Munich, Ger-

many

47. Genetic investigations based on immunological findings in schizophrenia

Immunological studies have described higher IL-6 levels in serum of schizophrenic patients (1), lowered

IL-2 in-vitro production in schizophrenia (2), lower levels of the cell adhesion molecule ICAM-1 (3) and a

significant increase of IL-10 in chronically ill schizophrenic patients (4). As a genetic component has been

suggested in the aetiology of schizophrenia, we investigated functional polymorphisms in the genes of the

above described immune parameters: G174C polymorphism in the IL-6 gene promotor; T330G polymor-

phism in the IL-2 promotor; G469A polymorphism in the exon 6 of the ICAM-1 gene; G 1082A promotor

polymorphism in the IL-10 gene. 127 schizophrenic patients and 133 healthy control individuals were

included. PCR was carried out as previously described. Clinical variables were monitored at the time of

admittance and before discharge from the hospital by use of the PANSS. No significant difference be-

tween patients and controls was found in allele frequencies of the IL-2, IL-6 and ICAM-1 polymorphisms.

However there was a significant increase of the homozygotic A-Allele between patients and controls for

the IL-10 polymorphism, that is shown to be associated with decreased IL-10 production. Moreover there

was a marked relationship between the homozygosity for the A allele and the more pronounced therapy

response regarding the positive and global symptomatology. Altogether this could demonstrate an asso-

ciation between immunogenetic markers and clinical variables of therapy response.

(1) Lin A et al. (1998) Schizophr Res 32:9-15

(2) Arolt V et al.(2000) Mol Psychiatry 5:150-158 (3) Schwarz MJ et al. (2000) Biol Psychiat 47:29-33 (4) Cazullo C.L. et al. (1998) Prog.Neur.Biol.Psychiatry 22:947-957

H. Krönig, M. J. Schwarz, M. Riedel, M. Ulmschneider, R. Yolken1, N. Müller

Psychiatric and Psychotherapeutic Hospital, Ludwig-Maximilian University Munich, Munich, Germany;

1Stanley Division of Developmental Neurovirology, Johns Hopkins Univ., School of Medicine, Baltimore

MD, USA

48. TNF polymorphism G308A may protect from cytomegalovirus infection in schizophrenia

The TNF G308A polymorphism is described to be associated with higher TNF production

compared to the wild type. Moreover, it has been shown that the A-allele is also associated

with lower cytomegalovirus antibody titres in healthy blood donors. We analysed 35 schizo-

phrenic patients with regard to the TNF G308A polymorphism and the anti-CMV antibody-

titres. We found a statistically significant difference (p < 0,002) regarding the anti-CMV titres

between patients with the A-allele (n = 10) and the G-allele (n = 25). A-allele positive patients

had significantly lower anti-CMV titres. This result supports the literature and implicates that

the A-allele might have a protective function for CMV infection. Moreover, the TNF polymor-

phism may help to define subgroups of schizophrenics in which CMV infection may play a role,

or not. However, statistical limitations have to be discussed. Supported by the Vada and

Theodore Stanley Foundation.

S R. Kunz, A. Steptoe, C. Kirschbaum, M. Marmot

Department of Epidemiology and Public Health, University College London, London, United Kingdom

49. Differential effects of psychological stress on IL-6 and IL-1ra responses in women and men from the Whitehall II cohort

One important mechanism of glucocorticoids is to downregulate immune activity to prevent host destruc-

tiveness after disease. Heightened levels of proinflammatory cytokines such as IL-6 and IL-1 have been

found in patients with cardiovascular diseases, atherosclerosis and depression. We investigated varia-

tions of endocrine and cardiovascular responses and their influence on proinflammatory cytokines as well

as at associations with disease symptoms. All 149 participants were a healthy subset of the Whitehall II

cohort. Saliva samples were measured five times throughout the session and blood was taken after a 30

min rest period, immediately after two stress tasks (Stroop colour word test, mirror tracing) and after a 45

min recovery period. We looked at cortisol responses following the tasks. 79 individuals (Responders)

showed a strong cortisol response following these tasks, while 70 other participants (Non-Responders)

showed a blunted response. Cortisol-Non-Responders showed a tendency for a higher heart rate and a

lower heart rate variability during baseline and recovery than the Cortisol- Responders. No group differ-

ences concerning cardiovascular measures were found for the stress period. Both groups showed an

increase in IL-6 levels after stress, but the Cortisol-Non-Responders only showed an increase of IL-1ra

after stress whereas Cortisol-Responders showed a decrease. An increase of IL-1ra was also positively

associated with depressive symptoms. We conclude that individual variations in neuroendocrine stress

responsivity may have different impact on proinflammatory cytokines. A higher sympathetic activity com-

bined with a blunted cortisol response could result in increased levels of IL-1ra and may in turn be rele-

vant to health risk.

T. Lange, B. Perras, H.L. Fehm, J. Born

Clinical Research Group Neuroendocrinology and Department of Internal Medicine, University of Lübeck,

Germany

50. Sleep enhances the human antibody response to hepatitis A vaccination

The common belief that sleep supports immune defence has received surprisingly little direct experimen-

tal support. The antibody response to vaccination provides a tool to assess the influence of sleep on

adaptive immune functioning in humans, which is valid and clinically relevant. Two groups of healthy hu-

mans (n=19) not previously infected with hepatitis A virus (HAV) were studied. On the night following

primary vaccination with inactivated HAV which took place at 0900 h, one group had regular sleep. The

other group stayed awake, and did not sleep before 2100 h the following day. HAV antibody titres were

measured repeatedly until 28 days after vaccination. Plasma hormone concentrations and white blood

cell (WBC) subset counts were determined on the night and day after vaccination. Subjects who had

regular sleep following vaccination, displayed after four weeks a nearly twofold higher HAV antibody titre

than subjects who stayed awake on this night (p=0.018). Compared with wakefulness, sleep after vacci-

nation distinctly increased release of several immune stimulating hormones including growth hormone,

prolactin and dopamine (p<0.01). Concentrations of thyrotropin, norepinephrine and epinephrine were

lowered by sleep (p<0.02), while sleep only marginally influenced WBC subset counts. Our data suggest

that nocturnal sleep improves the formation of immunological memories as reflected by antibody produc-

tion in humans. Sleep presumably acts by inducing a hormonal environment in lymphoid tissues enhanc-

ing lymphocyte proliferation and differentiation and finally antibody synthesis. Results underscore the

importance of sleep for immunocompetence.

S. Lanquillon1, I. Reckemeyer2, H. Vedder2, J.C. Krieg2, H. Klein1

1Department of Psychiatry, Regensburg University, Regensburg, Germany 2Department of Psychiatry,

Marburg University, Marburg, Germany

51. Cytokines - immunological link between Major Depressive Disorder (MDD) and alcohol abuse?

While some patients with MDD consume alcohol to gain relief of their complaints, in others depressive

symptoms even worsen by ethanol ingestion. Neither is it known by which pharmacological mechanisms

ethanol might influence depressive symptomatology nor whether certain psychopathological patterns of

the disease are associated with a distinct neurobiological reaction to ethanol. As ethanol strongly sup-

presses proinflammatory cytokine secretion in healthy people we measured the in vitro effect of ethanol

on cytokine production (etCP) in immune cells of blood samples from 37 patients with MDD and from 30

controls and correlated the results with clinical outcome and psychopathometry. We wanted to know

whether a) patients differ from controls with regard to ethCP b) the modulation of ethCP changes from the

pre-treatment status until remission c) ethanol effects correlate with specific psychopathological features.

There were no differences between patients and controls for TNF whereas ethanol suppressed IL-6 se-

cretion in untreated patients significantly less (p<0.01). After antidepressive treatment, suppressibility of

all patients' IL-6 secretion had returned to controls' values. While global depressive psychopathology was

weakly correlated with cytokine values, high scores on anxiety-specific scales were associated with in-

creased ethanol-induced suppression of IL-6 secretion (r=0.58, p<0.05). The correlation of increased

ethanol-induced suppressibility of IL-6 secretion with the degree of anxiety in MDD may be an immuno-

pharmacological link to relief-seeking drinking in MDD.

V.I. Leussink, B. Badent1, S. Veit1, B. Olshausen2, G. Becker3, M. Flentje2, B. Allolio1, W Arlt1

1Departments of Neurology, Endocrinology, and 2Radiotherapy, University of Würzburg, 3Department of

Neurology, University of Homburg, Germany

52. Time-dependant endocrine dysfunction and impact on well-being after radia-tion therapy of primary brain tumours and the influence of hormone substi-tution - a longitudinal study

Radiation-induced endocrine dysfunction and impact on well-being after treatment of primary brain tu-

mours is frequent and mostly due to hypothalamic damage. The aim of this longitudinal study was to in-

vestigate the hypothalamic-pituitary function of 31 long-term survivors (21m, 10f, age 29-65yrs) of primary

brain tumours (WHO II-IV) after surgery and radiation therapy (mean total dose 60.5 ± 5.2 Gy) and to

evaluate the influence of hormone substitution on well-being and quality of life. Endocrinological assess-

ment included measurements of thyroid, adrenal, and gonadal hormones at baseline and stimulating

tests. Validated questionnaires (SCL-90-R, GBBI-24, B-L von Zerssen, EORTC) were used for evaluation

of well-being. In patients with endocrine dysfunction (ED, n=14), endocrinological testing at time point 0

revealed significant lower levels of fT4 (14 ± 4 vs. 18 ± 6pmol/l in patients without ED (n=17), p<0.05) and

of DHEAS (684 ± 564 vs. 1439 ± 815ng/ml, p<0.01). After one year, fT4 serum levels of those patients

with thyroid hormone replacement therapy (n=5) remained stable, whereas there was a significant de-

crease of fT4 in those patients without treatment (n=14, 18 ± 4 vs. 14 ± 3 pmol/l). Measurement of serum

DHEAS levels in 21 patients (15m, 6f) revealed a significant decrease from 1130 ± 779 to 893 ± 568ng/ml

after one year. Psychometric tests revealed a striking increase of impairment of well-being in those pa-

tients without hormone replacement therapy (total complaint scores: GBBI-24 12.2 ± 12.2 vs.16.9 ± 14.4,

p<0.01, n=13). Due to a high prevalence of hypothalamic malfunction associated with endocrine dysfunc-

tion and impairment of well-being, yearly evaluation of the endocrine function and, if necessary, early

hormone replacement therapy should be recommended in patients being treated for malignant brain tu-

mours.

M. Levite

The Sackler School of Medicine, Tel Aviv University, and The Weizmann Institute of Science, Rehovot,

Israel

53. Nervous Immunity: Neurotransmitters, through their specific receptors, dic-tate T-cell functions, in a direct, powerful and contextual manner

Can activation of T-cell functions occur by means other than the traditional immunological signals? I ar-

gue that the nature and complexity of what T-cells have to deal with, and the speed at which decisions

have to be made, can not be fully satisfied by classical immunological signals such as antigens, cytokines

and chemokines. What other body means can assist in rapidly activating, regulating and orchestrating T-

cell activities in physiological and pathological conditions? I propose that powerful and direct signals are

supplied to T-cells by the highly branched nervous system, via specific neurotransmitters. Studying the

proposed direct functional dialogues between neurotransmitters and T-cells, my collaborators and I made

a series of observations, showing that several neurotransmitters (among them dopamine, Substance P,

CGRP, Neuropeptide Y, Somatostatin and GnRH), by direct binding to their cognate receptors on the T-

cell surface, dictate various T-cell functions in a very powerful manner. The T-cell features found to be

either activated or suppressed by neurotransmitters are as follows: gating of potassium channels,

changes in the T-cell membrane potential, integrin- mediated functions, cytokine secretion, in vivo behav-

iour, and de novo synthesis of genes, some of which not known to be expressed in T-cells. Taken to-

gether, I suggest that the nervous system, through specific neurotransmitters, can directly affect numer-

ous T-cell mediated immune functions and may enable direct neural-immune dialogs in health, stress,

injury and disease.

S.M. McCann, W.H. Yu, S. Karanth, C.A. Mastronardi

Pennington Biomedical Research Center (LSU), Baton Rouge, Lousiana USA

54. Control of the release of leptin, tumor necrosis factor alpha and nitric oxide by the central nervous system

Evidence is accumulating that the release of cytokines and nitric oxide (NO) is primarily controlled by the

central nervous system (CNS), although there is also local control exercised in the tissues that produce

these powerful mediators. There is a circadian rhythm of leptin release in man and rat with peak values at

1:30 a.m. which appears to be partly controlled by secretion of prolactin since prolactin stimulates leptin

release and the inhibitor of prolactin secretion bromocriptine lowers it. The levels of nitrate/nitrite

(NO3/NO2) in plasma reflects the production of NO throughout the body and the circadian rhythm of

NO3/NO2 parallels that of leptin suggesting that leptin may be mediating the rhythm of NO production.

Indeed, incubation of leptin with epididymal fat pads induces the release of NO. Anaesthesia causes a

decline in both plasma leptin and NO3/NO2 providing further evidence of neural control of both sub-

stances. Bacterial lipopolysaccharide (LPS), which causes responses in the body that mimic those of

infection causes a gradual release of leptin, that can be inhibited by bromocriptine or dexamethasone,

and by inhibition of alpha-adrenergic and beta-adrenergic inhibitory control over leptin release. Tumor

necrosis factor (TNF)-alpha responds rapidly to LPS, and in contrast to leptin, although this response is

blocked by anaesthesia and a beta-adrenergic agonist, there appears to be a stimulatory, instead of in-

hibitory alpha-adrenergic control involved in the response. Not only is TNF release stimulated by LPS, but

it is remarkably stress-responsive showing a 400-fold increase to the minor procedure of external jugular

catherisation. This response is delayed by the anaesthesia during the surgery. Stress induces a dramatic,

rapid decline in plasma (NO3/NO2), not related to the slight decline in plasma leptin that may be caused

by a neuronally mediated inhibition of NOS. That the central control is supplemented by local control is

shown by the fact that incubation of epididymal fat pads with leptin not only increases NO production, but

at lower concentrations activates TNF release. (This work was supported by NIH grant MH51853.)

H.-M. Meinck1, P. Henningsen2, C. Körner1, K. Dinkel3, W. Richter3

Departments of 1Neurology, 2Psychosomatic Medicine and 3Orthopedic Surgery, Ruprecht-Karls-

University, Heidelberg, Germany

55. Stiff man-syndrome, a neuropsychiatric disorder with autoimmunity against GABAergic neurones. Part I: Clinical, psychological and immunological characteristics

Stiff man-syndrome (SMS) and its variants have in common both, stiffness and spasms of the trunk mus-

cles and a dominant autoimmunity against GABAergic neurones. In a total of 68 patients, 33 were diag-

nosed with SMS, six with the minus-variant, stiff leg-syndrome (SLS), and 29 with the plus-variant, pro-

gressive encephalomyelitis with rigidity and myoclonus (PERM), respectively. Prevalence of serum auto-

autoantibodies against glutamic acid decarboxylase (GAD-Ab) ranged between 70 and 83 %. The core

symptoms, stiffness and spasms were often associated with gait disturbance, skeletal deformities and

increased startle reactions (70-100%). Acute autonomic disturbance (60-70%) could require intensive

care (20%) and cause sudden death (10%). Neurological reflex abnormalities so far not described were a

loss of the abdominal skin reflexes (40%) and a brisk head retraction reflex (45%). About 50% of patients

suffered from other organ-specific autoimmune diseases (most often type 1 diabetes mellitus) that had

manifested up to 20 years before or after the onset of neurological symptoms. Psychiatric evaluation re-

vealed moderate or severe life events preceding the onset of the disease by less than two years in 67%,

and development of task-specific agoraphobia in 64%. The cerebrospinal fluid (CSF) was abnormal in

64% comprising oligoclonal bands in 50%. Calculation of the antibody specificity index suggested inten-

sive intrathecal de novo synthesis of GAD-Ab. We conclude that SMS, SLS and PERM are manifesta-

tions of an immune-mediated chronic encephalomyelitis with characteristic motor, vegetative, and psy-

chological symptoms and signs. There is some relationship with other organ-specific autoimmune dis-

eases that are associated with GAD-Ab. However, intrathecal de novo synthesis of GAD-Ab suggests that

GAD-Ab may play a more specific pathogenetic role.

L.E. Miller1, J. Grifka, J. Schölmerich, R.H. Straub

Department of Internal Medicine and 1Orthopedic Surgery, University Hospital Regensburg, Germany

56. Norepinephrine from synovial tyrosine hydroxylase positive cells is a strong indicator of synovial inflammation in rheumatoid arthritis

Recently, we have demonstrated that density of sympathetic nerve fibres in synovial tissue was signifi-

cantly lower in patients with rheumatoid arthritis (RA) as compared to osteoarthritis (OA). This was ac-

companied by marked norepinephrine (NE) release from synovial tyrosine hydroxylase positive cells (TH+

cells). Synovial tissue of 34 patients with RA and 36 patients with OA was characterised using immuno-

histochemistry and a synovial tissue superfusion technique, respectively. In separate culture experiments

with mixed synoviocytes, the effect of NE on secretion of interleukin (IL)-6, IL-8, tumor necrosis factor

(TNF), and matrix metalloproteinase-3 (MMP-3) was investigated. Tissue density of TH+ cells was higher

in RA as compared to OA (63.9 vs 34.2 cells/mm(2), p=0.017) which was also found for synovial cellular-

ity, T cells, CD163+ macrophages, and cells of the synovial lining layer. Basal NE release from synovial

tissue highly significantly correlated with density of TH+ cells in RA (R=0.573, p= 0.001) but not in OA.

Basal NE release correlated with the degree of inflammation in RA (R= 0.420, p= 0.021) but not in OA,

and with spontaneous IL-8 secretion in RA (R= 0.581, p= 0.001) but not in OA. Only in RA, density of

TH+ cells positively correlated with spontaneous secretion of IL-6, IL-8, and MMP-3. We confirmed the

extensive loss of sympathetic nerve fibres in RA as compared to OA (0.32 vs 3.1 nerve fibre / mm(2),

p<0.001). TH+ cells and release of NE are strongly linked to a higher degree of synovial inflammation.

Culture experiments indicate that NE has antiinflammatory properties at higher concentrations (10(-6)M).

NE secretion of TH+ cells will probably be an antiinflammatory mechanism to counteract local inflamma-

tion. Thus, TH+ cell - derived NE is an important local factor of immunomodulation in synovial inflamma-

tion. Supported by the DFG (Str 511/5-1,2,3).

C. Nassenstein1, J.C. Virchow Jr1, S. Mayer1, H. Renz2, W. Luttmann1, A. Braun3

1Department of Pneumology, Medical University Clinics, Freiburg, Germany, 2Department of Clinical

Chemistry, University Clinics, Marburg, Germany, 3Fraunhofer Institute for Toxicology and Aerosol Re-

search, Hannover, Germany

57. Neurotrophin-4: Evidence for a proinflammatory role in allergic asthma?

Bronchial asthma has been defined as a chronic inflammatory disorder characterised by an increased

responsiveness of the airways to many non-specific stimuli. The neurotrophins NGF, BDNF, NT-3 and

NT-4 are enhanced in atopic diseases and seem to initiate bronchial hyperresponsiveness (BHR). How-

ever, limited information about the cellular sources of NT-4 and its proinflammatory potency in asthma are

available. Therefore, we investigated local synthesis of NT-4 in the lung of asthmatics. Thus, broncho-

scopy and bronchoalveolar lavage (BAL) following segmental allergen provocation (SAP) were per-

formed. RT-PCR and immunohistochemical staining of BAL-cells indicate an NT-4 synthesis by eosino-

phils and macrophages. To examine a possible role in inflammation, eosinophil function in response to

NT-4 was studied in vitro. Viability was assessed with propidium-iodide whereas activity was measured

by flow-cytometry using CD69- antibodies. Stimulation of purified blood-eosinophils with NT-4 does not

influence survival and activation while incubation with NT-4 of eosinophils obtained from BAL fluid (BALF)

after SAP prolongs viability and increases CD69-expression. These data indicate that the release of NT-4

in asthma following allergen challenge might link the inflammatory changes observed in asthma with the

increase in BHR. Therefore, antagonisation of the NT-4 signal could be a promising concept for future

therapeutic strategies in allergic asthma.

H. Neumann

Neuroimmunology, Max-Planck-Institute of Neurobiology, Martinsried and Neuroimmunology, European

Neuroscience Institute Göttingen

58. Selective attack of MHC class I-induced neuritis by cytotoxic T lymphocytes Cytotoxic T lymphocytes (CTLs) invading the brain tissue could damage neurons during neuroinflamma-

tory diseases. In multiple sclerosis transection of axons and spheroid formation is associated with the

demyelinating and inflammatory process. Cultured murine hippocampal neurons were treated with IFN-g

and tetrodotoxin to induce MHC class I and Fas molecules. The neurons were confronted with CTLs and

the mode of cell death induced in neurons was analysed by continuous measurements of intracellular

calcium (Ca2+)i levels and visualisation of the cell membrane. Neurons attacked by CTLs did not show

the typical perforin-mediated, early (Ca2+)i responses that were readily detected in astrocytes. In con-

trast, neurons showed a gradual and sustained increase in (Ca2+)i within 2-4 hours following CTL attack

that was mediated by Fas molecules. While the neuronal soma was protected against early perforin-

mediated CTL cytotoxicity, the neurites rapidly developed cytoskeleton breaks with adjacent spheroids.

The CTLs induced segmental membrane disruption 5-30 minutes after the establishment of CTL-neurite

contact, as demonstrated by continuous confocal analysis of fluorescence-labelled plasma membranes.

Transection of neurites was only detected when the neurons were pulsed with the peptide recognised by

the CTLs. Our results show that the neuronal soma is protected against early perforin-mediated CTL at-

tack. In contrast, MHC class I/ peptide-restricted CD8+ CTLs can induce lesions to neurites that might be

responsible for axonal damage in autoimmune and neurodegenerative CNS diseases.

I.D. Neumann1, S. Krömer2, A. Wigger2

1Institute of Zoology, University of Regensburg, Regensburg, Germany; 2Max Planck Institute of Psychia-

try, Munich, Germany

59. Early life stress affects the emotional and neuroendocrine development of rat offspring in dependence on the innate level of anxiety

Pre- and postnatal exposure to stressful events results in chronic alterations of emotionality and neuroen-

docrine stress responsiveness. Using two rat lines selectively bred for high (HAB) and low (LAB) anxiety-

related behaviour, respectively, we test the hypothesis whether behavioural and neuroendocrine conse-

quences of pre- and postnatal stress depend on the genetically determined level of anxiety. For prenatal

stress, between days 6 and 20 of pregnancy, HAB and LAB rats were daily exposed to a psychosocial

(maternal defeat) and emotional (restraint) stressors. For postnatal stress, pups were daily separated

from their mother for 3 hours. Interestingly, differences in the emotionality between HAB and LAB rats

persist in pregnancy and lactation. At the age of 12 weeks, the emotionality of the offspring was tested,

for example, on the elevated plus-maze and the modified holeboard. Our preliminary results indicate that

pre- or postnatally stressed HAB rats are characterised by a reduced, rather normal, emotionality. Fur-

ther, the reactivity of the hypothalamo-pituitary-adrenal axis was tested under basal conditions and in

response to a mild, emotional stressor. In HAB rats, a pathologically high stress reactivity of the HPA axis

was described before and confirmed in unstressed controls of our study. In postnatally stressed HAB rats,

HPA axis responses were found to be rather reduced. Thus, consequences of early life experiences on

emotional and neuroendocrine stress response in adulthood depend on the “inborn“ level of anxiety indi-

cating differences in the vulnerability to negative environmental factors. Supported by DFG.

J.H. Niess1,3, A. Dignass2, A.C. Zenclussen1,3, R. Joachim1,3, H. Mönnikes2,3, B.F. Klapp3, P. Arck1,3

1Biomedical Research Center, Campus Virchow-Clinic, Berlin, Germany 2Department of Hepatology and

Gastroenterology, Campus Virchow-Clinic, Berlin, Germany 3Department of Psychosomatics, Charité,

Berlin, Germany

60. Influence of stress on experimentally induced murine colitis In their daily work many physicians are implicating that stress influences inflammatory bowel diseases

(IBD). Recently, various studies revealed that the risk of relapses is related to stress, i.e. the exacerbation

of experimentally induced colitis could be triggered by exposing mice to stress. Further, in humans long-

term perceived stress was shown to be related to relapses of ulcerative colitis. Therefore the aim of our

study was to identify immune mediators involved in stress-induced effects on the mucosa. In Balb/c mice

colitis was induced by intrarectal application of 36 micro-gram DNBS and by exposing Balb/c mice to a

ultrasonic stress source. On day 5 or 9 after induction of colitis distal colon was prepared. CD4+, CD8+

and gamma delta TCR+ cells were detected by immunohistochemistry. The epithelial integrity was char-

acterised by detecting apoptotic signals (TUNEL) and by staining for the proliferation marker Ki67. On day

5 and day 9 after induction of colitis an increase of CD4+, CD8+ and TUNEL+ and a reduction of Ki67+

signals were detected in samples obtained from colitis exposed to stress compared to non-stressed mice.

An increase of gamma delta TCR+ cells was only observed on day 5 after induction of colitis compared to

non-stressed mice. In further studies it would be of interest to identify the mediators involved in the stress-

triggered brain-gut axis. Tachykinins have been suggested to mediate stress induces alterations of IBD,

and therefore the stress induced attack on the mucosal integrity might be abrogated by the treatment of

IBD with neurokinin antagonists.

R. Oberbeck1, D. Schmitz1, M. Schüler, K. Wilsenack, C. Biskup, M. Schedlowski, D. Nast-Kolb1, & M.S.

Exton

Department of Medical Psychology and 1Emergency Surgery, University Clinic of Essen, Germany

61. Catecholaminergic modulation of cellular immune function and survival dur-ing chronic sepsis in the mouse

A pronounced increase of catecholamine concentration in blood is observed during sepsis. Immunomodu-

lating effects of catecholamines are reported, particularly for adrenaline and noradrenaline. Those effects

are primarily mediated by beta-adrenergic receptors. The aim of our study was to therefore investigate

the impact of adrenergic substances on survival and immune function of septic mice, 48h after sepsis

induction. Sepsis was induced by ceacal ligation and puncture (CLP) in male MNRI mice. Immune status

was assessed by measuring the rate of apoptosis (FACScan) and the proliferative capacity of spleno-

cytes (H3-Thymidin intake), as well as cytokine production (ELISA). Also, leukocyte distribution in periph-

eral blood was monitored by FACScan. The animals were treated in four groups. The first group received

an continuous application of 0.06mg/kg/h noradrenaline via an intraperitoneal osmotic pump. The second

group received 0.5mg/kg propranolol by intraperitoneal injection each 12h. The third group received a

combination of noradrenaline and propranolol while the fourth group received NaCl i.p. CLP-induced sep-

sis produced 50% lethality. Noradrenaline decreased lethality of septic mice. However, propranolol alone,

or in combination with noradrenaline increased lethality. In contrast, all treatments produced an elevation

of splenocyte apoptosis, as noradrenaline increased apoptosis, propranolol raised it further and the com-

bination of both substances produced the highest apoptosis rate. Sepsis produced a dramatic leukopenia

in peripheral blood that was observed for all cell subpopulations. The proliferative capacity of splenocytes

was decreased by CLP induced sepsis, and further decreased by noradrenaline. An even stronger de-

pression was revealed by propranolol treatment, however the combination produced no further increase.

The effect of sepsis and adrenergic treatment of splenocyte proliferation was mirrored by pronounced

decreases in IL-2 and IFN-gamma production. Thus, an adrenergic-receptor mediated influence on the

cellular immune function during a systemic inflammatory reaction is possible. This should be regarded

during the treatment of septic shock.

G. Pacheco-López, M. Schedlowski

Department of Medical Psychology, University of Essen, Germany

62. Central noradrenergic depletion reduces brain c-Fos expression to a protein antigen

Peripheral administration of antigens has been shown to activate induction of Fos in the brain, but the

mechanism is not known. Since cerebral noradrenergic system has been implicated in Fos induction, we

studied the protein antigen-induced appearance of Fos in rats pre-treated with 6-hydroxydopamine (6-

OHDA), which depleted cerebral norepinephrine (NE), or vehicle. All the animals received i.p. injections of

Keyhole Limpet Hemocianin (KLH) in a dose that did not produce side effects but was sufficiently immu-

nogenic. Previously, we have demonstrated that this immune stimulus increased c-Fos in several brain

regions, i.e., in the hypothalamic paraventricular nucleus (PVN) and the central nucleus of amygdala

(CeA), parabrachial nucleus (PBN) and nucleus of the solitary tract (NTS). Pre-treatment with 6-OHDA in

the brain lateral ventricles, substantially reduced the KLH-induced Fos in all those brain areas. These

results suggest that the noradrenergic innervation of the forebrain (PVN and CeA) coming from the brain-

stem (NTS and PBN) are involved in this immune to brain communication phenomenon.

G. Päth, A. Braun1, N. Meents, S. Kerzel, U. Raap, G.W. Hoyle2, WA Nockher and H. Renz

University Hospital, Department of Clinical Chemistry and Molecular Diagnostic, Marburg, Germany;

1Fraunhofer-Institute of Toxicology and Aerosol Research, Hannover, Germany; 2Section of Pulmonary

Disease, Critical Care and Environmental Medicine, Department of Medicine, Tulane University Medical

Center, New Orleans, USA

63. Allergic early-phase reaction and sensory reactivity are enhanced in trans-genic mice with lung-specific overexpression of NGF

Nerve growth factor (NGF) acts as an amplifier of allergic airway inflammation and plays a role in the

development of airway hyperreactivity. Furthermore, NGF is reported to induce degranulation of mast

cells in vitro. Allergen-induced mast cell mediator release is the main event causing allergic early-phase

reaction. Therefore, we tested the in vivo-relevance of this finding in a murine model of asthma utilising

mice with constitutive lung-specific overexpression of NGF under the control of a Clara-cell secretory

promoter (CCSP-NGF-tg) and wildtype (wt) controls. Prior to experiments, mice were sensitised to and

challenged with ovalbumine (OVA). Compared to wt controls, allergic CCSP-NGF-tg mice display an in-

creased allergen-induced airflow limitation. NGF-overexpression did not alter the serum levels of IgE an-

tibodies in allergic mice but increased the release of serotonin, which is the main mediator of the allergic

early-phase reaction in rodents. Moreover, NGF-overexpression increased the reactivity of sensory neu-

rons in response to inhaled capsaicin. In conclusion, NGF enhances both allergen-dependent early-phase

reaction and allergen-independent airway hyperreactivity and therefore, appears as an important media-

tor of neuro-immune interactions in the lung during allergic airway inflammation and asthmatic responses.

Supported by the Volkswagen-Foundation

M. Peters1,2, M. Rothermundt1, N.T. Bui2, V. Arolt1, J.H.M. Prehn2

1Department of Psychiatry and 2Interdisciplinary Center of Clinical Research of the Medical Faculty, Jun-

ior Research Group, University of Münster, Germany

64. S100B protects against neuronal cell death induced by NMDA or hypoxia

The acidic Ca2+ binding protein S100B is produced mainly by astrocytes. It can exert neuroprotective

action on neurons but is also able to induce apoptotic cell death. In clinical studies S100B levels are ele-

vated in patients with schizophrenia and melancholic depression. It remains to be clarified whether the

increase of S100B in the psychiatric conditions represents a neurodegenerative or neuroregenerative

response. In order to improve the knowledge about the physiological functions of S100B in the CNS, we

investigated the effects of S100B in cultured primary neurons of rat hippocampus. Primary hippocampal

neurons were derived from neonatal rats and cultured for 12-14 days under serum-reduced conditions.

Immediately prior to an exposure to NMDA (0.3 mM, 5 min) or hypoxia (95 % N2, 5 % CO2 for 24 h) they

were treated with S100B (2-200 ng/ml). After twenty-four hours the percentage of damage was assessed

by the trypan blue dye-exclusion method. Treatment with 200 ng/ml S100B significantly reduced the toxic

effects of both NMDA (NMDA-exposed controls: 33 ± 9.3 % cell death; NMDA plus S100B: 8.7 ± 3.9 %;

n=8; p<0.01) and hypoxia (Hypoxia-exposed controls:25.9 ± 13.9 % cell death; Hypoxia plus S100B: 10.8

± 5.8; n=12; p<0.01). There was no significant difference in the amount of cell death compared to control

condition (sham-exposure and normoxia, respectively). The neuroprotective effect of S100B was not

dose-dependent at the chosen range of concentrations. In primary hippocampal neurons S100B has po-

tent neuroprotective properties against excitotoxic and hypoxic cell death.

U. Renner, P. Lohrer, M. Tichomirova, M. Paez Pereda, L. Schaaf, E. Arzt, G.K. Stalla

Department of Endocrinology, Max-Planck-Institute of Psychiatry, Munich, Germany

65. Expression and role of Toll-receptors in pituitary folliculostellate cells Folliculostellate (FS) cells represent about 5 to 10 % of all pituitary cells. They do not produce hormones

but IL-6, which is stimulated by TNF or interleukin-1 (IL-1). Therefore FS cells are considered to be in-

volved in immune-endocrine cross-talk during bacterial infection. We have studied whether FS cells ex-

press functional active Toll-receptors type 4 (Tlr4) and 9 (Tlr9) which mediate cytokine induction in re-

sponse to LPS (Tlr4) or bacterial DNA (Tlr9). In the folliculostellate TtT/GF mouse pituitary tumour cell

line and in mouse ptuitary both Tlr4 and Tlr9 expression could be demonstrated. Whereas the effect of

bacterial DNA on IL-6 production and its mechanism is still under investigation, we could already demon-

strate that LPS directly stimulates IL-6 secretion by TtT/GF cells and normal FS pituitary cells. FS cells do

not only express Tlr4 but also CD14 which is known to improve the binding and subsequent action of

LPS. LPS-induced IL-6 production in FS cells involves stimulation of phosphoinositol turnover, phos-

phorylation of p38 alpha MAP kinase, phosphorylation of inhibitor of kappa-B and subsequent transcrip-

tional activation of nuclear factor kappa-B. In three-dimensional histotypic pituitary re-aggregates, LPS

was able to stimulate ACTH secretion via paracrine-acting FS cell-derived IL-6. In this way, LPS-

stimulated intrapituitary IL-6 might contribute to the activation of the HPA axis during bacterial infection.

H. Rogausch, A. Del Rey, D. Zwingmann, K. Voigt, H. Besedovsky

Institute of Physiology, Philipps-University, Marburg, Germany

66. The regulation of blood pressure is affected by an IL-1-mediated re-setting of baroreceptor reflexes at CNS levels

Cardiovascular adjustments occur during diseases involving the activation of the immune system, and it is

known that IL-1 beta can affect cardiovascular functions by influencing the activity of the autonomic nerv-

ous system at central and peripheral levels. We hypothesised that IL-1 plays an important role in such

adjustments by re-setting reflex mechanisms that stabilise arterial blood pressure at central levels. This

hypothesis was tested by studying whether the baroreceptor reflex is affected by endogenous cytokines

produced following intravenous injection of the endotoxin LPS. The reflex was evoked by administration

of phenylephrine. The sensitivity of the reflex was calculated from the slope between cardiac inhibition

and increased arterial blood pressure. The efficacy of the reflex in controlling peripheral vascular resis-

tance was evaluated by integrating changes in peripheral resistance following phenylephrine administra-

tion. The results indicate that the gain of the reflex increased significantly 2h after LPS administration, i.e.

the sensitivity to counterbalance changes in arterial blood pressure was increased, but that its activation

was significantly shortened. These changes tended to stabilise blood pressure at a higher level, as indi-

cated by the increase in peripheral vascular resistance observed when the reduction of plasma volume

induced by LPS was compensated. Alterations of the sensitivity and efficacy of the reflex were eliminated

by chronic i.c.v. infusion of IL-1ra (500 microg/day, 5 days). It is concluded that peripheral administration

of LPS induces complex changes in cardiovascular reflexes, and that IL-1 beta contributes to control

blood pressure by re-setting the sensitivity of baroreceptor reflexes at brain levels. Supported by the

Volkswagen-Foundation and the SFB 297.

N. Rohleder, J.M. Wolf, C. Kirschbaum

Institute of Experimental Psychology II, University of Düsseldorf, Germany

67. Impact of gender and oral contraceptive use on glucocorticoid sensitivity of pro-inflammatory cytokine production after psychosocial stress

Women and men show large differences in susceptibilities to immune disorders. These gender differ-

ences may be mediated by functional interactions of the hypothalamus-pituitary-adrenal (HPA) and the

hypothalamus-pituitary-gonadal (HPG) axes. A potential mechanism involved in this interaction is the

glucocorticoid (GC) sensitivity of relevant target tissues. We investigated HPA axis activation and GC

sensitivity of pro-inflammatory cytokine production after psychosocial stress in healthy young women and

men. A total of 58 subjects were investigated in the present study. Eighteen women in the luteal phase of

the menstrual cycle, 13 women using oral contraceptives (OC), and 27 men were exposed to a psycho-

social stress test (TSST). Free cortisol was measured repeatedly before and after stress. GC sensitivity

was assessed by dexamethasone (DEX) inhibition of lipopolysaccharide (LPS) stimulated production of

interleukin-6 (IL-6). The stress test induced significant increases in free cortisol with no differences be-

tween men and luteal phase women, while OC users showed blunted cortisol responses. GC sensitivity

showed large gender differences: In men GC sensitivity markedly increased one hour after stress, while

GC sensitivity in luteal phase women slightly decreased. Women using OC showed response patterns

comparable to men. These results demonstrate, that despite similar free cortisol responses of men and

luteal phase women to stress, gender may exert differential effects on the immune system by modulating

the GC sensitivity of pro-inflammatory cytokine production. OC use seems to compensate the lower corti-

sol levels after stress by increasing GC sensitivity of pro-inflammatory cytokine production.

M. Rothermundt1, M Peters1, M. Wiesmann2, S Rudolf3, M Hettich1, S. Abel1, H. Kirchner3, V. Arolt1

1Department of Psychiatry, University of Münster School of Medicine, Münster, Germany, 2Department of

Neuroradiology, Medical University of Lübeck, Germany, 3Institute of Immunology and Transfusion Medi-

cine, Medical University of Lübeck, Germany

68. S100B in schizophrenic psychosis

S100B, a calcium binding cytokine, is produced mainly by astroglial cells. It is involved in a variety of cel-

lular mechanisms such as the regulation of energy metabolism, cellular proliferation and differentiation. It

stimulates microglial cells, impairs the proliferation of cytotoxic T-cells and decreases the secretion of

interferon gamma. Secreted in nanomolar concentrations it promotes neuronal survival and regeneration.

Micromolar concentrations, however, evolve neurotoxic activity and induce the apoptosis of neurons and

glial cells. In adult brains it plays a role in neuronal plasticity and long term potentiation. S100B has been

shown to be increased in traumatic brain injury, toxic or ischaemic brain damage, in multiple sclerosis and

neurodegenerative disorders. A recent study showed increased S100B levels in medicated acutely psy-

chotic patients with schizophrenia. The study presented here included 26 drug-free patients with acute

schizophrenia and 26 matched healthy controls. S100B blood concentrations were determined using a

quantitative immunoassay upon admission and after 6 weeks of neuroleptic treatment. The PANSS was

used to investigate psychopathology. Unmedicated schizophrenic patients showed significantly increased

S100B levels compared to matched healthy controls. After 6 weeks of treatment, 11 patients showed

normal S100B levels while in 15 patients the levels remained increased. These patients showed signifi-

cantly higher PANSS negative scores upon admission and after 6 weeks of treatment. Schizophrenic

patients display ongoing neuroplastic activity in acute psychosis. Continuously increased S100B levels

are associated with the persistence of negative symptomatology.

K. Ruprecht, V. Hummel, P. Rieckmann

Department of Neurology, Julius-Maximilians-University, Würzburg, Germany

69. Production of Oncostatin M by human microglia and peripheral blood mono-nuclear cells: Potential sources of a cytokine with immunomodulatory ef-fects on human cerebral endothelial cells

We have recently shown that oncostatin M (OSM), a member of the IL-6 family of cytokines, has potent

immunomodulatory effects on human cerebral endothelial cells in vitro. Furthermore, expression of OSM

was detected immunohistochemically in association with inflammatory infiltrating cells, reactive astro-

cytes, and microglia in brain lesions from patients with multiple sclerosis. This suggested that OSM may

participate in the pathophysiology of inflammatory brain disease, possibly via immunoregulatory functions

in human cerebral endothelial cells. Here, we aimed to further characterise potential extra- and intracere-

bral sources of OSM known to interact with cerebral endothelial cells of the blood-brain barrier (BBB). We

therefore studied in vitro production of OSM by cultured human microglia and peripheral blood mononu-

clear cells (PBMC) under basal conditions and after treatment with various stimuli using ELISA tech-

niques. Human microglia secreted low baseline levels of OSM, however, stimulation with LPS induced

OSM production in microglial cells. IFN-gamma and amyloid-beta [1-42] failed to upregulate OSM in mi-

croglial cells. PBMC were found to release certain levels of OSM spontaneously. In contrast, human

cerebral endothelial cells did not produce OSM neither under basal conditions nor after treatment with

proinflammatory stimuli (LPS, IFN-gamma, TNF). These results suggest that activated microglia may be

an endogenous source of OSM within the CNS. OSM may thus represent a signal by which microglial

cells interact with the endothelium of the BBB. At the same time, OSM produced by PBMC may influence

BBB function from the extracerebral side.

M.J. Schwarz, M. Riedel, H. Krönig, S. Chiang, M. Ackenheil, N. Müller

Psychiatric Hospital, University of Munich, Germany

70. Th1 and Th2 relationship in schizophrenia - immunological, immunogenetic and therapeutic aspects

Immunological abnormalities in a subgroup of schizophrenia have frequently been reported. After review-

ing the literature, we have hypothesised a decrease of the Th1-like (cell-mediated) immune system and

increased activation of the Th2-like (humoral) immune system in a distinct group of schizophrenic pa-

tients. Based on this Th2-hypothesis of schizophrenia, we have performed several biochemical and ge-

netic investigations and have carried out a therapy study, using a drug, which is known to induce a switch

from Th2 to Th1 predominance. Investigation of cytokine production by in-vitro stimulated lymphocytes

using ELISPOT technology. Molecular genetic investigations (schizophrenic patients n= 150; healthy

controls n=170) of candidate genes coding for immunologically relevant genes - especially those, in-

volved in Th1/Th2 balance (IL-12, IL-13, IFN-gamma, IL-4 etc.) or those, located on hot spots, identified

by genome scans (chromosome 6 - TNF, chr. 22 - IL-2R, chr. 4 - IL-2). Clinical study in a placebo-

controlled, cross-over, add-on design, including 50 schizophrenic patients using Celecoxib added to an

antipsychotic medication with Risperidone. ELISPOT: preliminary results suggest a subgroup of schizo-

phrenic patients with reduced IFN-gamma production and increased IL-4/IL-13 production. Immunogenet-

ics: The functional A1082G promotor polymorphism was more frequent in patients compared to controls.

Therapy study: Patients who received Celecoxib showed a markedly faster reduction of psychotic symp-

toms, than those, who received the placebo together with Risperidone. Our complex but systematic re-

sults may have great impact for the identification of a subgroup of schizophrenia with immune-related

pathophysiology and for the development of an immune-mediated therapy strategy in schizophrenia.

M. J. Schwarz, N. Müller, M. Riedel, C. Scheppach, S. Sokullu, M. Ulmschneider, R. Yolken1

Psychiatric and Psychotherapeutic Hospital, Ludwig-Maximilian University Munich, Munich, Germany;

1Stanley Division of Developmental Neurovirology, Johns Hopkins Univ., School of Medicine, Baltimore

MD, USA

71. Are high anti - cytomegalovirus titres in schizophrenia associated with bet-ter outcome to anti-inflammatory therapy?

Inflammation may play a role in the pathophysiology of schizophrenia. Anti-inflammatory therapy with the

COX-2 inhibitor celecoxib added to the neuroleptic risperidone has been shown to improve the outcome

of schizophrenic patients. In order to analyse the possible influence of cytomegalovirus infection, the anti-

CMV antibody-titres were measured in patients treated with celecoxib and in those without celecoxib

treatment (placebo). There was no difference regarding the CMV titres between the two groups. In the

placebo-group (n = 23) there was no statistical significant relationship between the improvement of the

schizophrenic symptomatology measured by the positive and negative symptom scale (PANSS). How-

ever, in the group treated with celecoxib (n = 23) we found a statistical significant relationship between

the improvement on the PANSS-positive scale, the PANSS-global scale and the PANSS-total scale. Pa-

tients with higher anti-CMV titres had a better improvement of their schizophrenic symptoms. This result

implicates that anti-inflammatory therapy with celecoxib may have better effects in those schizophrenic

patients, in whom an infection with cytomegalovirus may play a role. However, statistical limitations have

to be discussed. Supported by the Vada and Theodor Stanley Foundation.

S. Sorge, T. Pollmächer, D.M. Lancel

Max-Planck-Institute of Psychiatry, Munich, Germany

72. Effects of clozapine on sleep and brain concentration of TNF in rats In psychiatric patients, treatment with the atypical antipsychotic clozapine is accompanied by sleepiness

and increased circulating levels of cytokines and soluble cytokine receptors. During night sleep, total

sleep time, sleep efficiency and the absolute and relative amounts of nonREM sleep, particularly stage 2,

are increased (1,2). The most prominent immunological effects of clozapine are increases in the plasma

concentrations of TNF and soluble TNF-receptors p55 and p75. Because TNF is a pivotal mediator of

sleep changes during infections (3), we hypothesised that the influence of clozapine on sleep might also

involve TNF. In the present vehicle-controlled study we investigated the influence of subchronic clozapine

treatment on sleep-wake behaviour and on the brain levels of TNF in rats. Clozapine consistently reduced

wakefulness and increased nonREM sleep, which was associated with prolonged nonREM episodes.

Furthermore, slow wave activity within nonREM sleep was attenuated. Initially, after acute administration,

REM sleep was markedly suppressed with a rebound afterwards. Clozapine produced a transient reduc-

tion in brain temperature which was followed by an increase. High doses of clozapine also elevated the

concentration of TNF in the frontal cortex. These findings suggest a strong and rapid action of clozapine

on sleep-wake regulation which maybe caused, among other effects of the drug, by a stimulating effect

on TNF production within the brain.

B. Sperner-Unterweger1, S Eder, B Widner, D Fuchs

1Department of Biological Psychiatry and Institute of Medical Chemistry and Biochemistry, University of

Innsbruck, Austria

73. Immunology and antidepressants: Modulatory effect of selective serotonin reuptake inhibitors on the human myelomonocytic cell line THP1

The essential amino-acid tryptophan plays crucial role in mainly two biochemical pathways: biosynthesis

of 5-hydroxy-tryptamine (serotonin) and the formation of kynurenine depending on tryptophan pyrrolase

(PT) or indoleamine 2,3 dioxygenase (IDO) enzyme activity. IDO is induced by interferon-γ (IFN- γ) and

therefore it seems to represent a link between immunological and neurotransmitter functions. Deficits in

the central serotonergic system are related to the development of depression. Selective serotonin reup-

take inhibitors are well established as antidepressant agents but little is known about this influences on

the IDO activity as well as about possible immunomodulatory effects. The aim of this study was to evalu-

ate possible modulatory effects on the IDO activity and/or immune activation due to citalopram. For this

investigation monocytic THP-1 cell were used measuring neopterin, tryptophan and kynurenine release

as well as calculating the kynurenine/tryptophan ratio. THP-1 liquid cell cultures were incubated with in-

creasing doses of citalopram (0, 20, 100, 1000 mg/ml). Furthermore a comparison between cultures

stimulated with and without IFN-gamma (100 units/ml) was performed. The above mentioned parameters

were evaluated after 24, 48 and 72 hours of incubation. In cell cultures without IFN-gamma stimulation,

upon addition of citalopram no significant differences of neopterin, kynurenine and tryptophan were seen

during the observation period. In cell cultures with IFN-gamma stimulation, citalopram induced a signifi-

cant dose-dependent increase of neopterin (after 24, 48 and 72 hours; p < 0.01). The same development

was found for kynurenine (24, 48 hours; p < 0.01) whereas tryptophan presented a dose-dependent sig-

nificant decrease (24, 48, 72 hours; p < 0.01). Kynurenine/tryptophan ratio showed a significant dose-

dependent increase (24, 48, 72 hours; p < 0.01). These results primarily indicate immunomodulatory ef-

fect due to the SSRI compound citalopram.

V. Stefanski, L. Dawils, H. Engler, G. Knopf, S. Kurth, A. Peschel, S. Schulz


74. Effects of psychosocial stress on immune cells in laboratory rats: endocrine und immunological mechanisms

Social stress in laboratory rats is associated with substantial alterations in the immune system. In this

report findings on dominant and subdominant males in social confrontations and on males living in natu-

ralistic colonies are summarised. Subdominant males in social confrontations show reduced functional

capacities of T cells as well as a decrease in the number of circulating T cells. Migration studies using

radiolabelled T cells demonstrate that blood T cells do not accumulate in lymphoid tissue or other body

compartments (e.g., skin, lungs, liver) of subdominant animals. On the other hand, social stress enhances

apoptosis of immature T cells and may therefore contribute to lower T cell numbers in these animals.

Glucocorticoid action accounts for the decrease in blood T cell numbers; however, suppression of T cell

proliferation was found largely unaffected in adrenalectomised as well as RU486 treated subdominant

males. Also unspecific immune functions such as NK cytotoxicity are modulated in socially stressed rats:

the suppression of NK activity is of clinical significance as a several-fold increased tumor metastasis oc-

curs (MADB106 tumor model). In contrast to subdominant males, initially similar immunological changes

in dominant males disappear when confrontations last for longer periods (1 week). Finally, data from rat

colonies indicate that immunological changes also occur in naturalistic social environments and persist

over weeks. Supported by the Volkswagen foundation (I75144).

H. Steffen1, N. Menger4, H. Krastel1, W. Richter3 and H.-M. Meinck2

1Departments of Ophthalmology, 2Neurology and 3Orthopedic Surgery, Ruprecht-Karls-University, Hei-

delberg, 4Max Planck Institute for Brain Research, Frankfurt, Germany

75. Stiff man-syndrome, a neuropsychiatric disorder with autoimmunity against GABAergic neurones. Part III: Abnormalities in retinal function linked to autoantibodies against glutamic acid decarboxylase (GAD)

In stiff man syndrome (SMS) and its variants, the presence of autoimmunity against the rate-limiting en-

zyme of GABA synthesis, glutamic acid decarboxylase (GAD) suggests widespread impairment of

GABAergic inhibition. As the retina contains numerous GABAergic neurones, we investigated retinal func-

tions in 20 SMS patients, none of them with visual complaints. Investigations comprised biomicroscopy,

conventional electroretinogram (ERG), pattern and multifocal ERG (mfERG), colour vision, dark adapta-

tion, and contrast sensitivity. Presence of antiretinal antibodies was analysed by indirect immunohisto-

chemistry on cryostat sections of macaque retina with serum from patients and controls and with mono-

clonal anti-GAD65 antibodies. Antibodies against GAD (GAD-Ab) were present in 13 of 20 patients. Ten

of them had abnormal ERG responses, and in five these physiological abnormalities were rated as se-

vere. In contrast only 3 out of 7 patients tested negative for GAD-Ab had ERG-abnormalities which were,

moreover, rated as severe in none. Retinal immunofluorescence revealed strong reactivity of the inner

plexiform layer and to a lesser extent of the outer plexiform layer with serum from patients tested positive

for GAD-Ab. An almost identical staining pattern was noted with monoclonal GAD65-Ab. No staining was

observed with serum from patients tested negative for GAD-Ab and with serum from healthy controls.

These results suggest that GAD-Ab in patients with SMS and their antiretinal antibodies are identical.

Electrophysiological retinal dysfunction was associated with the presence of GAD-Ab (p<0,04) suggesting

an autoimmune attack. Widespread GABAergic dysfunction seems to be a feature of SMS, but remains

subclinical in the majority of cases.

M. Stephan1, T. Breivik2, R.H. Straub3, R. Pabst1, S. von Hörsten1

1Departments of Functional and Applied Anatomy, Medical School of Hannover, Germany,

2Periodontology, Faculty of Dentistry, University of Oslo, Norway, 3Internal Medicine, University of Re-

gensburg, Germany

76. Nature-nurture interactions determine adult disease susceptibility: Stress protection as an option for the treatment of periodontal disease

Epidemiological studies suggest that a high susceptibility for periodontal disease is commonly associated

with an increased reactivity for stress in humans. The present studies investigated whether acquired dif-

ferences in the stress response interact with a given genetic background and whether interventional

strategies, known to reduce stress responsiveness, are successful in preventing increased disease sus-

ceptibility. Repeated maternal deprivation and postnatal handling of rats were used to either increase or

decrease stress responsiveness in adulthood. These different postnatal experiences were applied to in-

bred F344 and LEW rat strains that genetically differ in their stress responsiveness. Interventional strate-

gies consisted in either prophylactic-like additional juvenile tactile stimulation or in pharmacological treat-

ment with antidepressants in adult rats that had experienced maternal deprivation in infancy. Differences

in the individual stress responsiveness were monitored by a behavioural tests, by measuring corticoster-

one, NPY and cytokine plasma levels. Periodontal disease was induced by silk-ligatures around molar

teeth in adult rats. Stress high responding F344 rats developed significantly more periodontal disease.

Only in the stress low responding LEW rat strain, handling-induced reduction of adult stress responsive-

ness was protective while maternal deprivation aggravated experimental periodontal disease. Interven-

tions by additional tactile stimulation in infancy as well as antidepressants in adulthood protected from the

deprivation-induced aggravation of the disease. Genetic and non-genetic factors, which persistently in-

crease the individual responsiveness to stress, aggravate periodontal disease. It is suggested that stress

protection is effective in the treatment of periodontal disease in a subgroup of genetically stress low re-

sponding individuals. Supported by the Volkswagen-Foundation I/75169.

J.R. Stevenson, J. Westermann1, S. Supanz, P.M. Liebmann, K. Schauenstein

Institute of Pathophysiology, University of Graz, Austria, 1Institute of Anatomy, Medical University of

Lübeck, Germany

77. Alpha-adrenergic treatment differentially affects expression of adhesion molecules and leads to enhanced lymphocyte apoptosis in lymphoid organs of the rat

We have previously shown in the rat that peripheral catecholamines affect immune functions via alpha-

adrenergic mechanisms. One catecholamine effect is to change cell numbers in the different immune

compartments. To discover the mechanisms, we measured leukocyte numbers, expression of adhesion

molecules and rate of apoptosis in cells of different lymphoid organs. Rats were treated for 12 hours with

implanted tablets that continuously released noradrenaline and propranolol. Adhesion molecules were

measured by flow cytometry, and cell numbers by Coulter Counter, haemocytometer, and flow cytometer.

Apoptosis was determined on tissue sections by the TUNEL technique. The treatment caused reduced

expression of L-selectin on cells of blood and lymph nodes and an increase in the spleen. Furthermore, in

treated rats, the LFA-1 expression of granulocytes was decreased in all organs studied. Although granu-

locyte numbers increased in the blood, lymphocyte numbers were reduced in all lymphoid compartments

studied. Correlated with this decrease in cell count was an increase in number of apoptotic cells in each

organ. Stimulation to alpha adrenergic receptors in rats produces changes in adhesion molecule expres-

sion that leads to leukocyte redistribution. Concurrently, many lymphocytes undergo apoptosis, resulting

in a profound drop in cell numbers in the various lymphoid organs.

U. Stockhorst1, S. Hausmann1, J. Wolf2, P. Enck3, G. Hall4, S. Klosterhalfen1

1Institute of Medical Psychology, and 2Institute of Physiological Psychology, University of Düsseldorf,

Düsseldorf, Germany; 3Department of General Surgery, University of Tübingen, Tübingen, Germany;

4Department of Psychology, University of York, York, England

78. Rotation-induced motion sickness in healthy humans: Subjective symptoms, cortisol, and TNF

Sickness behaviour and related immunological and endocrine changes are quite well examined in ani-

mals, but studies with humans are rare. In this study we used rotation to induce motion sickness in

healthy humans and analysed its subjective (symptoms), endocrine (cortisol), and immunological (tumor

necrosis factor [TNF]-alpha) correlates. Fifty healthy subjects (25 females, 25 males; M = 25.8 years;

non-smokers) were rotated in the vertical axis (120 degrees/sec, 1 minute per rotation) with their eyes

closed and their heads bent up and down. There was a maximum of 5 rotations separated by 1-minute

breaks. Subjects could determine the end of rotation. Measures at baseline, immediately after, 15 min-

utes (min+15) and 30 minutes (min+30) after the individually last rotation included self-reported symp-

toms (among them ńausea´ and úrge to vomit´), as well as cortisol and TNF measured in saliva. In-

tragroup analyses revealed an increase of symptoms from baseline to all subsequent measurement

points, reaching its maximum immediately after rotation 1 ( p < .001). In TNF, we also found an increase

culminating after the individually last rotation (p < .01). Cortisol increased by min+15 and reached its

maximum with a temporal delay in min +30 (p < .001). The intensity of the symptoms ńausea´ and úrge

to vomit´ was positively correlated (Spearman´s rank correlations) with TNF. The results contain first hints

that TNF might be an interesting indicator of nausea and related symptoms. We will examine TNF in other

nausea-inducing interventions, e.g. chemotherapy in cancer patients. (Supported by the WellcomeTrust).

R.H. Straub, H. Herfarth, W. Falk, J. Schölmerich, T. Andus

Department of Internal Medicine I, University Hospital Regensburg, Germany

79. Uncoupling of the sympathetic nervous system and the hypothalamic-pituitary-adrenal-axis in inflammatory bowel disease

In chronic inflammatory diseases such as Crohn's disease (CD) and ulcerative colitis (UC), tumor necro-

sis factor (TNF) or interleukin (IL)-6 stimulate the hypothalamus, which activates the two main stress

axes: the hypothalamus-autonomic nervous system (HANS) axis and the hypothalamic-pituitary-adrenal

(HPA) axis. The two axes are expected to be stimulated in a parallel fashion to dampen inflammation in

the gut. The study was initiated in order to investigate the parallelism of the two axes. We measured a

typical marker of the HANS axis (neuropeptide Y, NPY= neurotransmitter of the sympathetic nerve termi-

nal) and of the HPA axis (cortisol) in samples from control subjects, patients with CD, and patients with

UC together with serum TNF and IL-6. Plasma NPY was elevated in patients with IBD compared to con-

trol subjects (p<0.001, independent of gender, disease activity, prior therapy, and serum IL-6 or TNF).

Plasma NPY was positively correlated with serum cortisol in control subjects (R= 0.259,= p=0.026) which

is a sign for the parallel activation of the two axes in the normal situation. However, serum cortisol was

not correlated with plasma NPY in CD or UC patients without prior prednisolone therapy. In active CD or

UC, inclusion of patients with and without prior prednisolone revealed a negative correlation between

serum cortisol and plasma NPY (CD: R= -0.285, p<0.05=; UC: R= -0.510, p<0.01). In contrast, in view of

continuous inflammation serum cortisol levels were inadequately low in the groups of IBD patients as

compared to control subjects. This study demonstrates that the two stress axes seem to act in a parallel

fashion in control subjects but are uncoupled in IBD patients without prior corticosteroid therapy. This may

be a relevant factor for the chronicity of the diseases. Supported by the Kompetenznetz “Darmerkrankun-

gen”, BMBF.

R.H. Straub1, J.M. Kittner2, C. Heijnen3, M. Schedlowski4, R. E. Schmidt2, R. Jacobs2

1Department of Internal Medicine I, University Hospital Regensburg, Germany, 2Department of Clinical

Immunology, Hannover Medical School, Hannover, Germany, 3Department of Pediatric Immunology,

University Medical Centre of Utrecht, The Netherlands, 4Institute of Medical Psychology, University of

Essen, Essen, Germany

80. Infusion of adrenaline decreases serum levels of cortisol and 17beta-hydroxyprogesterone in patients with rheumatoid arthritis but not in healthy subjects

This study was initiated in order to investigate the pituitary and adrenal hormone response after an intra-

venous adrenaline challenge in patients with rheumatoid arthritis (RA) and healthy subjects (HS). Fifteen

untreated female RA patients (age: 51.5 ± 3.2 yr) and 7 female HS (48.0 ± 4.3 yr) were infused with

adrenaline (0.05 microgram / kg / min) for approx. 20 min. Plasma levels of adrenocorticotropic hormone

(ACTH), and serum levels of cortisol, 17beta-hydroxyprogesterone (17OHP), and dehydro-

epiandrosterone sulphate (DHEAS) were analysed at baseline and shortly after cessation of adrenaline

infusion (20 min). At baseline and after adrenaline infusion, serum levels of cortisol (p=0.045) and 17OHP

(p=0.021) were higher in healthy subjects as compared to RA patients. In contrast, at baseline and after

adrenaline infusion, plasma levels of ACTH and serum levels of DHEAS were similar in HS as compared

to RA patients. After adrenaline infusion, only patients with RA as compared to HS demonstrated a sig-

nificant decrease of serum cortisol (p=0.026) and serum 17OHP (p=0.026). Plasma levels of ACTH

(p=0.073) and serum levels of DHEAS (p=0.055) tended to decrease. This study demonstrates that se-

rum cortisol and 17OHP (cortisol precursor) were lower in patients with RA as compared to HS despite

similar ACTH levels. Furthermore, simulation of an adrenomedullary stress response by adrenaline infu-

sion decreased serum cortisol and 17OHP only in RA patients but not in HS. Such a response may play

an unfavourable role during a typical stress reaction in RA patients which may lead to a more proinflam-

matory situation. Supported by the Volkswagen-Foundation (I/723032/031).

R.H. Straub1, L. Paimela2, R. Peltomaa3, J. Schölmerich1, M. Leirisalo-Repo3

1Department of Internal Medicine I, University Hospital Regensburg, Regensburg, Germany, 2Orton

Hospital, Invalid Foundation, Helsinki, Finland 3Department of Medicine, Division of Rheumatology, Hel-

sinki University Central Hospital, Helsinki, Finland

81. Inadequately low serum levels of steroid hormones in relation to IL-6 and TNF in untreated patients with early rheumatoid arthritis and reactive arthri-tis

In order to compare levels of steroid hormones in relation to cytokines and to study levels of cortisol or

dehydroepiandrosterone (DHEA) in relation to other adrenal hormones in untreated patients with early

rheumatoid arthritis (RA) and reactive arthritis (ReA) as compared to healthy controls. In a retrospective

study with 34 patients with RA, 46 with ReA, and 112 healthy subjects (HS), we measured serum levels

of interleukin (IL)-6, tumor necrosis factor (TNF), adrenocorticotropic hormone (ACTH), cortisol, 17OH-

progesterone, androstenedione, DHEA, and DHEA sulphate (DHEAS). Patients with RA had higher se-

rum levels of IL-6, TNF, cortisol, and DHEA as compared to ReA and HS but no difference was noticed

with respect to ACTH and DHEAS. However, in RA and ReA as compared to HS, levels of ACTH, corti-

sol, androstenedione, DHEAS, and 17OH- progesterone were markedly lower in relation to IL-6 and TNF.

Furthermore, the number of swollen joints inversely correlated with the ratio of serum cortisol / serum IL-6

in RA (R= -0.582, p=0.001) and, to a lesser extent, in ReA (R= -0.417, p=0.011). In RA patients, mean

grip strength of both hands was positively correlated with the ratio of serum cortisol / serum IL-6 (R=

0.472, p=0.010) and serum DHEAS / serum IL-6 (R= 0.658, p<0.001). Furthermore, in these untreated

patients with RA and ReA, there was a relative decrease of hormone secretion of 17OH-progesterone,

androstenedione, and DHEAS in relation to DHEA and cortisol. This indicates a relative predominance of

the non-sulfated DHEA and cortisol in relation to all other measured adrenal steroid hormones in these

early inflammatory diseases. This study indicates that ACTH and cortisol are relatively low in relation to

IL-6 and TNF in untreated patients with early RA and ReA as compared to HS. It further demonstrates

that there is a relative increase of DHEA and cortisol in relation to other adrenal hormones such as

DHEAS. This study emphasizes that adrenal steroid secretion is inadequately low in relation to inflamma-

tion. Although changes of hormone levels are similar in RA and ReA, alteration of the steroidogenesis is

more pronounced in patients with RA as compared to ReA. Supported by the institutions.

R.K. Stumm1, J. Rummel1, C. Culmsee2, V. Junker2, J. Krieglstein2, V. Höllt1, S. Schulz1

1Institute of Pharmacology and Toxicology, Otto-von-Guericke-University Magdeburg, 2Institute of Phar-

macology and Toxicology, Philipps University Marburg

82. Neuronal- and endothelial-specific SDF-1 isoforms differentially modulate CXCR4-dependent neurotransmission and cerebral infiltration after focal cerebral ischaemia and peripheral immune-stimulation by LPS

The alpha-chemokine stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4 are key modulators

of immune function and neuronal development. Here, we examined the cellular expression patterns of

CXCR4 and SDF-1 isoforms after mild peripheral immune-stimulation by LPS and permanent focal cere-

bral ischaemia in mice using in-situ-hybridisation (ISH), confocal microscopy and co-labelling techniques.

SDF-1 transcripts containing the SDF-1beta exon (SDF-1beta mRNAs) were expressed by von Wille-

brand Factor-immunoreactive endothelial cells but not by neurons or microglia. LPS-stimulation and focal

ischaemia dramatically decreased endothelial SDF-1beta mRNA expression throughout the forebrain. In

addition, constitutive expression of neuronal SDF-1 isoforms was detected in the cortex, dentate-gyrus

granule cells and basolateral amygdala in a manner partially overlapping with neuronal CXCR4 expres-

sion. CXCR4-immunoreactivity exhibited somatodendritic and axonal subcellular targeting. Presumed

non-neuronal CXCR4 expression was seen in the ventricular ependymal layer but not in quiescent or

ischaemia-activated glia. Expression levels of neuronal SDF-1 and CXCR4 were unchanged after LPS-

stimulation but transiently changed after ischaemia in the non-lesioned cortex. At the border of the non-

lesioned and infarcted areas 2d after ischaemia a massive increase in SDF-1beta mRNAs was induced in

endothelial cells. This upregulation was in close spatio-temporal overlap with infiltrates of CXCR4 ex-

pressing cells. Within the focus after 4d and 8d an increased SDF-1 and CXCR4 mRNA expression was

observed which may be related to re-organisation of the infarct. Thus, isoform-specific changes of SDF-1

expression may differentially modulate neuronal communication and interactions between the cere-

brovascular endothelium and peripheral immunocytes in both the LPS-stimulated unlesioned and the

ischaemia - lesioned brain.

L. Torner1 , A. Wigger2 , S. Krömer2 , I.D. Neumann1

1Institute of Zoology, University of Regensburg, Regensburg, Germany, 2Max Planck Institute of Psychia-

try, Munich, Germany

83. Brain neuropeptides oxytocin and prolactin: involvement in neuroendocrine and emotional stress coping strategies

Oxytocin and prolactin are reproduction-related (neuro)peptides which were originally seen in context of

maternal behaviour. However, in response to psychological (social defeat, maternal defeat) and physical

(swim) stressors, oxytocin is not only released into blood, but also within hypothalamic and limbic regions

of the rat brain, respectively. Such locally released oxytocin is involved in the regulation of the re/activity

of the hypothalamo-pituitary-adrenal (HPA) axis independent of gender, but dependent on the reproduc-

tive state of female rats. Thus, icv administration of an oxytocin receptor antagonist (OXT-A) reveals a

tonic inhibitory effect upon basal as well as stress-induced ACTH and corticosterone secretion in male

and virgin female, but not pregnant and lactating, rats. Using reversed microdialysis for local OXT-A ad-

ministration, the paraventricular nucleus, the septum and the amygdala could be shown as sites of OXT

action. In contrast, a similar inhibitory effect of brain oxytocin could not be found in mice. In rats, brain

oxytocin acts as an endogenous anxiolytics in the peripartum period. Similar to oxytocin, the brain prolac-

tin system inhibits basal and stress-induced HPA axis activity in female rats, and acts as an effective en-

dogenous anxiolytics. Because a selective prolactin receptor (PRL-R) antagonist is not available, an-

tisense targeting improved by molecular modelling of the targeted PRL-R mRNA had to be employed in

order to downregulate PRL-R synthesis. As both oxytocin and prolactin can be considered anti-stress

factors of the brain integrating neuroendocrine and behavioural functions related to stress coping, our

future studies will search for possible positive effects on immunological parameters. Supported by DFG

and DAAD.

M. Ulmschneider, M. Riedel, B. Brandstätter, C. Scheppach, R. Gruber, N. Müller

Psychiatric and Psychotherapeutic Hospital, Ludwig-Maximilian University, Munich

84. Immunological Effects of Celecoxib and Risperidone in the Course of a Comparative Therapy Study

In the last years there have been several publications indicating an immunological pathogenesis of

schizophrenia in at least a subgroup of patients. Also the existence of immunomodulatory properties of

neuroleptics has been described. We performed a trial with the selective cycloxygenase-2 inhibitor cele-

coxib to evaluate the influence of immunomodulatory effect on therapeutic efficacy and immunological

parameters in patients with schizophrenia. Our study was performed as a double-blind, placebo-

controlled, parallel-group evaluation of risperidone plus celecoxib versus risperidone plus placebo over a

course of five weeks. Both groups showed significant improvement on the PANSS total score and all sub-

scores during the five weeks of treatment with risperidone. The celecoxib add-on therapy group had a

significantly better outcome over the study period regarding the PANSS total score. Looking at the immu-

nological parameters a significant decrease of B-lymphocytes (CD19+) and CD5+B-lymphocytes could be

detected in both groups. Only the celecoxib group showed a significant positive correlation of B-

lymphocytes (CD19+) decrease and an improvement on the PANSS negative score after week 4 and 5.

The subgroup of CD5+B-lymphocytes showed this significant correlation after week 4. No significant addi-

tional effect of celecoxib was observed, however the correlation of the improvement on the PANSS nega-

tive score with the decrease of B-lymphocytes may be attributed to the immunomodulatory properties of

celecoxib.

H. Vedder, S. Lanquillon, J.-C. Krieg

Department of Psychiatry and Psychotherapy, Philipps- University of Marburg, Marburg/Lahn, Germany

85. Regulation of tumor-necrosis factor-alpha production by glucocorticoids in major depression

Steroid modulation of the production of the cytokine tumor- necrosis factor-a (TNF) was studied in normal

controls and in patients with major depressive disorder. Patients were classified as responders and non-

responders with regard to the psychopathological outcome (Hamilton and Montgomery-Asberg depres-

sion scales [MADRS]) after a 6-week antidepressant treatment. Glucocorticoid-modulated production of

TNF was examined under basal conditions and after stimulation with phytohaemagglutinin. In addition to

two concentrations (25, 125 ng/ml) of Cortisol (CORT), the physiological corticosteroid of the human,

dexamethasone (DEX) (5, 25 ng/ml) was used for the in vitro modulation of cytokine production. Under

basal conditions, both steroids induced increases in TNF levels in both patients' subgroups at the pre-

treatment time point, whereas the stimulated production of TNF was significantly decreased by CORT in

the non-responder subgroup at the pre- treatment condition and in both non-responders and responders

at the post-treatment condition. Incubation with the different concentrations of DEX resulted in significant

decreases under all stimulated conditions. Correlation analysis revealed a significant relationship be-

tween the CORT125-modulated TNF production under basal conditions and the psychopathology

(MADRS). Thereby, our data point to differential effects of CORT and DEX on the production of TNF and

a significant relationship between the modulation of TNF production by the physiological corticosteroid

CORT - but not the synthetic glucocorticoid DEX - and the psychopathology of depressive patients.

C. Vogel1, R. Mössner2, T. Heinemann2, P. Riederer2, M. Gerlach2, D.L. Murphy3, K.-P. Lesch2, C.

Sommer1

1Department of Neurology and 2Psychiatry, University of Würzburg, Germany, 3Laboratory of Clinical

Science, NIMH-NIH, Bethesda, USA

86. Changes of serotonin tissue concentration modulate neuropathic pain: A study in serotonin transporter-deficient mice

Serotonin (5-HT) being an important modulator of pain at various levels of the nervous system, we hy-

pothesised that mice deficient of the serotonin transporter (5-HTT) would display altered pain-related

behaviour after nerve injury. We used C57Bl/6-mice deficient for the 5-HTT (5-HTT-/- mice) and com-

pared their pain-related behaviour after a chronic constrictive sciatic nerve injury (CCI) with that of wild-

type littermates. Thermal hyperalgesia was measured according to Hargreaves, mechanical allodynia

with von-Frey-hairs, cold allodynia was assessed using an acetone stimulus. Tissue of footpads, sciatic

nerves, and the L4/5 spinal cord was harvested on day 28. 5-HT and 5-hydroxyindoleacetic acid (5-HIAA)

were determined by high performance liquid chromatography (HPLC). In contrast to wild-type mice, 5-

HTT-/- mice did not develop thermal hyperalgesia after CCI. Both genotypes developed reductions in

mechanical thresholds on the nerve-injured side, but in 5-HTT-/- mice these were present bilaterally. Re-

actions to acetone were without differences between groups. HPLC revealed a significant increase in 5-

HT levels in the injured nerve in both genotypes, while 5-HTT-/- mice had significantly lower overall levels

than wild-type mice in the sciatic nerve and in the spinal cord. Thus, in 5-HTT-/- mice, a reduction of 5-HT

in the peripheral nerve is associated with the absence of thermal hyperalgesia after nerve lesion while

allodynia, a centrally mediated pain-related behaviour, is increased. We conclude that 5-HT contributes to

thermal hyperalgesia after CCI by peripheral sensitisation and that spinal 5-HT prevents contralateral

spread of pain in normal animals.

A. Volz1, A. Ehlers1, S. Beck 2, J. Younger2, A. Ziegler1

1Institute for Immungenetics, University Clinic Charité, Humboldt-University Berlin, Germany; 2The

Sanger Centre, Wellcome Trust Genome Campus, Hinxton, UK

87. Transcripts of HLA-linked olfactory receptor genes exhibit premature polyadenylation, exon sharing and long-distance alternative splicing

We have previously identified 39 olfactory receptor (OR) genes located at the telomeric end of the ex-

tended human MHC. In this study, weak expression for various OR in testis, heart, kidney and lung (e.g.,

hs6M1-16,-3,-6,-18,-20,-21,-27, and hs17M1-20 in testis) could be demonstrated with multi-tissue RNA

blots. In order to analyse the regulation of MHC-linked OR genes, their genomic organisation was deter-

mined using transcripts from testis, kidney and lung. While transcripts of hs6M1-3,-6, and -20 comprised

no untranslated exons, an entirely different situation was seen with hs6M1-21,-27, and -18 (OR genes

with identical transcriptional orientation in the depicted order): testicular transcripts occurred in several

different splice variants, and the most distant 5' exon of hs6M1-21 was found more than 100 kb upstream

of the coding exon, indicating that this OR gene spans an exceptionally large genomic region. Interest-

ingly, this region contains five additional OR genes, whereas hs6M1-21 even shares the most 5' exons

with hs6M1-27 and -18. Therefore, hs6M1-18, -21, and -27 are likely to share also a common promoter

which might additionally regulate hs6M1-16 whose most 5' exon was found only 80 bp upstream, with

opposite transcriptional orientation. The analysis of the coding and the 3' untranslated regions resulted in

additional 3' exons, intra-coding region splicing and premature polyadenylation. In conclusion, these most

unusual properties might be relevant for the regulation of genes belonging to the largest known gene

family. The possible relevance of our findings for selection processes in reproduction will be discussed as

well. Supported by the Volkswagen Foundation (I/75 196) and the Wellcome Trust.

G. S. von Banchet1, O. Schmitt1, R. Bräuer2, H.-G. Schaible1

1Institute of Physiology and 2Pathology, Friedrich-Schiller University of Jena, Germany

88. Regulation of the expression of peptide receptors on cultured DRG neurones In previous studies in the rat we have shown that the expression of neurokinin 1 (NK1) bradykinin (B2)

and somatostatin 4 (sst4) receptors in the lumbar DRG neurones is vastly enhanced at different states of

antigen-induced arthritis (AIA) in one knee joint. The aim of this study was to identify mediators that may

be responsible for the upregulation of the NK1 and B2 receptors in DRG neurones. DRG neurones from

normal rats were cultured overnight in Ham`s F12. The following two days every 2 hours PGE2, TNF,

substance P or KCl were added to the cultures. In further experiments the neurones were cultured without

NGF. BK- and SP-gold or an anti-NK1 receptor antibody were used to determine the expression of the

receptors. The expression of the B2 receptor can be influenced by NGF (via the p75NTR). The expres-

sion of the NK1 receptor was upregulated by PGE2 and by SP in low concentrations but not by NGF.

None of the other tested substances influenced the expression of those receptors. Monoarticular AIA

leads to a marked bilateral upregulation of B2, NK1 and SST4 receptors in DRG neurones. The present

data show that the upregulation of the B2 and the NK1 receptor is triggered by different mediators and

therefore presumably by different regulatory pathways. Supported by the DFG (Scha 404/9-2).

S. von Hörsten1, S. Bedoui1, H.P. Nguyen1, H. Nave1, A. Kask2, R. Pabst1

1Department of Functional and Applied Anatomy, Medical School of Hannover, Germany, 2Department of

Pharmacology, University of Tartu, Estonia

89. Models of stress protection by NPY: NPY/CRH interactions in anxiety regula-tion and NPY/epinephrine interactions in adrenergic leukocytosis

Several physiological effects of NPY are opposite compared to classical stress hormones such as CRH

and epinephrine (EPI). It is unknown whether NPY acts also as a functional antagonist of stress-induced

responses such as anxiety or leukocytosis in vivo. The present studies examined the modulation of CRH-

induced anxiety and epinephrine-induced leukocyte mobilisation by NPY pre-treatment in vivo. Bilateral

microinjections of NPY and CRH into the lateral and the medial septum of rats followed by a social inter-

action test of anxiety were used to investigate the interaction of these two neuropeptide in a limbic struc-

ture. Combined intravenous application of NPY with EPI in freely behaving rats and FACS analysis were

used to monitor co-transmission of NPY and EPI on leukocyte mobilisation. Microinjections of NPY dose-

dependently decreased anxiety, in contrast to the opposite effects of CRH. The anxiogenic-like effect of

CRH was reversed by NPY pre-treatment demonstrating a differential role of these peptides in anxiety

regulation within the lateral septum. Intravenous application of NPY followed by EPI dose-dependently

facilitated, intensified, or inhibited adrenergic leukocytosis. Pre-treatment using a NPY Y1R agonist inhib-

ited the EPI effect while an Y5R agonist potentiated leukocyte mobilisation by EPI. NPY specifically an-

tagonises CRH-induced anxiety in the brain and adrenergic leukocytosis in the blood. In general, these

observations demonstrate that neuropeptide-interactions and co-transmission determine to a large extend

the stress-induced behavioural, neuroendocrine, and immunological responses in vivo.

Supported by the DFG/GRK99 (AK) and the VW-Foundation (I/75169) (SvH, HN, RP).

D. von Holst, F. Uhl and W. Stöhr


90. Love in tree shrews (Tupaia belangeri): proximate causes and physiological consequences

Tree shrews (order Scandentia) are small diurnal mammals distributed throughout Southeast Asia. In the

wild, they usually live in pairs in territories. Contrary to expectations, housing of a male and a female to-

gether results mostly in an "inharmonious pairing" characterised by social tension, as evident from occa-

sional fights, avoidance behaviour, and increased heart rates. In about 20% of all pairings, however, con-

tact between an unfamiliar male and female is characterised from the outset by amicable behaviour: Both

individuals groom and "greet" each other frequently with long bouts of mouth licking. They eat and rest

together and always sleep in the same nestbox. Furthermore their heart rates as well as the serum levels

of cortisol and epinephrine are reduced and several indicators of immune function show improvement; the

opposite is found in inharmonious pairings. After separation of the pairs, the physiological data of the

individuals not only return to initial levels, but change in opposite directions: Compared to the situation

before the pairing the welfare of former harmonious pair is diminished, while it is improved in inharmoni-

ous pairs. The quality of a pairing is based on individual preferences of the females for the scent of the

males: A male that has been fiercely rejected by one female may be accepted immediately by another

female. If females are given the opportunity to choose between several males, there is incest avoidance;

furthermore, sisters show a preference for the same males which indicates a genetic base (involvement

of the MHC-complex) for the females’ choice. Supported by the Volkswagen-Foundation (I/67034).

M. Wahle, S. Kölker2, H. Häntzschel, A. Krause3, C.G.O. Baerwald

Medical Clinic and Polyclinic IV, University of Leipzig, Germany, 2University Childrens Hospital Heidel-

berg, Germany 3Medical Clinic of Rheumatology and Clinical Immunology, University Clinic Charité,

Humboldt-University Berlin, Germany

91. Catecholamine-induced apoptosis of B lymphocytes is reduced in patients with rheumatoid arthritis

There is a close interaction between the autonomic nervous system and the immune system. The ex-

pression from beta2-adrenergic receptors (beta2-R) on lymphocytes is reduced in patients with rheuma-

toid arthritis (RA). We investigated in vitro the effect of beta2-R stimulation on the apoptosis of B lympho-

cytes in patients with RA compared to healthy controls. CD19+ cells (B-Lymphocytes) were isolated from

PBMC of RA patients and healthy controls via magnetic cell separation (MACS) Cells were stimulated

with anti-IgM-mAb, and con-incubated with isoprenaline (10(-9) – 10(-5) M) or medium for 20 h. Apoptosis

was detected via Annexin-V binding and Propidium – Iodide. Caspase-3-activation was measured using

an anti-active caspase-3 antibody. Stimulation of the beta2-R with 10(-5) M isoprenaline (Iso) significantly

increased the frequency of apoptotic cells in healthy controls (medium: 21.2 ± 1.6%, Iso: 34.6 ± 4.4%,

p<0,02). An activation of caspase-3 was simultaneously observed. In contrast, RA patients only showed a

small increase of the number of apoptotic cells (medium: 21.9±1.2%, Iso: 27.9±3.3%, p>0.05). The rela-

tive increase in apoptotic cells in healthy controls (59.3 ± 10.1%) was significantly higher than in RA pa-

tients (21.8 ± 8.9%, p=0.02). Our data demonstrate that the reduction of beta2-R expression on B-

lymphocytes in patients with RA has also functional consequences, whereby the role of the reduced

apoptosis after beta2-R in the pathogenesis of RA requires clarification in future experiments.

R. Wank

Institute for Immunology, Ludwig-Maximilians University Munich, Munich, Germany

92. Substantial improvements in three patients with bipolar, schizophrenic or autistic disorder by longterm low-dosed adoptive immunotherapy

Bidirectional communications between immune cells and the central nervous system have been well rec-

ognised (1), also autoimmune features of schizophrenia. Polarisation of T helper (Th) cells by chronic

infections can influence immune cell secretion of neurotrophins BDNF and NT-3 (2), which are important

for neuronal functions. CD3-activation of T cells in mononuclear cell cultures from peripheral blood can

"clean" or lyse chronically infected cells and reactivate downregulated immune cells. Weekly reinjections

of 5-30 million of such activated autologous immune cells showed substantial improvements of social

competence and other symptoms in at least five years-diseased patients with bipolar (72y), schizophrenic

(32y) and autistic (9y) disorder. Immune and neuronal cytokine levels of activated or non-activated mono-

nuclear cells appeared normal, but IgG and IgA antibody titres against chlamydia pneumonia were re-

markably high in the three patients. The schizophrenic patient had also a recurring EBV infection. Re-

peated azythromycine treatment in the latter showed antipsychotic but limited effects, also on antibody

titers.

1. HO, Sorkin E. Network of immune-neuroendocrine interaction. Clin Exp Immunol 27:1-12 (1977).

2. 2. Besser M and Wank R. Clonally restricted production of the neurotrophins BDNF and NT-3 mRNS

by human immune cells and Th1/Th2-polarized expression of their receptors. J Immunol162:6303-6

(1998).

E. Weihe1, C. Depboylu1, L.E. Eiden2

1Institute of Anatomy and Cell Biology, Philipps University, Marburg, Germany, 2Laboratory of Cellular

and Molecular Regulation, NIMH, Bethesda, USA

93. Responses of neuronal, glial, and endothelial cell compartments of the brain to lentiviral infection an effects of antiretroviral treatment: A new conceptu-alisation of Neuro-AIDS

Lentiviral infection of the primate brain results in complex molecular and cellular events. Some of these

are related to the role of the brain as a viral reservoir in AIDS, and some to the dementia associated with

infection (neuro-AIDS, or AIDS dementia complex. We have previously established that the simian immu-

nodeficiency virus (SIV)-infected rhesus monkey is an appropriate primate model for the study of human

AIDS dementia complex (Murray et al., Science 255, 1246-1249, 1992). In the decade since, we have

characterised the complex molecular and cellular events occurring during the course of SIV infection in

the rhesus monkey. Here, we summarise these findings and propose that neuro-AIDS may be caused not

by overt damage to neurons and/or glia, but by relatively subtle neurochemical changes in both neurons

and glial cells driven by endogenous brain inflammatory responses to viral infection that become irre-

versibly imprinted within these cells in the course of disease. We compared the spatio-temporal relation-

ship of microgliosis / astrogliosis, macrophage infiltration, occurrence of multinucleated giant cells and

cellular proliferation, changes in endothelial cell protein expression, and neurodegenerative changes in-

cluding dendritic and synapse loss that occurred in the brain during progression to AIDS after SIV infec-

tion. Rhesus monkeys with symptoms of AIDS were treated with a CNS-permeating antiretroviral agent,

6-Cl-ddG. Reversal of virus-induced alterations in cellular components of neurons, glial, endothelium,

microglia and invading macrophages were then correlated with decreased virus burden following upon

treatment with 6-Cl-ddG. Major molecular candidate pathogenicity factors implicated in neuroinflammatory

and neurodegenerative brain disease that were followed included i) complement C1q, an important effec-

tor arm of immune defence and complement activation and a putative neuronal and glial factor in neu-

rodegeneration in the course of Alzheimer’s disease and stroke; ii) indoleamine 2,3-dioxygenase

(IDO),the enzyme responsible for generating the neurotoxic quinolinic acid; iii) inducible nitric oxide syn-

thase, responsible for the generation of the putative cytotoxic NO acting as a free radical to generate

peroxynitrates/nitrotyrosine causing oxidative stress and cytotoxicity iv) cyclooxygenases generating

prostaglandins potentially contributory to neuronal hyperexcitability and neurotoxicity; and v) the

chemokine fractalkine (CX3C) and its receptor CX3CR1 (V28) which are involved in regulating chemo-

taxis during retroviral infection and potentially autoimmune disease, and in activating microglia during

lesion-induced neurodegeneration. Using these markers and their patterns of expression in neurons, glia,

microglia, macrophages and endothelial cells, we have discovered distinct virally-induced changes in

each of these cellular compartments of the brain during lentiviral infection. These changes fall rather dis-

tinctly into two classes on the basis of their reversibility by antiretroviral treatment which dramatically de-

creases virus burden in the brain. On this basis, a conceptual framework for the molecular interactions

between neurons, glia and inflammatory cells of the brain that lead to both reversible and irreversible

alterations during and following immunodeficiency virus infection has been constructed. A hypothesis

arising from these investigations is that AIDS dementia complex may reflect initially reversible neuro-

chemical alterations that become 'imprinted', and resistant to alteration with antiretroviral treatments, at

some point in the course of brain lentiviral infection and virus-induced brain inflammatory responses.

Supported by VW-Foundation.

A. Wölfler1,2, K. Schauenstein2, P.M. Liebmann2

1Department of Medicine, Division of Haematology and 2Department of Pathophysiology, University of

Graz, Austria

94. Pro- and antioxidant effects of melatonin in lymphoid cells

Melatonin (MEL), the main endocrine product of the pineal gland, has been claimed to exhibit distinct

antioxidant features in vitro as well as in vivo, which has been considered to be part of its physiological as

well as pharmacological effects. Since reactive oxygen species (ROS) are known to influence a variety of

biologic processes and to constitute a major signal for programmed cell death in normal and tumor cells,

we were interested whether MEL may interfere with intracellular ROS formation in lymphoid cells, e.g. in

peripheral blood lymphocytes (PBL) as well as in Jurkat cells, a human leukaemic T-lymphoblastic cell

line. While in human PBL pharmacological concentrations of MEL exhibited antioxidant properties against

t-butylated-hydroperoxide- and diamide-induced ROS-formation and subsequently decreased the per-

centage of cells undergoing ROS-induced cell death, in resting Jurkat cells MEL exhibited prooxidant

activity, as was shown by increased intracellular oxidation of ROS-sensitive fluorescent dyes. Further-

more, enhanced ROS formation by MEL led to a promotion of fas- and ROS-induced cell death in this

leukaemic cell line. The cell death promoting effect of MEL was dependent on intracellular glutathione

concentration as well as inhibited by addition of trolox, a water-soluble analogue of alpha-tocopherol, and

glutathione indicating that this effect of MEL is indeed mediated by formation of intracellular ROS. These

results give evidence that MEL may be a modulator of the cellular redox status in both ways in lymphoid

cells, and is not necessarily protecting against oxidative hazard. Supported by the Austrian Science

Foundation (P9925-med and P12679-med).

E. Zeisberger1, G. Ross2, J. Roth2, T. Hübschle2, R. Gerstberger2, K. Voigt3

1Department of Physiology, Faculty of Biological Sciences, University of South Bohemia, Ceske Budejov-

ice, 2Department of Veterinary Physiology, Faculty of Veterinary Medicine, Justus-Liebig University, Gi-

essen, Germany, 3Institute for Normal und Pathological Physiology, Clinic of the Philipps-University,

Marburg, Germany

95. Are fever-inducing immune signals in part transported to the brain by affer-ent skin nerves?

Fever is induced by peripheral immune signals transported to the brain either by blood stream or by

nerves. To investigate whether cutaneous nerves might participate in the transfer of immune signals to

the brain, guinea pigs were chronically equipped with intra-arterial catheters for repeated blood sampling,

with artificial subcutaneous chambers containing catheters for administration of drugs or repeated sam-

pling of chamber lavage, and with intraabdominally implanted radiotransmitters to monitor body core tem-

perature. Fever was induced by injections of 10 µg/kg lipopolysaccharide (LPS) into the chambers in con-

trol animals or in animals in which the skin surrounding the chamber was anaesthetised by the ropiva-

caine (ROPI, 10 mg/kg administered 30 min prior to LPS). In animals pre-treated with ROPI fever was

reduced to about 60 % of the response in controls. The suppression of fever was not observed when

ROPI was injected into the contralateral skin area so that a systemic effect of ROPI can be excluded.

Whereas LPS and proinflammatory cytokines (TNF, IL-1, IL-6) were measured in high concentrations in

the fluid of chamber lavage by specific assays, only IL-6 spilled over into the circulation in considerable

amounts during the febrile response. Participation of other local mediators (TNF, NO and prostaglandins)

was investigated by addition of specific blockers into the chamber along with the LPS. It is concluded that

afferent neural signals participate in the manifestation of fever in addition to humoral signals mediated by

IL-6, prostaglandins and NO in this model of inflammation.

A.C. Zenclussen, R. Joachim, C. Peiser, J. Niess, B. Klapp, P.C. Arck

Charité, Humboldt University, Berlin, Germany

96. Heme Oxygenase (HO) is downregulated in Stress-triggered Th1-mediated murine abortion

Stress is known to boost the abortion rate in mice by enhancing Th1 cytokines at the fetomaternal inter-

face, thus resulting in thrombosis and haemorrhage. HOs are involved in tissue protection; thus failing

upregulation of HO has been linked to increased necrosis in inflammatory tissues. Here, we investigated

the presence of HOs in mouse placenta and decidua, and their regulation during stress- and Th1-induced

abortion. DBA/2J mated CBA/J female were randomised, exposed to 1) 24 h ultrasound stress at day 5 of

gestation; to 2) injections of recombinant IL-12 or 3) no treatment. At the beginning of late pregnancy the

animals were sacrificed, and the abortion rate registered. The presence of decidual and placental HO-1

and HO-2 was confirmed by immunohistochemistry (IHC); mRNA levels were evaluated using Real Time

PCR. Stress and IL-12-treated mice had significantly higher abortion rates compared to control group.

IHC analysis confirmed the expression of HO in gestational tissue. Real Time PCR revealed that stress or

IL-12 treated group had lower placental and decidual levels of HO-1 and HO-2 mRNA compared to the

control group. Increased Th1 cytokine levels triggered by stress during murine gestation results in low

expression of HOs. It remains to be elucidated whether a lack of upregulation or even a down-regulation

of HOs might be responsible for the rejection phenomenon.

D. Zwingmann, H.O. Besedovsky, A. Kabiersch, H. Rogausch, A. Del Rey

Department of Immunophysiology, Medical Faculty, Philipps-University of Marburg, Germany

97. Bidirectional interactions between splenic sympathetic nerves and the im-mune response during a graft versus host reaction

Splenic sympathetic innervation regulates not only the blood flow in this organ, but also several immune

processes. Although many studies have investigated the effect of noradrenaline (NA) on immune cells,

few reports have focused on in vivo studies. To examine the influence of noradrenergic nerves on differ-

ent lymphoid cell subpopulations and cytokine production during a specific immune response in vivo, we

used as model a systemic graft versus host reaction (GvHR) in animals in which the spleen was dener-

vated locally. Our results show that 10 days after injection of semi-allogeneic lymphoid cells, the percent-

age of activated T lymphocytes (CD5+/IL-2 receptor+ and CD5+/CD134+) is significantly increased as

compared to that of the appropriate controls with a normally innervated spleen. While at this time after

GvH induction the levels of IL-1 beta and IL-2 are within the basal range, those of TNF alpha and IL-10

only slightly increased, and of IFN-gamma slightly decreased in normally innervated spleens, the concen-

tration of these cytokines is significantly increased in denervated spleens. We have also found that during

the GvHR, the amount of NA per 106 nucleated cells is decreased in the spleen of animals with an intact

sympathetic innervation. This result is of particular importance since immune cell functions depend on

microenvironmental conditions. In summary, the results obtained indicate that: 1) NA inhibits antigen spe-

cific immune responses; 2) the reduced NA concentration in the microenvironment of T lymphocytes fa-

vours cell activation; and 3) this results in up regulation of several cytokines. Supported by the Volks-

wagen-Foundation and the SFB 297.

D. Zwingmann, H.O. Besedovsky, A. Kabiersch, H. Rogausch, A. Del Rey

Department of Immunophysiology, Medical Faculty, Philipps-University of Marburg, Germany

98. A new denervation technique to study local noradrenergic influences on specific immune responses in popliteal lymph nodes

We have recently shown that the sympathetic nervous system (SNS) modulates a specific immune re-

sponse in the spleen. To investigate whether comparable local immune-SNS interactions occur in periph-

eral lymph nodes, we developed a technique to denervate the popliteal lymph node (pLN) locally. Under

general anaesthesia, two injections (50µg in 10µl each) of the sympatholytic compound 6-

hydroxydopamine (6-OH-DA) were injected into the fat surrounding the pLN on two consecutive days.

Three days later, the content of noradrenaline (NA) in the pLN was reduced by more than 70%. Since the

reduction of NA in the surrounding fat positively correlates with the degree of denervation of the pLN, the

denervated pLN itself can be used for other studies, for example, for flow cytometry. The reduction of NA

concentration achieved by this method is comparable to that obtained following general chemical sympa-

thectomy, but has the advantage that the sympathetic innervation of other lymphoid organs such as in-

guinal lymph nodes and the spleen, as well as of the foot pad, is not affected. Since the innervation of the

contralateral pLN also remains unaffected, this procedure offers the additional advantage that this struc-

ture can be used as control for local immune responses in the same animal. Furthermore, this minimally

invasive technique reduces the risk of infection and inflammation. Using this method, we found that the

percentage of CD4+ cells is decreased in the denervated pLN 5 days after inducing a local graft vs host

reaction triggered by injection of semi-allogeneic cells into the foot pad. Supported by the Volkswagen-

Foundation and the SFB 297.

(Late Abstract)

C. Depboylu1, T.A. Reinhart2, W.J. Schwaeble1, H. Mitsuya3, D. Rausch4, L.E. Eiden4, E.Weihe1

1Inst. of Anatomy & Cell Biol., Philipps University, Marburg, Germany; 2Dept. of Infectious Diseases &

Microbiology, University of Pittsburgh, USA; 3Div. of Cancer Treatment, NCI and 4Lab. of Cellular & Mo-

lecular Regulation, NIMH, Bethesda, USA

99. Involvement of cerebral C1q biosynthesis in neuro-aids

The complement system as an innate humoral immune defence system is involved in autoimmune, in-

flammatory and degenerative neurological diseases. In this study we investigate the biosynthesis of the

recognition subcomponent of the classical complement activation pathway in the brain of rhesus monkeys

infected with the simian immunodeficiency virus (SIV), a well-documented model for human HIV infection,

AIDS, and HIV-associated neurocognitive dysfunction. Using semiquantitative in-situ hybridisation and

immunohistochemistry a progressive increase in the number of cells expressing C1q was observed

through early to late stages of SIV-infection. Anti-retroviral treatment with a CNS-permeating halogenated

dideoxypurine (6-Cl-ddG) suppressed C1q activation in the brain of monkeys with AIDS. Co-staining re-

vealed cells of mononuclear origin as the only source for C1q synthesis. The increase in the number of

C1q expressing cells was due to activation of gene transcription of its peptide chains, proliferation and

infiltration of C1q positive cells into the brain of monkeys with AIDS. To our knowledge these are the first

data demonstrating microglia/macrophage-lineage-specific regulation of C1q chains and encoding mRNA

in primate brain in different stages of retroviral disease and the relationship of C1q expression to virus

burden during treatment of lentiviral infection with anti-retroviral agents. Supported by Volkswagen-

Foundation.

Contents Abel S 68 Ackenheil M 47, 70 Adamski J 11 Alipour A 1 Allolio B 52 Altmann B 5 Andus T 79 Arck PC 2, 18, 23, 35, 60, 96 Arlt W 52 Arolt V 64, 68 Arzt E 65 Badent B 52 Baerwald CGO 30, 91 Balschun D 3 Beck S 87 Becker G 52 Bedoui S 89 Besedovsky HO 3, 10, 66, 97, 98 Birkmann S 4 Biskup C 61 Böhringer M 19 Bondy B 17 Born J 50 Börner C 46 Brandstätter B 84 Bräuer R 88 Braun A 39, 57, 63 Breivik T 76 Brzoska T 5, 38 Buchberger M 28 Bui NT 64 Bürger K 6 Buske-Kirschbaum A 7, 12 Canbay A 22 Chan A 40 Chiang S 70 Culmsee C 82 Czygan M 20 Dalal M 4, 26 Dawils L 8, 74 Deinzer R 9 Del Rey A 3, 10, 66, 97, 98 Depboylu C 93, 99 Dignass A 23, 60 Dinkel K 11, 55 Dorsch W 28 Ebrecht M 12 Eder S 73 Eggert F 13 Ehlers A 87 Eiden LE 93, 99 Eisel U 14 Elsenbruch S 15, 27, 29 Enck P 78 Engler H 16, 74

Exton MS 22, 61 Falk W 79 Faraji J 1 Fehm HL 50 Fellerhoff B 17 Fest S 18 Fiebich BL 19, 20 Fischbach S 7 Flachenecker P 21 Flentje M 52 Fontaine V 14 Freede S 22 Fuchs D 45, 73 Gerlach M 86 Gerstberger R 95 Giannini E 46 Göbel MU 22, 27 Gold R 40 Goldmann D 23 Gottschalk M 21, 24 Götze J 23 Grabbe S 5 Granrath N 9 Grifka J 56 Grinevich VV 25 Gruber R 84 Haack M 4, 26, 44 Haake P 27 Haen E 28 Haerter K 29 Hall G 78 Hampel H 6 Hanker J 7 Häntzschel H 91 Hartung HP 40 Hauck P 30 Hausmann S 78 Heberling K 27 Heemann U 22 Heijnen CJ 29, 31, 34, 42, 80 Heinemann T 86 Hellhammer DH 7, 12 Henningsen P 55 Herfarth H 79 Herforth A 9 Herklotz J 32 Hettich M 68 Hickfang K 46 Hildebrandt M 2 Hinze-Selch D 44 Höllt V 46, 82 Holsboer F 24 Holtmann G 15 Hoves S 16 Hoyle GW 63 Huber R 45 Hübschle T 95 Hüll M 19, 20

Hummel V 69 Hutzelmeyer H 33 Idel H 9 Jacobs R 31, 34, 37, 42, 80 Jafair Y 1 Jirikovski GF 25 Joachim R 2, 35, 60, 96 Joksimovic L 41 Junker V 82 Kabiersch A 10, 97, 98 Kahl KG 36 Kaiser S 37 Kalden D-H 5 Karanth S 54 Karl T 37 Kask A 89 Kastenbauer S 38 Kavelaars A 29, 31 Kemmler J 45 Kern S 12 Kerzel S 39, 63 Kieseier BC 40 Kirchner H 68 Kirschbaum C 12, 41, 49, 67 Kittner JM 42, 80 Klapp B 18, 96 Klapp BF 2, 35, 60 Klein H 51 Klein M 14 Klosterhalfen S 78 Knopf G 74 Ködel U 38 Kohl C 45 Kölker S 91 Körner C 43, 55 Köthe D 26, 44 Krastel H 75 Kraus J 46 Kraus T 4, 26, 44 Krause A 30, 91 Krieg JC 51, 85 Krieglstein J 82 Krömer S 59, 83 Krönig H 47, 48, 70 Krueger T 27 Kruse N 36 Kümpfel T 21, 24 Kunz SR 49 Kurth S 16, 74 Laich A 45 Lancel DM 72 Lange H 18 Lange T 50 Lanquillon S 51, 85 Laumbacher B 17 Leirisalo-Repo M 81 Lesch K-P 86 Leussink VI 52

Levite M 53 Liebetrau W 4 Liebmann PM 77, 94 Limmroth V 29 Lohrer P 65 Lombardi MS 31 Luger TA 5, 38 Luttmann W 57 Lysson A 15 Marchetti L 14 Marmot M 49 Martin M 34 Mastronardi CA 54 Mayer S 57 McCann SM 54 Meents N 63 Meinck H-M 11, 43, 55, 75 Menger N 75 Mentlein R 37 Meyer-Olsoni D 34 Michel MC 22 Mikeska E 34 Miller LE 56 Mirrezaie A 1 Mitsuya H 99 Möller G 11 Mönnikes H 60 Mössner R 86 Müksch B 19 Müller N 17, 47, 48, 70, 71, 84 Murphy DL 86 Nassenstein C 57 Nast-Kolb D 61 Nave H 89 Neumann H 58 Neumann ID 59, 83 Nguyen HP 89 Niess J 2, 23, 35, 60, 96 Nockher WA 63 Oberbeck R 61 Olshausen B 52 Özcan D 15 Pabst R 37, 76, 89 Pacheco-López G 62 PaezPereda M 65 Paimela L 81 Päth G 39, 63 Pawlak CR 34, 42 Peiser C 96 Peltomaa R 81 Perras B 50 Peschel A 74 Peters M 64, 68 Pfister HW 38 Pfizenmaier K 14 Pitossi F 3 Pollmächer T 4, 26, 44, 72 Prehn JHM 64

Quarcoo D 35, 39 Raap U 39, 63 Rauh W 7 Rausch D 99 Reckemeyer I 51 Reinhart TA 99 Renner U 65 Renz H 39, 57, 63 Richter W 11, 43, 55, 75 Rickert M 11 Rieckmann P 21, 36, 69 Riedel M 47, 48, 70, 71, 84 Riederer P 86 Riegler S 41 Rogausch H 66, 97, 98 Roggero E 10 Rohleder N 41, 67 Rose M 2 Ross G 95 Roth J 95 Rothermundt M 64, 68 Rudolf S 68 Rummel J 82 Ruprecht K 69 Sachser N 37 Schaaf L 65 Schäfer M 10 Schaible H-G 88 Schauenstein K 77, 94 Schedlowski M 15, 22, 27, 29, 31, 34, 42, 61, 62, 80 Scheppach C 71, 84 Schiffold A 30 Schleicher S 20 Schmidt MH 36 Schmidt RE 31, 34, 37, 42, 80 Schmitt O 88 Schmitz D 61 Schneider H 3 Schölmerich J 56, 79, 81 Scholzen T 5 Schröcksnadel H 45 Schuld A 4, 26, 44 Schüler M 61 Schulz S 74, 82 Schwäble WJ 99 Schwarz A 5 Schwarz MJ 47, 48, 70, 71 Schwarz T 5 Shingu K 37 Sokullu S 71 Sölder E 45 Sommer C 86 Sorge S 72 Sperner-Unterweger B 45, 73 Spleiss O 20 Stalla GK 65 Stefanski V 8, 16, 74 Steffen H 75

Steiner JF 22 Stephan M 76 Steptoe A 49 Stevenson JR 77 Stiller-Winkler R 9 Stockhorst U 78 Stöhr W 90 Straub RH 56, 76, 79, 80, 81 Stumm RK 82 Supanz S 77 Tichomirova M 65 Torner L 83 Toyka KV 21, 40 Trebst AE 22 Trenkwalder C 21 Uhl F 90 Ulmschneider M 48, 71, 84 Ungethüm U 30 Vedder H 51, 85 Veit S 52 VirchowJr JC 57 Vogel C 86 Vogel M 17 Voigt K 66, 95 Volz A 87 von Banchet GS 88 von Holst D 33, 90 von Hörsten S 37, 76, 89 Vroon A 29, 31 Wahle M 30, 91 Walch T 45 Wank R 17, 92 Waschul B 9 Weber C 18 Weber F 21, 24 Weihe E 10, 93, 99 Westermann J 32, 77 Widner B 45, 73 Wieland B 43 Wiesmann M 68 Wigger A 59, 83 Wilsenack K 61 Wolf J 41, 67, 78 Wölfler A 94 Xie YF 22 Yassouridis A 24 Yolken R 48, 71 Younger J 87 Yu WH 54 Zeisberger E 95 Zenclussen AC 18, 35, 60, 96 Ziegler A 87 Zuschratter W 3 Zwingmann D 66, 97, 98

PSYCHO NEURO ENDOCRINO IMMUNOLOGY

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