Bag. Biomedik/ Bag. Patologi Anatomi Fakultas Kedokteran Universitas Sebelas Maret Antigen recognition Presentasi antigen by MHC class II or class I T receptor & accessory molecules
EXPERIMENTAL MODELS USED TO STUDY T- CELL ACTIVATION Parameter of T cell activated :
(a) early signal transduction events, such as protein tyrosine phosphorylation or an increase in cytoplasmic free calcium ([Ca2+]i), that do not necessarily lead to a cellular response; (b) expression of new cell surface activation antigens, Including the a chain (CD25) of the IL-2 receptor (IL- 2R), the transferrin receptor, class II MHC molecules on human T cells, and CD69, a molecule with as yet unknown function; (c) production of lymphokines, such as IL-2 or IL-4; (d) cell proliferation; and (e) cytolytic activity. Requirements for the Initiation of T- Cell Activation Aktivasi sel T diinisiasi oleh adanya : interaksi antar sel . Spesifik antigen dengan TCR mengawali interaksi antara sel T dan APC. Antigenic peptides bound to MHC molecules on APCs are recognized by T cells bearing antigen-specific TCRs. Disamping itu ikatan antara ke dua sel ini juga diperkuat dengan adanya ikatan beberapa molekul lain (gambar 1). The TCR and other cell surface molecules contribute to the initiation of T- cell activation by inducing signal transduction events and by contributing to the overall avidity of the T cellAPC interaction. Considerable evidence has accumulated to suggest that at least one molecule, a co-stimulatory receptor, must initiate signal transduction events distinct from the TCR in order to initiate IL-2 secretion and induce a proliferative response in naive T cells.
T CELL APC or TARGET CELL T CELL TERAKTIVASI SEKRESI IL-2 ( T CELL Growth factor) EKSPRESI IL-2 RECEPTOR PROLIFERASI & DIFERENSIASI SEL T Event Example Cellcell interaction T cellAPC CTLTarget Cell Receptor-ligand binding TCR-Antigen/MHC Transmembrane signal transduction Activation of Lck Generation of second messengers 1,4,5-IP3 and DG Second messenger effects Ca2+ mobilization Protein kinase C activation Biochemical pathways Phosphatidylinositol pathway Ras pathway Cellular events MTOC reorganization Secretion of cytolytic granules Early gene activation c-myc, c-fos Intermediate gene activation Lymphokines, lymphokine receptors, nutrient receptors Late gene activation Genes involved in cell proliferation, 4F2,VLA-2 MAJOR EVENTS INVOLVED IN T CELL ACTIVATION T CELL ANTIGEN RECEPTOR Figure 3. Interaksi antara sel T dan APC yang berujung pada produksi IL-2. Interaksi seguensial antara TCR dengan peptida antigen-MHC Complex, memacu ekspresi protein CD40L pada sel T yang berinteraksi dengan CD40 pada permukaan APC. Hal ini menginduksi ekspresi molekul B7 pada APC yang dapat menstimulasi CD28 yg mrpkn co-stimulatory receptor pd sel T. Kedua signal ini bersama-sanma menginduksi ekspresi gen IL-2. Stimulasi produksi IL-2 The role of IL-2 in REGULATION OF T- CELL PROLIFERATION Antigen-activated T cells that produce IL-2 can : (a) promote their own clonal expansion, (b) promote the proliferation of other T cells that are activated by the same or a related specific antigen but can not produce IL-2 (i.e., CD8+ cells), (c) promote the expansion of previously stimulated cells that express low levels of high-affinity IL-2Rs (i.e., memory T cells), and (d) promote the proliferation of non-T cells that express IL-2Rs (i.e., B cells or NK cells). In addition to its growth-promoting effects on various cell populations, IL-2 can influence the development of various differentiated functional activities. Stimulation of the TCR induces naive T cells to progress from the G0 to the G1 stage of the cell cycle and to express high- affinity IL-2s The function of IL-2 is to promote further progression through the cell cycle. The ability of IL-2 to induce cell cycle progression, from G1 through S, G2, and M, depends on the binding of IL-2 to its high-affinity receptor. Although IL-2 may not be the only lymphokine that can induce T-cell proliferation, the importance of IL-2 in promoting such cell cycle progression in peripheral T cells is underscored by the decreased proliferative response to mitogenic and antigen stimulation of T cells from mice in which the IL-2 has been disrupted by homologous recombination. Hence, the binding of IL-2 to its receptor and the ensuing signal transduction events play central roles in most immune responses. The role of IL-2 in REGULATION OF T- CELL PROLIFERATION IL-2 Reseptor Setidaknya ada 3 jenis rantai IL-2 reseptor
1. Rantai CD25, the IL-2R chain, is a 55-kD integral membrane glycoprotein that has an affinity constant (KD) of 10-8 M. This low-affinity form of the IL-2R comprises the most abundant form of the IL-2R expressed on activated T cells and human T-cell leukemia virustransformed lines.
2. Rantai : a 70-kD glycoprotein, binds IL-2 with intermediate affinity (KD of approximately 10-9 M) when it is expressed on T cells.
3. Rantai : The IL-2R chain is a 64-kD integral membrane protein.
The and chains of the IL-2R are members of a family of cytokine receptors that have related structural features in their extracellular domains The high-affinity IL-2binding site appears to result from the combined characteristics of the , b, and g chains.
Model Reseptor IL-2
SIGNAL TRANSDUCTION BY THE T-CELL ANTIGEN RECEPTOR Src PTKs Involved in TCR Signal Transduction
Lck and Fyn are the major Src family PTKs expressed in T cells. Both have been implicated in interactions with ITAMs and in TCR signal transduction. Prior to reviewing the specific role of each of these kinases, the overall common structural features (Fig. 5). The Src kinases vary from approximately 50 to 60 kDa172. At the N-terminus of each of these kinases, at position 2, is a glycine residue that is myristoylated. This allows for membrane attachment. Some of the Src kinases, including Lck, are also palmitoylated at one or two cysteine residues contained within the first ten residues, and this modification may be dynamically regulated. This further facilitates membrane localization, particularly the plasma membrane. Within the N-terminal, 40 to 70 residues are also the most distinguishing sequences among this family that probably play important roles in the unique functions and interactions of each of these kinases. The unique region is followed by the Src homology 3 (SH3) domain, which consists of approximately 60 residues. The SH3 domain is involved in directing proteinprotein interactions by binding in a sequence specific context to residues contained in proline-rich regions. The SH3 domain is followed by a 100-amino acid structural domain, the SH2 domain. The SH2 domain also is involved in proteinprotein interactions by binding to phosphorylated tyrosine residues contained in a particular sequence- specific context. Inhibitors used to study T-cell activation Inhibitor Target Neomycin Phosphatidylinositol turnover Lithium Inositol phosphate phosphatase H7 Protein kinase C Sphingosine Protein kinase C Staurosporine Protein kinase C Genestein Tyrosine kinase Herbimycin A Tyrosine kinase Tyrphostin Tyrosine kinase Vanadate Tyrosine phosphatase Phenylarsine oxide Tyrosine phosphatase (?) EDTA Ca2+ and Mg2+ EGTA Ca2+ Dimethylamiloride Na+/H+ antiporter Glucocorticoids Glucocorticoid receptor; diverse effects Cyclosporin A Calcineurin FK506 Calcineurin Rapamycin mTor Wortmannin Phosphatidylinositol 3-kinase LY294002 Phosphatidylinositol 3-kinase THE REGULATION OF T-CELL PROLIFERATION Other Mechanisms Regulating T-Cell Growth
IL-4 and IL-15 are the most likely to function as T-cell growth factors in the absence of IL-2.