Cross presentation one cell type, pro APCs, can present the antigens of other cells (infected cells)

) and prime/activate nave T-lymphocytes specific for the antigen. MHC class I and II molecules are membrane proteins that each contains a peptidebinding cleft at its amino terminal end. MHC class 1 An alpha chain covalently attached to protein Beta-2 microglobulin. The alpha-1 and alpha-2 domains form the peptide binding cleft. 8-11 amino acids. Alpha-3 is binding site for CD8+ T-cells. MHC class 2 Two chains, alpha and beta. Peptide binding cleft at amino terminal end is at the alpha-1 and beta-1 domains. 10-30 residues. Beta-2 domain contains binding site for CD4+ T-cells. TCR vs. BCR - recognition and signaling molecules Similarities: integral membrane proteins, unique binding sites, made before cell encounters antigen Differences: structure, genes that encode them, epitope they bind Antigen recognizing portions of the receptors are called Variable (V) regions, and conserved portions is Constant (C) regions Complemtarity Determining regions (CDRs) they form the parts of the receptors that bind antigens Collection of antigen receptors and signaling molecules in B lymphocytes is called the B cell receptor Cross-linking adjacent antigen receptors of lymphocytes bind to two or more antigens, signaling cascades Antibodies aka Immunoglobulins(Ig). B-cell membrane bound antigens recognize antigens to initiate humoral reponse, and secreted antibodies elimate antigens in the effector phase of response. Can be membrane bound or secreted proteins, but TCRs only as membrane receptors on T cells. - Composed of four poly-peptide chains, two identical heavy chains, and two identical light chains, each one has 1 variable and 1 constant region. Light 1 V domain, 1 C domain Heavy 1 V domain, and 3-4 C domains. Each variable region contains 3 CDRs, CDR3 has greatest variability, located at junctions of V and C regions. CDR3 contributes most to antigen binding. FAB fragment antigen binding, antigen binding site. Contains a whole light chain (with single V and C domains) attached to the V and first C domains of a heavy chain. Formed by interaction of heavy and light chain variable regions. Rearrangments=DIVERSITY

Fc region remaining heavy chain C domains make up Fc (fragment crystalline) region. Constant region of heavy chain. Class of antibody defined by Fc region. Gives functionality, allows complement (MAC) to become activated Each Ig (antibody) molecule has two identical Fab regions that bind antigens, and one Fc region that is responsible for most biological/effector functions Only difference between BCR of B-cell and single Ig antibody produced is the membrane spanning domain, which retains the BCR on the surface of the B-cell Two types of light chains (Kappa and Lambda). Different C regions, but same functions. Five types of heavy chains (mu, delta, eplison, gamma, alpha) Isotypes/classes antibodies that contain different heavy chains that are named according to their heavy chains, regardless of the light chain class. IgE, IgA, IgG, IgM, IgD. Different effector functions. Antigen receptors of nave B lymphocytes are membrane bound IgM and IgD. Change in Ig isotype prudction called heavy chain class/isotype switching Epitopes/determinants parts of antigens that are recognized by antibodies, could be based on sequence or shape, Affinity strength with which one antigen binding surface of an antibody bind to the epitope. Affinty maturation increase in antigen binding strength (like from secondary infection) Avidity total strength of binding, greater than single affinity bond. Cross reaction antibodies produced for one antigen may bind to other similar antigens. T-cell receptor has an alpha and beta chain, each containing 1 variable and 1 constant region. Binding occurs at alpha and beta variable regions. Like antibodies, each V region contains 3 CDRs, and CDR3 is again most variable. Anchored in plasma membrane, no secreted form, TCRdo not undgo class switching or affinity maturation unlike antibodies. Both chains participate in specific recognition of MHC molecules and bound peptides. TCR/BCR can recognize antigens, but cannot send signals to the cell. TCR- Associated with TCR is complex of proteins called CD3 molecules that wavy chain that make up TCR complex and send some signals when TCR recognizes antigen. T cell activation also requires coreceptor molecules. Maturation of lymphocytes: 1. proliferation of immature cells at several stages, maximizes number of cells that are available to express use antigen receptors. Stimulated by IL-7 produced by stromal cells in bone marrow and thymus. After receptors are expressed, receptors deliever signals for proliferation of only healthy clones. 2. Expression of antigen receptor genes 3. selection of lymphocytes that express useful antigen receptors.

Antigen recptors are produced from several gene segments that are separate from one another in the germile and recombine during lymphocyte maturation. Failure to express pre-lymphocyte OR antigen receptor CELL DEATH Positive selection Immature lymphocyte selected to recognize self MHC molecules. Binds and recognizes MHC molecules, binds weakly GOOD, doesnt bind- dies Negative selection Elimates potentially dangerous lymphocytes that bind to strongly to self MHC molecules, which could react against them. T regs formed in negative selection. Expression of B/T receptors initiated by Somatic Recombination of gene segments that code for the variable regions of the receptors, diversity created Each chain contains V (variable) region genes 200+ (responsible for differences in antibodies), and one or a few Constant (C) regions. Between are small stretches called Diversity (D) and Joining (J) gene segments. All have VJC, only Ig heavy chain and TCR beta chain also contain D segment. Somatic recombo mediated by enzymes called the V(D)J recombinase, the lymphoid specific component of recombinase are RAG-1 and RAG-2 (recombinase activating gene) protiens, brings VDJ segments close together to have DNA cut at end of segments by exonucleases. Then repaired by ligases, forming fully recombined VJ or VDJ gene. TdT takes nucleotides that are not parts of germline and adds these nucleotides randomly to the VDJ segments. (forming N regions). All this can also form nonfunctional genes, which is why there are several checkpoints. Maturation of B-cells occurs mainly in the bone marrow.. 1. Progenitors of B-cell lineage proliferate under influence of IL-7 giving B-cell precursors called PRO-B cells. The next stage Mu-heavy chain formed called PRE-B cells. Some of the Mu protein is expressed on the surface to form pre-BCR. First checkpoint, makes sure Mu protein is produced and functional, if not, DEATH. Selects and expands cells with functional Mu heavy chain. 2. This signals other processes, which shuts of recombo on the 2nd chromosome, and the other signal triggers recombo of light chain, first kappa then Lamda. Whichever light chain produced associated with Mu-heavy chain to complete IgM antigen receptor. This receptor delievers signals for proliferation for cells with COMPLETE receptors, 2nd checkpoint. IgM expressing B lymphocyte is IMMATURE B-cell. 3. Further maturation may occur in bone marrow or peripheral tissues. Final step, coexpression of IgD with IgM, because recombined VDJ heavy chain may be spliced into RNAmRNA, IgM+ IgD+ cell is MATURE B-cell can respond to antigen.

B-cells go through positive and negative selection. Removes potentially dangerous cells, if they bind too tight (negative selection) to self-molecules in bone marrow. Looks for intact receptors (positive selection) B-cell diversity - 1. Somatic Recombination (combinatorial diversity) 2. VDJ nucleotide alteration (Junctional Diversity) 3. Different Heavy and Light chain combos 4. Somatic Hypermutation T-cell genetics - VDJ segment is spliced onto constant region. V reapted 50 times, J repeated 12-60 times. NO SOMATIC HYPERMUTATION Junctional Diversity - adding or deleting nucleotides, adding nucleotides forms N regions. Overhanging DNA filled in by P nucleotides. T-cells - 1. Stem cellsDouble negative (CD8-CD4-) PRO-T-cell- Pre-T-cellDouble positive immature cell Negative and Positive selection, weak bind to MHC class I becomes CD8+, weak bind to MHC class II becomes CD4+. Complete TCR at double positive/immature cell stage Tolerance - unresponsiveness to self-antigens, can tell what is self and non-self. Failure can lead to autoimmume disease. When antigen and antigen specific lymphocyte come in contact: 1. Activation proliferation/diffy caused by immunogenic antigen 2. Tolerance - functionally inactivated or killed (self antigen) 3. Ignorance - does not react at all to self antigen Central Tolerance - at bone marrow or thymus. When lymphocytes encounter selfantigens. T cells- anergy - lack of response, not enough costimulators. Negative selection. B cells- Negative selction or receptor editing -changes light chain. Peripheral - in secondary tissues. B-cells-just anergy

T lymphcytes have to interact with other cells, phagocytes, infected cells, B lymphs, in order to perform their functions. Differentiation - conversion of nave T cells to effector cells. TCR and either CD4 or CD8 CO-RECEPTOR recognize antigens on MHC molecules, and function with the TCR to bind MHC molecules. TCR complex "signaling complex" = TCR, CD3, and wavy chain Adhesion molecules - stabilize binding of T cells to APCs. Main ones are INTEGRINS LFA-1 ON T-cell binds to ICAM-1 on APC. Low affinity at first until exposed to chemokines then high affinity state and LFA receptors cluster together to bind strongly.

Costimulators - CD28 receptor on T-cells binds to B7-1/2, initiates responses in nave T cells. SECOND SIGNAL Others: CD40L on T-cell, CD-40 on APC, activates APC to express more B7 and activate cytokines, IL-2, which enhances T-cell diffy. Makes APCs better (CD-40) CD8+ (CTL, kills cells themselves) must have co-activation of CD4+ (mainly activate macrophages and B lymphocytes) CD4+ can either be Th1 or Th2. Th1 - produces INF-gamma, TNF. activated macrophages. IL-12 (from macrophages) causes Th1 subset. Primarily cytotoxic, macrophage activation. If no IL-12, then cells themselves prodce IL-4, which causes Th2 subset. Th2 - From IL-4. produces IL-4, stimulated IgE antibodies, and IL-5, activates eosinophils. Also produces IL-4, IL-10, IL-13 which inhibit macrophage activation and suppress Th1 cells. Tregs - produce IL-10, IL-13, TGF-Beta to down regulate immune system after infection is gone. Have protein called CD25. CTL/CD8+ activation - binds to APC that has ingested antigen, either cytokines from self or from helper T cell. T cells then release perforin, which put holes in membrane, then granzymes are bombs that kills cell. Granzyme B activates caspace. CD4/CD8 co-receptors - activates protein called Lck attached to tails, binds with ITAMs on CD3 and wavy chains. This becomes docking site for ZAP-70, this starts activation of signaling. After that PLC IP3 or DAG- calcineriun or PKC- NFAT. All this starts activation of CD-40L and cytokines, etc. Nave =T cells stay in lymph node though L-selectin binding. Effectors find by cytokines and stay in place by E/P selectins, OR ICAM-1/VCAM-1 at infection site. ICAM1LFA-1 VCAM-1VLA-4 If T-cell recognizes antigen, increases binding affinity of VLA-4, which can bind to extracellular matrix