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Cytokines in the immune system Audai Ma'aiah Ziad Al-Nasser Monday, 1/8/2011

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Immunity: lecture 21 Date: Monday 1-8-2011 Done by : audai ma'aiah

Chapter :23 Yesterday if you remember, we talked about apoptosis and anti apoptotic pathways , and even what happens if you have a defect in the genes that control the anti apoptotic pathways , I gave you an example of an autoimmune phenomena , we call the it the autoimmune lymph proliferative syndrome , where if you have a defect in the anti proliferative and anti apoptotic pathway, how the pathway is going on and no apoptotic pathways is taking place, so you will have a plenty of b-cells are produced and lymphoma will develop out of the b-cells. Then we started talking about the cytokines, and we said that cytokines are protein molecules that are produced from the innate and adaptive immune system, and their main goal is to communicate between the adaptive and innate immune systems. And we said that those proteins are produced from cells, and theoe cells are not necessarily immune cells, they can be produced from other types of cells like skin ( keratocyte )and many other tissues. And they can act on immune cells and other types of cells and thats why we call them cytokines. So they are molecules, and we must have receptors for them, but if we do not have the molecules or the receptors , the patient will be immunocompromized . and we said that the mechanism of immune response needs the presence of these cytokines to take place, and we said we can use these cytokines for treatment, also we can block the receptor or neutralize the cytokines in certain diseases . also we said that cytokines are produced in short term, when they are needed, but when they are not needed , then their reaction will stop. and this is really very important as we said with IL-2, and how IL-2 induces an immune response , stimulation of the cell and the cell is going to stop in response to that, other wise they are going to stay activated and we dont need that. We said also that the bulk of these cytokines are up-regulators (enhance) or down-regulators (suppress) , and we should know that all the cytokine that are up-regulators and those that are down-regulators .

You should know two terms about cytokines, one we call redundancy, that means you can have more than one cytokines that have the same action, like interferon (gamma) , IL-1 and ILPage 2 of 13

6 in acute phase cytokines . the second one is pleiotropism , which means many cytokines is working on many cell types at the same type. Also you should differentiate between autocrine and paracrine, where autocrine the cytokine is produced and the receptore will develop on that cell, then the cytokine will work on the same cell that has produce it. While the paracrine , the cytokine is produced by a cell, and the effect will be seen on a neighboring cell . these cytokine usually are produced in low amount, but when we have severe infection , we can have large amount of them. So we can measure the amount of cytokines as an indication for infectious process or inflammatory process, like granuloma. We talked about interferon gamma and mycobacterium tuberculosis when we measure the interferon gamma we can find that we have activation of latent phase and we said how the nomenclature and how we call them ,

for example , interferon they interfere with viral replication . interleukins they come out of white blood cells, and for the numbers they refer to the time of discover. Chemokines the ones that act as chemotactic factor . IL-10 is a very important down-regulator cytokine , it helps in the regulation of the immune response and iso-type switching to IgA antibodies. we have also (TGF-beta) it act as downregulator that can suppress the immune response. you should focus on the fact that we have to measure the increase in cytokine amount as an indication of inflammatory process such as (TNF, IL-6) in septic shock . Treatment : also we can use the cytokines for treatment , such as interferon (alpha) in the treatment of hepatitis B , and its side effect is so minimal , and its mechanism is interfering with replication of hepatitis B virus , while here when we have interferon (gamma) that is activated it leads to necrosis process and activation of macrophages and so on where damage could take place and liver failure could come out . Cytokine are (pleiotropic ) affecting many different types of cells and mild side effect is common , also we should know that adaptive immune system cytokines is secreted at low level and are effective over short ranges , and then they need to be disappeared for the process to be finished. High doses usually causes severe side effects and we can do blockage , we mean by that block them in auto immune diseases ,TNF for example that we use for block its receptor or for neutralizing .IL-1 and TNF ,they have almost the same mechanism of action in RH ,so we can block them .redundancy ,we have the same mechanism of action ,many cytokine have the same effect, so if we block one ,the other might still be working ,so some time these processes might not be effective as we said .
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We can interfere with IL-2 (Most important proliferative and differentiating cytokine to immune cells) signaling in transplantation to prevent rejection by: Ciclosporin, which bind with immunophillin to calcineurin and inhibit IL-2 signaling, and Basiliximab which bind and block the IL-2 receptor Cytokines Receptors: 1. 2. 3. work as BCR and TCR in a singling pathway they are 3 types General chemokine receptor family The chemokine receptor TNF receptor They have similar structure; so they called hemopoiten receptors Receptors are up regulated after cell stimulation. General chemokine receptor family: They share structure composed of Transmembrane molecules with extracellular domains IL-2 (Proliferative to other cells), IL-4 (induce TH2 formation), and IL-7 (T-Cell proliferation and differentiation) have Receptors that share with 3 polypeptide chains, include chain.. if chain is defective X-SCID disease. (X-linked sever combined immunodeficiency disease) Steps for general chemokine family receptors activation: Cytokine binding to its receptor Aggregation as in BCR and TCR JAKs (cytoplasmic parts) activation by phosporylation STATs Activation and dimerization by phophoryaltion Activation of transcription factors o Ex: IL-2 binding will lead to JAK-5 activation, that will lead to STAT-1, and STAT-3 activation

1. 2. 3. 4. 5.

The Dr read this Table

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Pro inflammatory cytokines: give signal that we have been attackedmainly by macrophages

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The Dr said that we have to remember that CTLs (in fig above) need 2 co stimulatory signals: one from Ag-MHC complex binding to CD-8, and the other from Cytokines (IL-2).then the Dr. repeat the mechanism of activation:This is what I was taking about regarding here the cytokines that they come from, for example ,from T helper cell .here are the cytokines they have to act on T cytotoxic cells to initiate the signal ,thats mean one signal initiated by the presentation of the antigen by class 1 MHC and the second signal it has to come from the T helper cell ,so how the T cytotoxic cells going to be activated to produce the preference ,cytolysin of the granazine that will activate here the apoptotic pathway . when the cytokine act on the cell ,the receptor start to develop and they act on the same cell that they develop the receptor .this is the autocrine affect. the JAKs here (triangles) they will come and bind through the phosphorylation mechanism and they are going to be activated following the aggregation of the IL-2 receptors (similar to the aggregation of the TCR or Igs).so the JAKs is going to be activated ,and the JAKs activation stimulate the polymerization of the STATs molecules as you can see ,and the activated STATs will stimulate the transcription factors .STAT dimer will migrate to the nucleus and initiate transcription of genes required for T cytotoxic cell proliferation and the production of the cytolysin and the granazine as I said .

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So the cytokines produced the act on areceptors ,the receptors will aggregate ,they stimulate the JAKS ,then JAKs will stimulate the STATs ,and the STATs will go to the nucleus and activate transcription factors and the process will go on .and as I said for the IL-2 ,its JAKs-5 ,STAT-1 AND STAT-3.this is the combination ,and each cytokines have adifferent JAKs and STATs that is going to be stimulated . If the IL-2 is not needed, then the signaling will stop, we don't need to continuously activate the receptor. What about the chemokines.the chemokines they have so many of those and there receptor ,we have around 50 chemokines and we have around 20 receptors ,so more than one type of the chemokines will bind to the same receptor ,they bind and here the mechanism of the chemokines are different,they dont have JAKs and STATs ,here just simply the GDP is going to be replaced to GTP(guanine triphosphate) ,and this will going to activate the cell to go into the area where its supposed to go .for example some of these chemokines receptors are coreceptor molecule that the HIV to bind ,one of them we call it the CHCR4 and CCR5 .those are the ones where HIV virus can bind in addition to the CD4 so we have two receptor for the HIV virus ,one is the CD4 and the other is the chemokine receptor CHCR4 AND CCR5 . The third group of the cytokines are the TNF and TNF receptor ,and those are involved in the apoptotic pathway and also they can be an up regulator as well at the same time .we have TNF can be secreted or it can be membrane bound ,and the one that is membrane bound will act in the apoptotic pathway ,it binds to other cell that has receptor, or the free one here it can act as an acute phase response .and we have also the CD40 and the CD40 ligand, and the combination between the CD40 and the CD40 ligand also can activate the cells. and we have talked about those if you remember at the APCs and the T helper cells and how those are so important, and if the CD154 (CD40 ligand) is missing ,the patient is going to have the hyper IgM syndrome .Fas and Fas ligand is also used, it depend on the target cell they bind to.so here we talked about three mechanisms ,TNF and its receptor,CD40 and its receptor as will as the Fas and Fas ligand receptor . Chemokines receptor are specialized molecule that cross the cell membrane 7 times ,so we will have part outside the cell and part inside the cell like those of Igs and TCRs in away.

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As you see that both the TNF and Fas are induce apoptosis, and the TNF and CD40 are induce gene transcription.

Again here we can see the mechanism of each one . So these similarities and differences I think you should know .
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Look here into the interactions of these cytokines and the bacteria ,for example here we have been invaded with bacteria in a tissue as you can see,so the tissue can produce cytokines or they can bind into macrophages (toll like receptor)so the toll like receptor they produce cytokines with the same type, these cytokines that is produced in the tissue or from the macrophages they can act as you can see on the APCs here as will as on the T helper cell at the same time ,so the T helper cell is going to be activated and produce cytokines that have so many functions ,for example some of them in the mucous membrane can induce the production of IgA antibodies like for example the TGF-beta the can induce isotype switching to IgA .and the same thing here if we required the production of the Ig so these cytokines (IL-12) will make the switch of TH-1 into TH-2 ,and TH-2 will act on mast cells (by IL-3, IL-4, IL-5) to produce IgE which bring eosinphils into the area,so it depends on the nature. and when TH-1 is activated ,it will suppress the TH-2 and vise versa . so we can see the role of the cytokines in the communication between the innate and the adaptive immune response and how they up regulate for a short period and then they will be suppressed ,if they are increased in Avery large amount ,they could act as hormones .

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Slide 13 + 14 :So the role of cytokines in the immune response here as I said start with the acute inflammation (danger signal) ,activation of the toll like receptor ,IL-1 ,IL-6 ,G-CSF,TNF and IL-8 those are the initial cytokines that are produced mainly from macrophages that we have been attacked ,"acute phase response". In viral infection ,interferons and T-helper 1 activation will occur ,then we end in a state we call it T cell priming (when the cell gets exposed to the cytokines ,it gets priming).now the APCs cause stimulatory molecules CD40,CD80,ICAM (immunological synapse) and type 1 IFN leads to IL-2 production (autocrine and paracrine) ,and then the cells are going to be activated . Development of specialized T-cell response at the GIT ,the TGF-beta switch to IgA antibodies then inhibition(down regulator).TH-1 response ,APCs ,IL-12 ,type 1 IFN ,IFN-gamma , IgG switch because we need that for opsonization as we said .IL-6 ,stimulatory Igs stimulate B cells,TNF also stimulatory macrophages and the development of granuloma . TH-2 respons if we are invaded by parasite ,Tcell transcription factor,so befor we get the IL-4 , we get a protein we call in GATA-3 and for the TGF-beta we call it T-bet ,so this is like an intermediate stage in the development or the activation of those cells ,GATA-3 here for the development of TH-2 and the production of IL-4 and IgE antibodies and mast cells,so GATA-3 has to do with mast cells and IgE antibodies and so on . IL-3 ,IL-5, and the eotaxin ,and im sure you know this type of relation ship ,here we just added an intermediate step we call it GATA-3 and T-helper 1 we call it the T-bet for the development of the immunosuppressive cytokines . The end of the immune response ,the danger signal will fall down ,less APCs presentation ,decline T cell stimulation, low IL-2, low Bcl-2 expression ,and more apoptosis pathway. Remember the inhibitory cytokines IL-10 and the TGF-beta ,and those there role in tolerance we talked about that before ,and the memory maintenance IL-7 ,its production in the bone marrow and its significant in memory and the nurturing of so many other cells at the same time ,and the neutrophils number by the G-CSF . CHAPTER :24 So now you can see how these cytokines and you should know by now all those names ,when I say IL-1 ,IL-2 ,IL-3,IL-4,IL-5,IL-6,IL-7,IL-8, we haven't talked about IL-9, we talked about IL-10, we haven't talked about IL-11, we talked about IL-12 , and we have IL-18 which is very similar to ILPage 10 of 13

12 .you should remember all of them ,by names ,where they come from, each one ,its function , where they going to act . interferons alpha ,beta ,and gamma ,very important to remember ,and we talked about the TGF-beta as suppressor or down regulator cytokine. We taked about TNF and its dual effect on the apoptotic pathway as an acute phase response in the granuloma and activation of macrophages (very important to remember that),and we talked about the Fas Fas ligand in the apoptotic pathway .the idea is very simple, these are the terminology that we are using to describe as you can see the immune physiological process ,up regulation ,down regulation .and I want you to know when you hear about these terms here in any immunology book, you can understand what you are talking about . we will keep adding up many of these terminology here in this course, we still have more to come ,but things that will be coming up now its like an application of most of these basic information that we have been taking about ,like for example today we will be talking about infections and vaccines and you will see how these simple applications of what we have been talking about ,its just we will add few information ,maybe little bit that you don't know, but the plug of what we are going to talk about you know. And you will see how you are going to use these terminology that you are learned over the passed weeks in understanding these applications. For example here ,we will be talking about infections and immunity ,and the role of vaccine and vaccination related to. we have an adaptive immune response ,and because we have pathogens that are involve ,we can stimulate this natural infection or we can induce the primary immune response . so when we get infected in the first time, then the cell become primed, and we can primed the lymphoid cells by exposing them to antigens of pathogens ,as simple as that, so we can make vaccines. Pathogens when they invade ,successful pathogens they have as I told you before and we keep repeating that ,they have evasion mechanism to stay in our body for a longer period of time ,and to reflect more injury ,and our body is going to counter attract again measures to get rid of those ,so in infection sometimes we could have more than one immune mechanism and we talked about that if you remember capsulated bacteria when it enters our bodies ,the capsule interfere with phagocitosis ,this is a defense that the bacteria is using as evasion mechanism ,our body is going to respond back by the development of antibodies against the capsule ,and the antibodies will bind to the capsule and then the bacteria is going to be killed by complement or by phagositosis ,opsonization and so on.

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And these are the different immune mechanism that we keep talking about here as time will come. The vaccines to by pass the primary immune response and to prime those cells ,and to have memory cells in general, the best vaccine that we are going to use are what we call the live vaccine ,we use what we call live attenuated vaccine . we want to stimulate the natural infection as much as we can ,but some times these live attenuated vaccines they could revert back to virulence ,the could revert back to the wild strain (use the term wild strains and mutant strains) ,wild strains are the one that causes the disease ,the mutant is the attenuated one ,so these attenuated ones they could revert back to virulence and cause the disease again ,so you have to way the risk here and but contraindications which people they should not taking the live vaccine. Other vaccines here are the killed vaccines as long as antigens are there ,we killed it and we give it to people ,these are safe but not as effective as the live vaccine . or sometimes we say why to expose people to the whole bacterial antigen (we could taking the attachment proteins alone and we can give it to people ),so we can use what we call subunit vaccine or we can take for example the capsule of the streptococcus pneumonia or noplus (not sure) influenza type B .why are live attenuated vaccine are the best? 1st: they stimulate the natural infection ,so if for example polio militis I get infected through my oral cavity , then I can take the live attenuated polio vaccine in my oral cavity to the GIT and makes local immunity in the GIT . 2nd: live attenuated vaccine they replicate in our bodies, so they give us Avery large dose of the antigen ,so the antigen is present in our bodies for a longer period of time .so our immune system (B and T cells ) are primed continuously . 3rd: those organisms ,they will be shit if they infect our GIT ,they will be shit like our GIT outside, and they will infect other people ,and this is if you remember the heard immunity ( ),every body will be vaccinated naturally . the only side effect for that is reverse virulence and they revert back to their virulent factor (to the wild strains).so the mechanisms of immunity here "infections and vaccination", the goal is to achieve sterilizing immunity ,and we are not successful all the time to achieve sterilizing immunity all the time, some times we have breaches in our immune system . we start from mother to child Igs ,they comes from the mother IgG antibodies to the baby, we call that natural passive immunity . the 1st collastrim IgA antibodies they come from the mother to the child ,this is the first thing that we have .the 1st exposure the baby Is going to have here is the primary immune response
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and memory cells will develop.we will try our best to stick that base by not exposing the baby with the wild strain ,the attenuating strain (the antigens of the baby). small pox ,poilo milltis ,diphtheria and herpeses are the most effective and successful vaccines in maintain memory. those vaccines have safe the life's of millions of people all over the world, and sure you have hear about small pox where the last case we saw the small pox was in 1977, and they had an accident in 1981.but after that we never hered about small pox .why small pox managed to be eradicated completely ? 1st: small pox infect humans only, doesn't infect animals, so if you vaccinate humans, that set you will not get it again. Its from human to human, no animals are involved. 2nd: the virus antigenic genetically stable ,it dosen't mutate from one strain to another so we don't expect to see many strains that they will come from other viruses that are similar . 3rd: doesn't transfer by insects or snacks or other animals. What about poilo militis ,the same thing almost we have 3 strains ,poilo 1 , 2 and 3.and almost we are getting so closed here to the eradication of poilo militis. Diphtheria ,the last case was in 1993 .whooping cough ,the same thing here we haven't hear about those after the 1980s. Finally ,special thanks to Dr.baha2 al 3zam and Dr ghassan shbool for their help .

DONE BY : Audai Ma'aiah

THE END
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