SPECIFIC IMMUNITY = ACQUIRED IMMUNITY = ADAPTIVE IMMUNITY SPECIFIC IMMUNITY - BASIC CHARACTERISTIC

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1 SPECIFIC IMMUNITY - BASIC CHARACTERISTIC SPECIFIC IMMUNITY = ACQUIRED IMMUNITY = ADAPTIVE IMMUNITY BASIC TERMINOLOGY SPECIFIC IMMUNITY humoral mediated with antibodies cellular mediated with T lymphocytes SPECIFIC IMMUNITY systemic in whole body local, mucosal on mucous membranes SPECIFIC IMMUNITY active induced during immune response (to the infection or vaccination) passive transfered with antibodies or cells ORIGIN OF SPECIFIC IMMUNITY Immune system of vertebrates is characterized by: Organization of lymphoid tissue of mezenchymal origin Existence of immunoglobulin superfamilly molecules Mechanisms of recombination of genes for diverzity Era of specific immunity began before 500 billion years genes for recombinases (RAG) were transfered horizontally from bacteria to the first chordates These recombinases start rearrangement of genes for variability

2 RECORDS IN IMMUNOLOGY Adult man produces about 5 billions of blood cells in his bone marrow dailly Lymphocyte (and sperm cell too) is the most rapidly dividing cell in the body Capacity of recognition of specific receptors for antigen is higher than the number of existing molecules in the nature THE MAIN FEATURES OF THE IMMUNE SYSTEM OF VERTEBRATES RECOGNITION OF SELF AND NON SELF SPECIFITY AND GREAT DIVERSITY OF RECOGNITION OF INDIVIDUL MOLECULES - ANTIGENS IMMUNOLOGICAL MEMORY THE MAIN COMPONENTS OF SPECIFIC IMMUNITY LYMPHOCYTE lymphocyte has the ability to compete with microorganisms short generation time high variability of antigen receptors T lymphocytes with T cell receptors B lymphocytes with B cell receptors ANTIBODY - IMMUNOGLOBULIN molecule composed with 4 chains (two heavy and two light) specificity is done by the hypervariability region CDR binding site for antigen one immunoglobulin molecule has two binding sites antibody with the same specificity exists in various izotypes - IgM, IgG, IgA, IgE Lymphocyte small cells with nucleus filling almost whole cell morfology small/large lymphocytes, lymphoblasts, plasmatic cells lymphocytes are involved in different processes of specific immunity can be divided into different subsets specifity of antigen recognising is mediated by receptors for antigen TCR and BCR

3 LYMPHOCYTES B lymphocytes differentiate into plasma-cells which produce antibodies T lymphocytes secure various function Th CD4+ helper secure regulation of immunity Tc CD8+ cytotoxic liquidate afected cells (with viruses, tumors etc.) double pozitive CD4+ CD8+ - helper memory cells Treg CD4+ CD25+ - inhibiting cells - supressors γδ T lymphocytes various function on mucous membranes FENOTYPIC MARKERS OF LYMPHOCYTES T LYMPHOCYTE CD3, (CD2, CD5), TCR to this main characteristics various subsets have their own molecules: TCRαβ lymphocytes: helper cytotoxic double positive regulatory TCRγδ lymphocytes : gamma/delta B LYMPHOCYTE CD4 CD8 CD4, CD8 CD4, CD25, FoxP3 ± expresion of CD8 etc. BCR, CD19, CD20, CD21, CD79, MHC II FLOW CYTOMETRY (SUBSETS IN DOGS) Th CD3 + CD5 + CD56 + APC SPECIFIC RECEPTORS FOR ANTIGEN CD4 + CD8 CD8 + CD21 + CD79α + γδ CD4 + CD8 +

4 SCHEMA OF SPECIFIC ANTIGEN RECOGNITION SPECIFIC RECEPTORS FOR ANTIGEN ON LYMPHOCYTES Igβ Igα BCR + CD79 Igα Igβ ζ ζ α β TCR + CD3 γ ε δ ε sigm TCR

5 ORIGIN OF BINDING SITE VARIABILITY Genetic background is created by - genes VDJ for variable part of heavy chain - genes VJ for variable part of light chain GENETIC BASE FOR BINDING SITE HIGH VARIABILITY primary antibody variability is created by combinatory joining of VDJ subgenes combination of heavy and light chains mutation inside subgenes allelic exclusion secondary antibody variability is created after contact with antigen genosomatic hypermutation During combinatory joining of subgenes V(D)J, gen C for constant part (determining isotypic characteristic) is attached GENETIC BACKGROUND OF HEAVY CHAIN ORIGIN OF BINDING SITE FOR ANTIGEN VH (~100) D1 - D30 JH1 - JH6 Cµ Cγ Cε Cα V5 D3 J2 Cµ Cγ Cε Cα germinal line B-Ly (IgM) RAG genes code recombinases for rearrangement Rearrangement of genes VDJ (VJ) code hypervariability region TCR on T lymphocyte BCR on B lymphocyte Binding site on immunoglobulin The binding site originate during differentiation of lymphocytes without contact with antigen

6 ALTERNATIVE POSSIBILITIES The GENE CONVERSION is an alternative possibility how to create the high variability of binding site in lymphocyte receptors or immunoglobulins The intrachromosomal gene conversion is the mechanisms in which the transfer of one functional (V) gene from many various pseudogenes take place This mechanism is typical for birds. The combination of VDJ rearrangement and gene conversion take place in various mammals species cattle pig rabbit DIFFERENTIATION OF LYMPHOCYTES LYMPHATIC ORGANS Pluripotent stem cell PRIMARY diferenciation of lymphocytes without the antigen Thymus T lymphocytes Burza fabricii / bone marrow B lymphocytes SECONDARY immune response after contact with antigen NK Lymphoid stem cell T Pro-Ly B CFU-GM Myeloid stem cell CFU-GEMM Pt Ery T c T h Mφ Mo DC Ne Eo Ba MC

7 STRUCTURE OF THYMUS dividing thymocytes apoptotic cell thymocytes nurse cell interdigiting dendritic cell Hassall s body macrophage blood vessel medullar epihelial cell kortical epithelial cell DIFFERENTIATION OF T LYMPHOCYTES nondiferentiated progenitor - migrate into the kortex of the thymus pro-t progenitor - under influence of IL-7 gain marks of T cell pre-t lymphocyte double negative double positive - CD4 - CD8 transcription of genes for CD3 - transcription of genes for TCR - originate γδtcr Ly origin of pre-tcr with β chain - CD4 + CD8 + rearrangement of α chain, origin of TCR with αβ chain positive selection- selection of cells capable to bind to self MHC negative selection - migration to the medulla liquidation of autoreactive clones of cells T lymphocyte - origin of CD4 + CD8 - and CD4 - CD8 + subpopulations (according to the affinity to MHCI or MHCII) DIFERENCIACE T LYMFOCYTŮ DIFFERENTIATION OF B LYMPHOCYTES nondifferentiated progenitor - in contact with reticulum of bone marrow pro-b progenitor - rearrangement of VDJ genes, origin of heavy µ chain pre-b lymphocyte - origin of prebcr with definite heavy and provisional light chain and Igα and Igβ immature B Ly - under influence of IL-3 a IL7 origin of BCR with definite κ or λ chain negative selection - liquidation of autoreactive clones of cells B mature lymphocyte - mature cells with definite BCR enter into secondary lymphatic organs B lymphoblast - after contact with antigen Lymphocytes pass through somatic hypermutation and selection to plasma cell or memory B lymphocyte

8 DIFERENCIACE B LYMFOCYTŮ Specific recognition in immune system is mediated by special protein molecules the only ones which are not created based on genetic matrix, but using random rearrangement of gene segments IMPORTANT PROCESSES DURING DIFFERENTIATION Variability of molecules created by this way is estimated to , what is more than real number of compound in nature Origin of mature lymphocyte from precursors Rearrangement of genes and origin of many clones with different specificity These processes is realized without contact with antigens During negative selection of lymphocyte originate central immunology tolerance (to self antigens) Variability of antibodies is given by the variability of clones of lymphocytes Primary antibody repertoire originate during diferentiation of B lymphocytes (during foetal period) oder sponaneously or after reaction with autoantigens

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