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Cell-Cell Interaction in Generating Effector Lymphocytes

Tamara Shoter Ziad Al-Nasser Thursday, 21/7/2011

Immunology Lecture #14 Thursday, 21/7/2011 Done by: Tamara Shoter The doctor started the lecture talking about the exam; its place, time...Etc. So, yesterday we were talking about the process of T-cell development and we said that it occurs within the thymus gland. The earliest thymocytes (Double negative cells, which express self-surface molecules such as CD25 and CD44) enter the subcapsular zone, where they start to mature. After that, they move further into the cortex where we have MHC antigens. Now, in the cortex, double negative cells mature into double positive cells (CD4+ CD8+). Thymocytes' fate depends on the ability of their receptors to recognize class I or class II MHC molecules. Double positive (DP) cells that recognize calss I MHC molecules will become CD4 + cells, while DP that recognize class II MHC molecules will become CD8+ cells (this process is called Positive Selection and it occurs in the medulla).

Notes:
1. T-cells that have high affinity for self-antigens are going to be eliminated. 2. There's a mechanism called central tolerance related to the endocrine gland, where all the tissues in our bodies supposed to be presented in the thymus gland but it doesn't happen this way, so we have (?). The auto regulatory immune gene can control this process with selfrestriction and if we have a mutation in this gene, then self tolerance or self restriction is going to be affected and we call this autoimmune polyendocrine syndrome I. ( Multiple endocrine organs that have an autoimmune disease). 3. Some self-reactive cells may escape into the periphery and this will cause the development of something called 'Peripheral Tolerance'.

Also, if you don't have accessory molecules like CD28 which develops at the periphery, then peripheral tolerance will develop (Not fully sure). 4. We talked about T reg (known as suppressor T cells) which produce suppressive cytokines like; IL-10 and the transforming growth factor beta (TGF-). As I said the gene that codes for that is called Foxp3. Mutations in it will result in a multisystem autoimmune disease. 5. 95% DP cells never mature because they lack TCRs that can appropriately recognize self-MHC.

T helper cells are divided to Th1 and Th2 cells based on the nature of the infectious process or the mechanism of the immune destruction that we need. For example, if we need to destroy virally infected cells or tumor cells, we require Interferon-gamma (IFN-) which activates macrophages that express class I MHC that is need for cytotoxic T cells. Remember, tumour necrosis factor and interleukin 6 cause elevated body temperature and isotype switching to (IgG) which acts as an opsonin. Macrophages produce interleukin 12 which stimulates the

differentiation of Th0 to Th1 cells. The same thing here, we need other types of antibodies we call them IgH (IgH means any antibody that's not IgM, be it IgG, IgE ..Etc). Now, Interleukin 4 is produced by mast cells which induce differentiation of Th0 cells to Th2 cells, which will also produce interleukin 4 and interleukin 5, and interleukin 5 attracts eosinophils. We also talked about the gamma delta T-cell subset which represents a small percentage (5%) of all T cells. The gamma delta T-cell receptor can recognize certain peptide and non-peptide antigens without processing and in the absence of MHC class I or class II molecules. These cells help other T cells to activate macrophages, also they are involved in the lysis of virally infected cells and they are a part of the first line of defence. (Skin, gut mucosa..) So here, in DiGeorge syndrome, the 3rd and 4th pharyngeal pouches aren't developed. Therefore, the thymus and parathyroid glands will be missing, of course patients with this syndrome will have a very low number of T cells. However, the B cell count is normal .Since these patients have T lymphocyte deficiency, they will be more susceptible to viruses and tumours. There's a virus called Epstein-Barr (EB) virus that binds to CD21 receptors present on the surface of B cells and this will cause differentiation of B cells into antibody-producing plasma cells, which in turn activate the CD8+ cells that are going to destroy infected cells. Also, TNF and Il-6 cause elevated body temperature. See the figure below:

Chapter 16 - Cell-Cell Interaction in Generating Effector Lymphocytes

So now the B, T cells left the thymus gland and the bone marrow as nave/ virgin lymphocytes and we need those to be primed in order to stay in the secondary lymphoid organs as long as we want an efficient immune system. If you are following the development of the B cells & T cells and the markers that will develop on their surface when they go to the secondary lymphoid organs, we see that after they go to the secondary lymphoid organs and interact with an antigen we see other markers that are so important for the activation of those B or T cells. We can see that the binding of the antigen -when it's carried by the MHC antigens- to the receptor is providing me with one signal but you need other signals, and those come from the accessory molecules and those are so important; sometimes for cytokine function, for isotype switching. So the immune system will not be completed unless those accessory molecules that I am going to talk about are there and perform their function, if they are missing then we are going to be immunocompromised .This is what I am going to talk about in Cell-Cell Interaction In Generating Effector Lymphocytes. We are talking about an antigen taken by an APC, processed, presented to the surface and then binding to the B & T cells; this is one factor, we have other factors/ signals that are going to be involved in the secondary lymphoid organs. Refer to the Over-view figure at the beginning of the chapter: We can see here in the Fig. the antigen taken by an APC (through the T helper cells), T helper cells will be activated & produce cytokines which act on the B cell, B cell will be differentiated into a plasma cell that will give AB. This process is called effector B-cell generation. We are talking

about thymus dependent; this is the role of thymus (the process of effector B cell generation in the involvement of T cell). This is the activation of B cells. So it's not enough to bind the Ag to a B cell, you require also the help of the thymus gland through CD4 by the production of cytokines to the B and to the differentiation to a plasma cell. The same thing here applies for T cytotoxic and the killing, it's not just enough that the virus or the tumor Ag is presented to the T cytotoxic cell through class-1 MHC Ag; this is one signal, the 2nd signal must come also from a T helper cell. We will see the cytokines that are going to be produced; sometimes if the Ag presenting cell is already infected, certain viruses, extracellular vaccines could be presented by class-1 MHC Ag to T helper. So both (?) can provide cytokines through this process that will activate the CD8 to change into an effector cell and then produce the cytolysin & kill the target cell. (You better skip this paragraph). So as you can see here (in the fig), in both* the T helper are required for AB production or for killing process. * Effector B cell generation & effector T cell generation. T cell recognizes antigen that is only presented by an MHC Ag on the APC (thymus dependent Ag). While B cell doesn't have to. Co-stimulatory signals provided by the interaction with accessory cells, so we require other signals beside the binding itself. The outcome is plasma cells -as effector- that give me antibody OR effector T cell either helper or cytotoxic. The helper will be activated and act on the T cytotoxic activating it when this cytotoxic is bound to the Ag of the surface of the cell that I need to kill.

Refer to Fig.16.1/Pg.121: So this is what's happening; here a B cell, it has a B cell receptor and the Ag is bound here (to the BCR) will be taken inside, bind to class-2 MHC Ag and then be presented (so B cell acts as an APC) to the T-helper. Then the T-helper is supposed to be activated. Now -as I said- when this is taking place we require other mechanisms for the production of cytokines. Now this stage of T cell activation, makes the T cell (the prime T we call it) form other accessory molecules that are needed for the production of cytokines and for the activation of the T helper cell that's going to work on the B cell, that's going to differentiate into plasma cell, that give antibodies. Fig. 16.2/Pg.121: Look for example: This is Lymphocyte Functional Antigen on T cell that's going to bind to the ICAM-1 on APC. CD28 & CD80(B7); this interaction is so important to tell you that this stage is a thymus dependent. CD154 (CD40L) is going to bind to CD40 on the APC. This binding makes the cytokines that are needed for isotype switching; so if this binding is not formed only the B cell will produce the IgM (as if it's thymus independent). * We have a syndrome regarding this called X-linked hyper IgM syndrome; the patient keeps producing IgM and doesn't produce IgG and you know how important the IgG is for opsonization, neutralization and the high affinity of Igs and so on -while the IgM has a low binding affinity. So this syndrome results from the missing of CD154, and how can we treat those? By Gene Therapy; we have to introduce the gene that's missing so the isotype switching is going to take place.

*All these interactions occur when you have an APC in the paracortical area of the lymph nodes. *If any of the up-mentioned receptors is missing then we become immune compromised. Fig. 16.7/ Pg.125: These are the differences between TH1 and TH2, we talked about that: TH1: interferon- and the tumor necrosis factorresponsible for fever, & IL-6 as well as IL-10. IL-4 & IL-5 not produced here. TH2: IL-4 & IL-5 and no INF- & TNF- .

Fig. 16.4/ Pg.122: Look here into the complete picture that I was telling you about; class-2 presenting the Ag to the T helper providing the 1st signal. CD3 is so important in the T helper for the passage of the signal. And then the CD28 & the CD80 (B7) the same thing; give me an indication that this is thymus dependent. And we talked about the CD40L and the CD40 that will give a signal here for the isotype switching as well. For B cell activation, it's not enough just to bind the Ag to BCR, this gives you just one signal & you require other signal as we said many times. This all doesn't happen in the bone marrow it happens when the cell is primed; when the cells go to the secondary lymphoid organs & meet the APCs in the interaction. This is naturally what should happen. The explanation of stimulated or primed B & T cells in the secondary lymphoid organs is that the APCs are present in the secondary lymphoid organs so now what comes from the bone marrow or the thymus gland (Nave or virgin lymphocytes) are going to be stimulated there. The DC could produce cytokines, for example IL-12 for the differentiation of TH1 &TH2, those they make the HEV to produce the

selectins & the addressins & those are important for the stoppage of these lymphocytes & the passage through the assigned areas in the secondary lymphoid organs; the paracortical region or the follicular region. The adhesion molecules communicate through the cytokines, so the cytokines are also important for the adhesion molecules to develop. * The function of T helper cells is cytokines production; interleukins & other cytokines. * T helper 2 activation usually leads to antibody production of the IgE type and others (IgH: IgA, IgD, IgE), while IgG comes from T helper 1 and functions in opsonization. * T helper 1: cell mediated immunity, interferon gamma, isotype switching to IgG, then IL-6 and tumor necrosis factor. B cell can present Ag to TH cell by class-2 MHC Ag. Those you see between the APC at one side & the B or T cell on the other side we call them the immunological synapse. So when we talk about the immunological synapse, we are talking about the Ag presented with class-2, ICAM-1, CD40, CD80 (B7) & on the other side we have the receptor & the CD4, LFA-1, CD28, CD40L This relationship in the synapse is VERY important in isotype switching. So, what about Cd40 and Cd28? We said before that presentation with class II MHC antigen to the receptor will provide just one signal, the other signal should come from the accessory molecules in the synapse as we said. These

signals will stimulate T helper cells to produce

interleukin 2 which induces the differentiation of B cells. Also, interleukin 2 acts on a receptor called interleukin 2 receptor (Will be explained in details later on). Remember that interleukin 2 is the major cytokine needed for growth and differentiation, if it's not developed, we will be severely immunocompromised. And in order for interleukin 2 to be produced we need all the accessory molecules, so one signal here isn't enough. The co-stimulatory receptor-ligand pairs CD40-Cd154

(CD40L) and CD80 (B7)-CD28 act synergistically to enable T-H cell differentiation. Hyper-IgM syndrome results from mutations in the CD154 gene, here there will be no isotype switching to igG. Here, the doctor is talking about figure 16.6 on p.124, just follow the arrows. He added that cytokines are important for the development of memory cells.

Clinical Applications:
There's a disease called Leprosy ( ,) it's caused by Mycobacterium leprae. This disease affects the terminal (?), so you are going to have nodules and paraesthesia. They have noticed that if you have (?) infection, then lepromatous leprosy results, and if you have a localized infection we call that tuberculoid leprosy. In lepromatous leprosy, Th2 cells are going to be activated (Note: Mycobacterium tuberculosis activates Th1 cells which will activate macrophages..), antibodies will be produced (IgE type) which have no effect, so patients usually die. Now, in tuberculoid leprosy, small amounts of Th1 cells will be activated and interferon gamma, interleukin 6, and TNF will be produced, these cytokines will activate macrophagesEtc. (Note: Patients with tuberculoid leprosy have a positive skin test reaction) Also, patients can fully recover from this disease and they become immune to this bacterium. Refer to page p.126, Acid-fast bacilli are present in all parts of the body in lepromatous leprosy.

Refer to p.127, here is a patient with hyper IgM syndrome, he's immunocompromised because IgM antibodies have low affinity and no memory. So patients with this syndrome require gene therapy. Remember, the molecule that is missing here is CD154 or CD40L (L stands for ligand).

You can see how the cooperation is going to take place for the killing process as well. Now, activation of CD8 T cells in order to develop effector functions requires: (1) First signal: An antigen presented by class II MCH molecules. (2) Second signal: Interleukin 2 which we can get from the same virus that has been trafficked through the extracellular pathway or from T helper cells. (Note: We always require T helper cells in the thymus-dependent mechanism, to provide us with interleukin 2).

Notes: 1. In multiple scelrosis, sheaths around the axons will be destroyed by

Th1 cells. We notice that during pregnancy Th2 are going to be activated, so more antibodies will go to the baby. (Pregnancy reduces the activity of this disease Beneficial effect). Th1 and Th2 can't be activated at the same time; if we activate Th1 cells, we suppress Th2 cells and vice versa, we can use that in the treatment of autoimmune diseases.

2. We talked about conjugate vaccines against Streptococcus pneumoniae and Haemophilus influenzae B. The polysaccharides of these encapsulated bacteria stimulate thymus-independent antigens, resulting in the production of IgM antibodies only , so no memory cells will develop and there will be no isotype switching. So, how are going to deceive our immune system? We get the polysaccharide antigen and mix it with a protein. The protein component of the conjugate can then be processed and presented on the surface of B cell associated with MHC class II molecules; this complex is going to be recognized by T helper cells which in turn will produce cytokines needed for the production of memory cells.

Chapter 17 Immunological Memory

After Initial exposure to a certain antigen, memory cells will be formed and they will stay in our bodies for a long period of time. On a subsequent exposure (Secondry exposure) to the same antigen, there will be a more rapid and higher affinity immunological response and these memory cells will be released. Refer to figures (17.1, 17.2) They are very important. You have to remember that vaccines are used to create memory cells for pathogens before we encounter them. Also, there are qualitative and quantitative differences between naive cells and memory cells (Will be explained later on). Keep in mind that a single vaccine results in production of small amounts of memory cells, that's why it's important to give booster shots from time to time in order to achieve full immunity. (Since, the length of survival differs from one memory cell to another). One significant characteristic of memory cells is that they have anti-death genes antibodies which will neutralize the effect of death genes, so they can survive for a long period of time.

Note:
The expression of L-selectin (CD62L), the receptor that facilitates homing to peripheral lymph nodes in T memory cells is variable, because they are already activated, so they do need to go lymphoid tissues. Effector T cells low; because they aren't needed anymore, they are ready for action. Navie T cells High; because we need them to bind and go to the assigned areas.

Done!

I'd like to give a shout-out to my dearest friend; Mai Mazen. Thanks a bunch for helping me with this lecture :] Tamara Shoter