ZIKA VIRUS: Possible immune-prophylaxis and inhibition of biological sequelae and clinical presentation after primary exposure.

DMITRI POPOV. PHD, RADIOBIOLOGY. MD (RUSSIA)

ADVANCED MEDICAL TECHNOLOGY AND SYSTEMS INC. CANADA. INTERVACCINE@GMAIL.COM

ZIKA virus.

Research Proposal: ZIKA VIRUS: Possible immune prophylaxis and inhibition of biological sequelae and clinical presentation after primary exposure.

Dmitri Popov  Full-text · Research Proposal · Jan 2016 Add resources File name: ZIKA.pptx

DOI: 10.13140/RG.2.1.2985.8320

ZIKA virus.

Flaviviridae is a family of viruses. Humans and other mammals serve as natural hosts. They are primarily spread through arthropod vectors (mainly ticks and mosquitoes).

The family gets its name from the Yellow Fever virus, the type virus of Flaviviridae; flavus means yellow in Latin. (Yellow fever in turn was named because of its propensity to cause jaundice in victims.)

 There are currently over 100 species in this family, divided among four genera. Diseases associated with this family include: hepaciviruses: hepatitis; pestiviruses: hemorrhagic syndromes, abortion, fatal mucosal disease; flavivirus: hemorrhagic fever, encephalitis.

ZIKA virus

Genus Flavivirus (type species Yellow fever virus, others include West Nile virus, Dengue Fever and Zika virus)—contains 67 identified human and animal viruses.

Flaviviridae have monopartite, linear, single-stranded RNA genomes of positive polarity, 9.6 to 12.3 kilobase in length. The 5'-termini of flaviviruses carry a methylated nucleotide cap, while other members of this family are uncapped and encode an internal ribosome entry site.

https://en.wikipedia.org/wiki/Flaviviridae

ZIKA virus.

Viral replication is cytoplasmic. Entry into the host cell is achieved by attachment of the viral envelope protein E to host receptors, which mediates clathrin-mediated endocytosis. Replication follows the positive stranded RNA virus replication model. Positive stranded RNA virus transcription is the method of transcription. Translation takes place by viral initiation. The virus exits the host cell by budding. Humans and mammals serve as the natural host. The virus is transmitted via a vector (ticks and mosquitoes)

https://en.wikipedia.org/wiki/Flaviviridae

ZIKA virus.

Major diseases caused by the Flaviviridae family include: Dengue fever Japanese encephalitis Kyasanur Forest disease Murray Valley encephalitis St. Louis encephalitis Tick-borne encephalitis West Nile encephalitis Yellow fever Hepatitis C Virus Infection Zika Virus https://en.wikipedia.org/wiki/Flaviviridae

ZIKA virus.

ZIKA virus.

ZIKA virus.

ZIKA virus.

ZIKA virus.

ZIKA virus.

About 1 in 5 people infected with Zika virus become ill (i.e., develop Zika).

The most common symptoms of Zika are fever, rash, joint pain, or conjunctivitis (red eyes). Other common symptoms include muscle pain and headache. The incubation period (the time from exposure to symptoms) for Zika virus disease is not known, but is likely to be a few days to a week.

The illness is usually mild with symptoms lasting for several days to a week.

Zika virus usually remains in the blood of an infected person for a few days but it can be found longer in some people.

Severe disease requiring hospitalization is uncommon. Deaths are rare.

ZIKA virus.

The symptoms of Zika are similar to those of dengue and chikungunya, diseases spread through the same mosquitoes that transmit Zika.

See your healthcare provider if you develop the symptoms described above and have visited an area where Zika is found.

If you have recently traveled, tell your healthcare provider when and where you traveled.

Your healthcare provider may order blood tests to look for Zika or other similar viruses like dengue or chikungunya.

ZIKA virus.

No vaccine or medications are available to prevent or treat Zika infections. Treat the symptoms:

Get plenty of rest Drink fluids to prevent dehydration Take medicine such as acetaminophen to relieve fever and pain Do not take aspirin and other non-steroidal anti-inflammatory drugs (NSAIDs), like ibuprofen and

naproxen. Aspirin and NSAIDs should be avoided until dengue can be ruled out to reduce the risk of hemorrhage (bleeding). If you are taking medicine for another medical condition, talk to your healthcare provider before taking additional medication.

If you have Zika, avoid mosquito bites for the first week of your illness. During the first week of infection, Zika virus can be found in the blood and passed from an

infected person to another mosquito through mosquito bites. An infected mosquito can then spread the virus to other people.

ZIKA virus.

Complications: ZIKA VIRUS INFECTION COMPLICATED BY GUILLAIN-BARRÉ

SYNDROME – CASE REPORT, FRENCH POLYNESIA, DECEMBER 2013

Oehler E, Watrin L, Larre P, Leparc-Goffart I, Lastère S, Valour F, Baudouin L, Mallet HP, Musso D, Ghawche F. Zika virus infection complicated by Guillain-Barré syndrome – case report, French Polynesia, December 2013. Euro Surveill. 2014;19(9):pii=20720. Article DOI: http://dx.doi.org/10.2807/1560-7917.ES2014.19.9.20720

http://www.eurosurveillance.org/ViewArticle.aspx?ArticleId=20720

ZIKA virus.

Zika fever, considered as an emerging disease of arboviral origin, because of its expanding geographic area, is known as a benign infection usually presenting as an influenza-like illness with cutaneous rash. So far, Zika virus infection has never led to hospitalisation. We describe the first case of Guillain–Barré syndrome (GBS) occurring immediately after a Zika virus infection, during the current Zika and type 1 and 3 dengue fever co-epidemics in French Polynesia.

http://www.eurosurveillance.org/ViewArticle.aspx?ArticleId=20720

ZIKA virus.

Guillain-Barré syndrome (GBS) is a disorder in which the body's immune system attacks part of the peripheral nervous system. The first symptoms of this disorder include varying degrees of weakness or tingling sensations in the legs. In many instances the symmetrical weakness and abnormal sensations spread to the arms and upper body. These symptoms can increase in intensity until certain muscles cannot be used at all and, when severe, the person is almost totally paralyzed. In these cases the disorder is life threatening - potentially interfering with breathing and, at times, with blood pressure or heart rate - and is considered a medical emergency. Such an individual is often put on a ventilator to assist with breathing and is watched closely for problems such as an abnormal heart beat, infections, blood clots, and high or low blood pressure. http://www.ninds.nih.gov/disorders/gbs/detail_gbs.htm

ZIKA virus.

Guillain-Barré syndrome can affect anybody. It can strike at any age and both sexes are equally prone to the disorder. The syndrome is rare, however, afflicting only about one person in 100,000. Usually Guillain-Barré occurs a few days or weeks after the patient has had symptoms of a respiratory or gastrointestinal viral infection. Occasionally surgery will trigger the syndrome. In rare instances vaccinations may increase the risk of GBS.

After the first clinical manifestations of the disease, the symptoms can progress over the course of hours, days, or weeks. Most people reach the stage of greatest weakness within the first 2 weeks after symptoms appear, and by the third week of the illness 90 percent of all patients are at their weakest.

http://www.ninds.nih.gov/disorders/gbs/detail_gbs.htm

ZIKA virus.

What scientists do know is that the body's immune system begins to attack the body itself, causing what is known as an autoimmune disease. Usually the cells of the immune system attack only foreign material and invading organisms.

In Guillain-Barré syndrome, however, the immune system starts to destroy the myelin sheath that surrounds the axons of many peripheral nerves, or even the axons themselves (axons are long, thin extensions of the nerve cells; they carry nerve signals). The myelin sheath surrounding the axon speeds up the transmission of nerve signals and allows the transmission of signals over long distances.

http://www.ninds.nih.gov/disorders/gbs/detail_gbs.htm

ZIKA virus.

In diseases in which the peripheral nerves' myelin sheaths are injured or degraded, the nerves cannot transmit signals efficiently. That is why the muscles begin to lose their ability to respond to the brain's commands, commands that must be carried through the nerve network. The brain also receives fewer sensory signals from the rest of the body, resulting in an inability to feel textures, heat, pain, and other sensations. Alternately, the brain may receive inappropriate signals that result in tingling, "crawling-skin," or painful sensations. Because the signals to and from the arms and legs must travel the longest distances they are most vulnerable to interruption. Therefore, muscle weakness and tingling sensations usually first appear in the hands and feet and progress upwards.

http://www.ninds.nih.gov/disorders/gbs/detail_gbs.htm

ZIKA virus.

When Guillain-Barré is preceded by a viral or bacterial infection, it is possible that the virus has changed the nature of cells in the nervous system so that the immune system treats them as foreign cells. It is also possible that the virus makes the immune system itself less discriminating about what cells it recognizes as its own, allowing some of the immune cells, such as certain kinds of lymphocytes and macrophages, to attack the myelin. Sensitized T lymphocytes cooperate with B lymphocytes to produce antibodies against components of the myelin sheath and may contribute to destruction of the myelin. In two forms of GBS, axons are attacked by antibodies against the bacteria Campylobacter jejuni, which react with proteins of the peripheral nerves.  Acute motor axonal neuropathy is particularly common in Chinese children. http://www.ninds.nih.gov/disorders/gbs/detail_gbs.htm

ZIKA virus.

Scientists are investigating these and other possibilities to find why the immune system goes awry in Guillain-Barré syndrome and other autoimmune diseases. The cause and course of Guillain-Barré syndrome is an active area of neurological investigation, incorporating the cooperative efforts of neurological scientists, immunologists, and virologists.

http://www.ninds.nih.gov/disorders/gbs/detail_gbs.htm

ZIKA virus. Conventional treatment.

There is no known cure for Guillain-Barré syndrome. However, there are therapies that lessen the severity of the illness and accelerate the recovery in most patients. There are also a number of ways to treat the complications of the disease.

Currently, plasma exchange (also called plasmapheresis) and high-dose immunoglobulin therapy are used. Both of them are equally effective, but immunoglobulin is easier to administer. Plasma exchange is a method by which whole blood is removed from the body and processed so that the red and white blood cells are separated from the plasma, or liquid portion of the blood. The blood cells are then returned to the patient without the plasma, which the body quickly replaces. Scientists still don't know exactly why plasma exchange works, but the technique seems to reduce the severity and duration of the Guillain-Barré episode. This may be because plasmapheresis can remove antibodies and other immune cell-derived factors that could contribute to nerve damage.

http://www.ninds.nih.gov/disorders/gbs/detail_gbs.htm

ZIKA virus.

In high-dose immunoglobulin therapy, doctors give intravenous injections of the proteins that, in small quantities, the immune system uses naturally to attack invading organisms. Investigators have found that giving high doses of these immunoglobulins, derived from a pool of thousands of normal donors, to Guillain-Barré patients can lessen the immune attack on the nervous system. Investigators don't know why or how this works, although several hypotheses have been proposed.

http://www.ninds.nih.gov/disorders/gbs/detail_gbs.htm

ZIKA virus.

The use of steroid hormones has also been tried as a way to reduce the severity of Guillain-Barré, but controlled clinical trials have demonstrated that this treatment not only is not effective but may even have a deleterious effect on the disease.

http://www.ninds.nih.gov/disorders/gbs/detail_gbs.htm

ZIKA virus.

The most critical part of the treatment for this syndrome consists of keeping the patient's body functioning during recovery of the nervous system. This can sometimes require placing the patient on mechanical ventilatory assistance, a heart monitor, or other machines that assist body function. The need for this sophisticated machinery is one reason why Guillain-Barré syndrome patients are usually treated in hospitals, often in an intensive care ward. In the hospital, doctors can also look for and treat the many problems that can afflict any paralyzed patient - complications such as pneumonia or bed sores. http://www.ninds.nih.gov/disorders/gbs/detail_gbs.htm

ZIKA virus. Original, experimental method of therapy.

Vascular injury, acutely endothelial damage can lead to vasogenic oedema chronically fibrosis, hyalinization and stenosis can occur with

eventual thrombosis and infarction vascular ectasia, and telangiectasia are also seen frequently,

with capillary telangiectasias.

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

a viral or bacterial infection, it is possible that the virus has changed the nature of cells in the nervous system so that the immune system treats them as foreign cells. It is also possible that the virus makes the immune system itself less discriminating about what cells it recognizes as its own, allowing some of the immune cells, such as certain kinds of lymphocytes and macrophages, to attack the myelin.

oligodendrocytes and white matter damage oligodendrocytes are sensitive to viral infection. loss of white matter accounts for the majority of volume loss

effects on the fibrinolytic enzyme system increase in urokinase plasminogen activator and simultaneous decrease in tissue

plasminogen activator may contribute to cytotoxic oedema and tissue necrosis

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

Perforin and Granzyme Inhibition with monoclonal antibodies.

Protease Inhibition – Protease Inhibitors. MAC - Membrane Attack Complex Inhibition with

monoclonal antibodies.

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

Perforin and Granzyme Inhibition with monoclonal antibodies. http://www.slideshare.net/dlpopov/radiation-cytotoxicity http://

www.slideshare.net/dlpopov/leukocyte-and-lymphocyte-cytotoxicity?related=1

Granzymes are serine proteases that are released by cytoplasmic granules within cytotoxic T cells and natural killer (NK) cells. They induce programmed cell death in the target cell, thus eliminating cells that have become cancerous or are infected with viruses or bacteria. The granzymes also kill bacteria and inhibit viral replication. In NK cells and T cells, the granzymes are packaged in cytotoxic granules with perforin. Other locations that granzymes can be detected are in the rough endoplasmic reticulum, golgi complex, and the trans-golgi reticulum.

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

The contents of the cytotoxic granules function to permit entry of the granzymes into the target cell cytosol. The granules are released into an immune synapse formed with a target cell, where perforin mediates the delivery of the granzymes into endosomes in the target cell, and finally into the target cell cytosol.

Granzymes are identified as being part of the serine esterase family. They are closely related to other immune serine proteases expressed by innate immune cells, such asneutrophil elastase and cathepsin G.

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

Granzyme B activates apoptosis by activate caspases (especially caspase-3), which cleaves many substrates, including caspase-activated DNase to execute cell death. Granzyme B also cleaves the protein Bid, which recruits the proteins Bax and Bak to change the membrane permeability of the mitochondria, causing the release of cytochrome c(which is one of the parts needed to activate caspase-9 via the apoptosome), Smac/Diablo and Omi/HtrA2 (which suppress the inhibitor of apoptosis proteins (IAPs)), among other proteins. Granzyme B also cleaves many of the proteins responsible for apoptosis in the absence of caspase activity. The other granzymes activate cell death by caspase-dependent and caspase-independent mechanisms

ZIKA virus. Original, experimental method of therapy.

In addition to killing their target cells, granzymes can target and kill intracellular pathogens.

Granzymes A and B induce lethal oxidative damage in bacteria by cleaving components of the electron transport chain, while granzyme B cleaves viral proteins to inhibit viral activation and replication. 

The granzymes bind directly to the nucleic acids DNA and RNA; this enhances their cleavage of nucleic acid binding proteins.

ZIKA virus. Original, experimental method of therapy.

More recently, in addition to T lymphocytes, granzymes have been shown to be expressed in other types of immune cells such as dendritic cells, B cells and mast cells. In addition, granzymes may also be expressed in non-immune cells such as keratinocytes, pneumocytes and chondrocytes. As many of these cell types either do not express perforin or do not form immunological synapses, granzyme B is released extracellularly. Extracellular granzyme B can accumulate in the extracellular space in diseases associated with dysregulated or chronic inflammation leading to the degradation of extracellular matrix proteins and impaired tissue healing and remodelling. Extracellular granzyme B has been implicated in the pathogenesis of atherosclerosis, aneurysm, vascular leakage, chronic wound healing, and skin aging.

ZIKA virus. Original, experimental method of therapy.

Excessive Perforin and Granzyme possible play important role in endothelial damage and nervous system damage.

Perforine and Granzyme inhibition by monoclonal antibodies significantly reduce nervous system damage.

ZIKA virus. Original, experimental method of therapy.

Protease Inhibition: http://

www.slideshare.net/dlpopov/radiation-protectionprotease-inhibition Proteases occur in all organisms, from prokaryotes to eukaryotes to 

viruses. These enzymes are involved in a multitude of physiological reactions from simple digestion of food proteins to highly regulated cascades (e.g., the blood-clotting cascade, the complement system, apoptosis pathways, and the invertebrate prophenoloxidase-activating cascade). Proteases can either break specific peptide bonds (limited proteolysis), depending on the amino acid sequence of a protein, or break down a complete peptide to amino acids (unlimited proteolysis). The activity can be a destructive change (abolishing a protein's function or digesting it to its principal components), it can be an activation of a function, or it can be a signal in a signalling pathway.

https://en.wikipedia.org/wiki/Protease

ZIKA virus. Original, experimental method of therapy.

The activity of proteases is inhibited by protease inhibitors. One example of protease inhibitors is the serpin superfamily, which includes alpha 1-antitrypsin, C1-inhibitor,antithrombin, alpha 1-antichymotrypsin, plasminogen activator inhibitor-1, and neuroserpin.

Natural protease inhibitors include the family of lipocalin proteins, which play a role in cell regulation and differentiation. Lipophilic ligands, attached to lipocalin proteins, have been found to possess tumor protease inhibiting properties. The natural protease inhibitors are not to be confused with the protease inhibitors used in antiretroviral therapy. Some viruses, with HIV/AIDS among them, depend on proteases in their reproductive cycle. Thus, protease inhibitors are developed as antiviral means.

https://en.wikipedia.org/wiki/Protease

ZIKA virus. Original, experimental method of therapy.

Venoms. Certain types of venom, such as those produced by

venomous snakes, can also cause proteolysis. These venoms are, in fact, complex digestive fluids that begin their work outside of the body. Proteolytic venoms cause a wide range of toxic effects, including effects that are:

cytotoxic (cell-destroying) hemotoxic (blood-destroying) myotoxic (muscle-destroying) hemorrhagic (bleeding)

https://en.wikipedia.org/wiki/Proteolysis#Venoms

ZIKA virus. Original, experimental method of therapy.

Inhibitors of Serine Protease. Inhibitors of Cysteine Protease. Inhibitors of Metallo-Proteases. Inhibitors of Aspartic Proteases.

ZIKA virus. Original, experimental method of therapy.

Example of Inhibitors of proteases. http://iti.stanford.edu/content/dam/sm/iti/documents/himc

/immunoassays/ProteaseInhibitionGuide.pdf Classes of Protease Inhibitors available from Roche

Applied Science

ZIKA virus. Original, experimental method of therapy.

Protease Inhibitors can significantly reduce clinical symptoms of ZIKA virus disease and complications.

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

MAC - Membrane Attack Complex Inhibition with monoclonal antibodies.

The membrane attack complex (MAC) or terminal complement complex (TCC) is a structure typically formed on the surface ofpathogenic bacterial cells as a result of the activation of the host's alternative pathway, classical pathway, or lectin pathway of thecomplement system, and it is one of the effector proteins of the immune system. The membrane-attack complex (MAC) forms transmembrane channels. These channels disrupt the cell membrane of target cells, leading to cell lysis and death

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

MAC - Membrane Attack Complex Inhibition with monoclonal antibodies significantly reduce symptoms and biological sequelae of Zika virus Diseases.

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

A number of proteins participate in the assembly of the MAC. Freshly activated C5b binds to C6 to form a C5b-6 complex, then to C7 forming the C5b-6-7 complex. The C5b-6-7 complex binds to C8, which is composed of three chains (alpha, beta, and gamma), thus forming the C5b-6-7-8 complex. C5b-6-7-8 subsequently binds to C9 and acts as a catalyst in the polymerization of C9.

Active MAC has a subunit composition of C5b-C6-C7-C8-C9{n}.

https://en.wikipedia.org/wiki/Complement_membrane_attack_complex

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

It is composed of a complex of four complement proteins (C5b, C6, C7, and C8) that bind to the outer surface of the plasma membrane, and many copies of a fifth protein (C9) that hook up to one another, forming a ring in the membrane. C6-C9 all contain a common MACPFdomain. This region is homologous to cholesterol-dependent cytolysins from Gram-positive bacteria.

The ring structure formed by C9 is a pore in the membrane that allows free diffusion of molecules in and out of the cell. If enough pores form, the cell is no longer able to survive.

If the pre-MAC complexes of C5b-7, C5b-8 or C5b-9 do not insert into a membrane, they can form inactive complexes with Protein S (sC5b-7, sC5b-8 and sC5b-9). These fluid phase complexes do not bind to cell membranes and are ultimately scavenged by clusterin and vitronectin, two regulators of complement

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

CD59 acts to inhibit the complex. This exists on body cells to protect them from MAC. A rare condition, paroxysmal nocturnal haemoglobinuria, results in red cells that lack CD59. These red cells can, therefore, be lysed by MAC.

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

The complement system functions as an immune surveillance system that rapidly responds to infection. Activation of the complement system by specific recognition pathways triggers a protease cascade, generating cleavage products that function to eliminate pathogens, regulate inflammatory responses, and shape adaptive immune responses. However, when dysregulated, these powerful functions can become destructive and the complement system has been implicated as a pathogenic effector in numerous diseases, including infectious diseases. This review highlights recent discoveries that have identified critical roles for the complement system in the pathogenesis of viral infection.

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

Complement and viral pathogenesis Virology Volume 411, Issue 2, 15 March 2011, Pages 362–373 Special Reviews Issue 2011 Kristina A. Stoermer Thomas E. Morrisona

ZIKA virus. ZIKA virus. Original, experimental method of therapy.

CD59 is a cell surface protein that prevents formation of the terminal membrane attack complex (MAC) involved in membrane disruption and cell lysis.