T cell maturation, activation and differentiation

T lymphocytes
Activates immune and inflammatory response lymphokine

It expresses different membrane molecules:

Thy-1: earliest marker of T cell lineage, remains throughout life TCR: present on all cells; two polypeptide chain CD3: six polypeptide chain complex associated with TCR CD4 or CD8: CD4 on TH cells; CD8 on TC cells CD28: ligand for co stimullatory B7 molecules present on APC CD45: participate in signal transductions

 After the naive T cell (N) encounters an antigen - proliferate (divide) into many clones or daughter cells. Some will differentiate into effectors T cells

(E) that will produce cytokines

Some will form memory T cells (M) that will survive in an inactive state

T lymphocytes
Differ from B cell and NK cell by having TCR Earliest thymocytes express neither CD4 nor CD8, and are therefore classed as double-negative (CD4-CD8-) cells Double negative double positive single positive released from the thymus to peripheral tissues. About 98% of thymocytes die during the development processes in the thymus Either by failing positive selection or negative selection,

Positive selection
"selects for" T-cells capable of interacting with MHC Double-positive thymocytes (CD4+/CD8+) move deep into the thymic cortex where they are presented with self-antigens complexed with MHC molecules on the surface of cortical epithelial cells. Only those thymocytes that bind the MHC/antigen complex with adequate affinity will receive a vital "survival signal. Because of this, the thymocytes with no affinity for self antigens die by apoptosis and are engulfed by macrophages. Double-positive cells (CD4+/CD8+) that are positively selected on MHC class II molecules will eventually become CD4+ cells, while cells positively selected on MHC class I molecules mature into CD8+ cells.

 Removes thymocytes that are capable of strongly binding with "self" peptides presented by MHC. Remove thymocytes that may cause autoimmunity Thymocytes that survive positive selection migrate towards the boundary of the thymic cortex and thymic medulla. While in the medulla, they are again presented with selfantigen in complex with MHC molecules on antigenpresenting cells (APCs) such as dendritic cells and macrophages. Thymocytes that interact too strongly with the antigen receive an apoptotic signal that leads to cell death.

Negative selection

Positive selection selects for T cells that are capable of recognizing self antigens through MHC. Negative selection selects for T cells that bind too strongly to self antigens.

 Responsible for recognizing antigens bound to MHC Binding between TCR and antigen is of relatively low affinity and is degenerate Heterodimer; in 95% of T cells contains alpha () and beta () fragment, whereas in 5% of T cells has gamma and delta (/) fragment Transmembrane region of the TCR is composed of positively charged amino acids; such structure allows the TCR to associate with other molecules like CD3, which possess three distinct chains (, , and ) or 2 complex or a / complex These accessory molecules have negatively charged transmembrane regions and are vital to propagating the signal from the TCR into the cell; The cytoplasmic tail of the TCR is extremely short, making it unlikely to participate in signaling.

T cell receptors (TCR)

Helper T cell (TH Cells)

Assist other white blood cells in immunologic processes They have no cytotoxic or phagocytic activity; they cannot kill infected host cells or pathogens They are essential in determining B cell antibody class switching, in the activation and growth of cytotoxic T cells, and in maximizing bactericidal activity of phagocytes such as macrophages. For proliferation they need IL2, secreted by itself (autocrine)

they express the T cell receptor-CD3 complexS

 After many cell generations, the Th cell's progenitors differentiate into effector Th cells, memory Th cells, and regulatory Th cells.

Effector Th cells secrete cytokines, proteins or peptides that stimulate or interact with other leukocytes, including Thcells. Memory Th cells retain the antigen affinity of the originally activated T cell, and are used to act as later effector cells during a second immune response (e.g. if there is re-infection of the host at a later stage). Regulatory T cells do not promote immune function, but act to decrease it instead. Despite their low numbers during an infection, these cells are believed to play an important role in the self-limitation of the immune system; they have been shown to prevent the development of various auto-immune diseases.

 T-Killer cell, cytolytic T cell, CD8+ T-cells or killer T cell) Inducing the death of infected somatic or tumor cells Virus-infected cells (e.G., HIV-infected CD4+ T cells); Cells infected with intracellular bacterial or protozoal parasites; Allograft such as transplanted kidney, heart, lungs, etc. Two types of killing Perforin/Granzyme Killing perforin molecules insert themselves into the plasma membrane of target cells forming a pore that enables Granzymes are serine proteases. The two most abundant ones are-- Granzyme A. Once inside the cell, it cleaves a subunit of complex I (the NADH dehydrogenase) of the ETC producing ROS that kill the cell. Granzyme B. Once inside the cell, it proceeds to cleave the precursors of caspases thus activating them to cause the cell to self-destruct by apoptosis.

Cytotoxic T cell (Tc Cells)

 FasL/Fas Killing Express on their surface molecules of a transmembrane protein, the death activator designated Fas ligand (FasL) (Apo1L)(CD95L) Most potential CTL targets express a receptor for FasL designated Fas (Apo1)(CD95) When cytotoxic T cells recognize (bind to) their target, they produce more FasL at their surface. This binds with the Fas on the surface of the target cell leading to its death by apoptosis.

Memory T cell (TM Cells)

Types
Central memory TCM cells express L-selectin (CD62L) and the CCR7, they secrete IL-2, but not IFN or IL-4. Effector memory TEM cells, however, do not express Lselectin or CCR7 but produce effector cytokines like IFN and IL-4. Explored using co-stimulatory molecules CD27 and CD28 expression in addition to CCR7 and CD62L

Natural Killer T cells

Share properties of both T cells and natural killer (NK) cells Bridges between innate between adaptive immunity, Co-express an T cell receptor (TCR), but also express a variety of molecular markers that are typically associated with NK cells, such as NK1.1. Share other features with NK cells as well, such as CD16 and CD56 expression and granzyme production. Unlike conventional T cells that recognize peptide antigen presented by MHC molecules, NKT cells recognize glycolipid antigen presented by a molecule called CD1d, They are also able to recognize and eliminate some tumor cells and cells infected with herpes viruses,

 Once activated, these cells can perform functions ascribed to both TH and TC cells (i.e., cytokine production and release of cytolytic/cell killing molecules), Upon activation, NKT cells are able to produce large quantities of interferon-gamma, IL-4, and granulocyte-macrophage colonystimulating factor, as well as multiple other cytokines and chemokines (such as IL-2, Interleukin-13, Interleukin-17, Interleukin-21 and TNF-alpha).

 Specialised antigen presenting cells are primarily dendritic cells, macrophages and B cells, although B cells are the only cell group that expresses MHC Class II constitutively (at all times). Some APCs also bind native (or unprocessed) antigens to their surface, such as follicular dendritic cells, but unprocessed antigens do not interact with T cells and are not involved in their activation. The antigens that bind to MHC proteins are always short peptides, 8-10 amino acids long for MHC Class I, and up to 25 or so for MHC Class II.

Signal 1 of T cell activation

Endocytose (absorb) foreign material, which undergoes processing, then travels from the infection site to the lymph nodes.

APC begins to present antigen peptides with Class II MHC, allowing CD4+ T cells that express the specific TCRs.
TCR-CD3 complex binds strongly to the peptide-MHC complex. CD4, also binds to a different section of the MHC molecule. These interactions bring these proteins closer together, allowing the intracellular kinases present on the TCR, CD3 and CD4 proteins to activate each other via phosphorylation. With the assistance of a phosphatase present on the intracellular section of CD45 (common leukocyte antigen), these molecules activate the major biochemical pathways in the cytosol of the Th cell.

 The integrin protein LFA-1 on the T cell and ICAM on the APC are the primary molecules of adhesion in this cell interaction.

Involves an interaction between CD28 on the CD4+ T cell and the proteins CD80 (B7.1) or CD86 (B7.2) on the professional APCs. Both CD80 and CD86 activate the CD28 receptor. Once the two-signal activation is complete the T helper cell (Th) then allows itself to proliferate by releasing a potent T cell growth factor called interleukin 2 (IL-2) which acts in an autocrine fashion. Activated T cells also produce the alpha sub-unit of the IL-2 receptor (CD25 or IL-2R. The Th cells receiving both signals of activation will then become Th0 cells (T helper 0) cell that secrete IL-2, IL4 and interferon gamma (IFN-).

Verification (Signal 2)

 The Th0 cells will then differentiate into Th1 or Th2 cells depending on cytokine environment. IFN- drives Th1 cell production while IL-10 and IL-4 inhibit Th1 cell production. Conversely, IL-4 drives Th2 cell production and IFN- inhibits Th2 cells.

TheTh0 cells are nave T cells that produce cytokines typical of both Th1 and Th2 cells. In general,Th1cells produce lymphokines that stimulate macrophages and cytotoxic Tcells whilstTh2 cells produce lymphokines that stimulate B cells to proliferate and produce antibody. In contrast, Th3 cells produce cytokines that are involved in the regulation or switching off of the immune response.

Th1 Main partner cells Cytokines produced Immune stimulation promoted Macrophage IFN-, TNF Cellular immune system. Maximizes the killing efficacy of the macrophages Increases the production of interleukin-12 by dendritic cells and macrophages.

Th2 B cells IL4, IL5, IL6, IL10, IL13 Humoral immune system. Stimulates B-cells into proliferation, to induce Bcell antibody class switching. IL4 acts on helper T cells to promote the production of Th2 cytokines whileIL-10 inhibits IL2 and IFN- in helper T cells and IL-12 in dendritic cells and macrophages.

Other function

 B cells are produced in the bone marrow. have immunoglobulin and other molecules (e.g. CD19, 20, 23, 24, 35 and 40) inserted in their membranes.

B cells

Transmembrane receptor protein located on the outer surface of B-cells In addition to Immunoglobulin (mIg), the BCR comprises a duplex of molecules known as CD79a/b (formerly Ig and Ig) that have a single extracellular region and a cytoplasmic tail.

BCR

The B-cell receptor is composed of two parts

Ligand binding moiety Signal transduction moiety: heterodimer called Ig-/Ig- (CD7. Each member of the dimer spans the plasma membrane and has a cytoplasmic tail bearing an immunoreceptor tyrosine-based activation motif (ITAM))

 Both BCRs and TCRs share these properties: They are integral membrane proteins. They are present in thousands of identical copies exposed at the cell surface. They are made before the cell ever encounters an antigen. They are encoded by genes assembled by the recombination of segments of DNA. They have a unique binding site. This site binds to a portion of the antigen called an antigenic determinant or epitope. The binding, like that between an enzyme and its substrate depends on complementarity of the surface of the receptor and the surface of the epitope. The binding occurs by non-covalent forces (again, like an enzyme binding to its substrate).

BCR and TCR

 BCRs and TCRs differ in:

Their structure; The genes that encode them; The type of epitope to which they bind.

Major histocompatibility complex

MHC complex is group of genes on a single chromosome that codes the MHC antigens MHC molecules play an important role in the immune system and autoimmunity. MHC genes present proteins through several mechanisms:
the MHC locus is polygenic, MHC genes are highly polymorphic and numerous alleles have been described, and several MHC genes are codominantly expressed.

MHC1
Composed of two polypeptide chains, a long chain and a short chain called 2-microglobulin The chain has four regions.

The 2- microglobulin associates with the chain and helps maintain the proper conformation of the molecule.

A cytoplasmic region, containing sites for phosphoylation and binding to cytoskeletal elements. A transmembrane region containing hydrophic amino acids by which the molecule is anchored in the cell membrane. A highly conserved 3 immunoglubilin-like domain to which CD8 binds. A highly polymorphic peptide binding region formed from the 1 and 2 domains.

Class II MHC molecules are composed of two polypeptide chains an and a chain of approximately equal length. Both chains have four regions:
A cytoplasmic region containing sites for phosphoylation and binding to cytoskeletal elements

MHC2

A transmembrane region containing hydrophic amino acids by

which the molecule is anchored in the cell membrane A highly conserved 2 domain and a highly conserved 2 domain

to which CD4 binds

A highly polymorphic peptide binding region formed from the 1 and 1 domains

 Each MHC molecule has only one binding site. The different peptides a given MHC molecule can bind all TLC to the same site, but only one at a time. Because each MHC molecule can bind many different peptides, binding is termed degenerate. MHC polymorphism is determined only in the germline. There are no recombinational mechanisms for generating diversity. MHC molecules are membrane-bound; recognition by T cells requires cellcell contact. Alleles for MHC genes are co-dominant. Each MHC gene product is expressed on the cell surface of an individual nucleated cell. A peptide must associate with a given MHC of that individual, otherwise no immune response can occur. That is one level of control. Mature T cells must have a T cell receptor that recognizes the peptide associated with MHC. This is the second level of control. Cytokines (especially interferon-) increase level of expression of MHC. Peptides from the cytosol associate with class I MHC and are recognized by Tc cells. Peptides from within vesicles associate with class II MHC and are recognized by Th cells. Polymorphism in MHC is important for survival of the species.

Name

Expression All nucleated cells. MHC class I proteins Encodes non-identical pairs contain an chain & 2-micro-globulin (heterodimer) of peptide(not part of the MHC encoded by binding proteins, as well chromosome 15). They present antigen MHC class I as antigen-processing fragments to cytotoxic T-cells via molecules such as TAP and the CD8 receptor on the cytotoxic TTapasin. cells and also bind inhibitory receptors on NK cells. Encodes (1) heterodimeric peptide-binding proteins and On most immune system cells, specifically (2) proteins that modulate on antigen-presenting cells. MHC antigen loading onto the class II proteins contain & chains MHC class II peptideMHC class II and they present antigen fragments to binding proteins in T-helper cells by binding to the lysosomal compartment the CD4 receptor on the T-helper such as MHC II DM, MHC II cells.

the 3.6-Mb MHC region on chromosome 6 contains 140 genes

Function

 In human HLA; in rat H2 complex MHC III-

components of the complement system (such as C2, C4 and B factor) and molecules related with inflammation (cytokines such as TNF-, LTA, LTB) or heat shock proteins (hsp)

 Class I MHC molecules bind peptides derived from endogenous antigens that have been processed within the cytoplasm of the cell (e.g., normal cellular proteins, tumor proteins, or viral and bacterial proteins produced within infected cells). Class II MHC molecules bind peptides derived from exogenous antigens that are internalized by phagocytosis or endocytosis and processed within the endocytic pathway.

Characteristics of the antigen processing pathways

Characteristic MHC-I pathway MHC-II pathway Polymorphic chain and 2 Composition of the stable peptidePolymorphic chains and , microglobulin, peptide bound to MHC complex peptide binds to both chain Types of antigen presenting cells (APC) Dendritic cells, mononuclear phagocytes, B lymphocytes, some endothelial cells, epithelium of thymus Helper T lymphocytes (CD4+) Proteins present in endosomes or lysosomes (mostly internalized from extracellular medium) Proteases from endosomes and lysosomes (for instance, cathepsin)

All nucleated cells, platelets

T lymphocytes able to respond

Cytotoxic T lymphocytes (CD8+) cytosolic proteins (mostly synthetized by the cell; may also enter from the extracellular medium via phagosomes)

Origin of antigenic proteins

Enzymes responsible for peptide Cytosolic proteasome generation

Location of loading the peptide on Endoplasmic reticulum the MHC molecule Molecules implicated in transporting the peptides and TAP (transporter associated with loading them on the MHC antigen processing) molecules

Specialized vesicular compartment

DM, invariant chain

Transporter associated with antigen processing (TAP)

member of the ATP-binding-cassette transporter family formed of two proteins: TAP-1 and TAP-2, which have one hydrophobic region and one ATP-binding region each found in the ER lumen associated with the peptide-loading complex (PLC) consist of 2 microglobulin, calreticulin, ERp57, TAP, tapasin, and MHC class I Both nucleotide-binding domains (NBDs) are required for peptide translocation, as each NBD cannot hydrolyse ATP alone. ATP binding to TAP-1 is the initial step in the transport process, and that ATP bound to TAP-1 induces ATP binding in TAP-2.

 Both TAP1 and TAP2 belong to the family of ATP-binding cassette proteins found in the membranes of many cells, including bacteria; these proteins mediate ATP-dependent transport of amino acids, sugars, ions, and peptides. Peptides generated in the cytosol by the proteasome are translocated by TAP into the RER by a process that requires the hydrolysis of ATP TAP has the highest affinity for peptides containing 810 amino acids, which is the optimal peptide length for class I MHC binding. TAP appears to favor peptides with hydrophobic or basic carboxyl-terminal amino acids, the preferred anchor residues for class I MHC molecules. TAP is optimized to transport peptides that will interact with class I MHC molecules.

 The TAP1 and TAP2 genes map within the class II MHC region, adjacent to the LMP2 and LMP7 genes Both the transporter genes and these LMP genes are polymorphic; that is, different allelic forms of these genes exist within the population TAP deficiencies can lead to a disease syndrome that has aspects of both immunodeficiency and autoimmunity Within the RER membrane, a newly synthesized class I chain associates with calnexin until 2-microglobulin binds to the chain. The class I chain/2-microglobulin heterodimer then binds to calreticulin and the TAP-associated protein tapasin. When a peptide delivered by TAP is bound to the class I molecule, folding of MHC class I is complete and it is released from the RER and transported through the Golgi to the surface of the cell.

 The endocytic pathway appears to involve three increasingly acidic compartments: early endosomes (pH 6.06.5); late endosomes, or endolysosomes (pH 5.06.0); and lysosomes (pH 4.55.0). Lysosomes, for example, contain a unique collection of more than 40 acid-dependent hydrolases, including proteases, nucleases, glycosidases, lipases, phospholipases, and phosphatases.

 Longer peptides bulge in the middle, whereas shorter peptides are more extended. Contact with the MHC molecule is by hydrogen bonds to anchor residues 1/2 and 8/9.

human cytomegalovirus (CMV), hepatitis B virus (HBV), and adenovirus 12 (Ad12) decreases the expression of MHC 1.

 These molecules consist of two, linked, glycoproteins ( and ), composed of 229 and 237 amino acids, respectively Each is coded for by genes of the MHC and consists of four regions; two extracellular, hydrophilic regions (1 and 2, or 1 and 2 domains), a transmembraneous, hydrophobic region; and an intracellular, hydrophilic, region that anchors the molecule in the cell membrane. 1 and 1 domains combine to form a single peptidebinding site composed of two a helical loops supported by a platform of eight anti-parallel b strands

MHCII

 Recent experiments indicate that most class II MHCinvariant chain complexes are transported from the RER, where they are formed, through the Golgi complex and trans-Golgi network, and then through the endocytic pathway, moving from early endosomes to late endosomes, and finally to lysosomes. A short fragment of the invariant chain termed CLIP (fclass II associated invariant chain peptide) remains bound to the class II molecule after the invariant chain has been cleaved within the endosomal compartment. CLIP physically occupies the peptide-binding groove of the class II MHC molecule, presumably preventing any premature binding of antigenic peptide HLA-DM, a nonclassical MHC class II molecule expressed within endosomal compartments, mediates exchange of antigenic peptides for CLIP. The nonclassical class II molecule HLA-DO may act as a negative regulator of class II antigen processing by binding to HLA-DM and inhibiting its role in the dissociation of CLIP from class II molecules.

 HLA-DM is widely conserved among mammalian species. Like other class II MHC molecules, HLA-DM is a heterodimer of and chains. Unlike other class II molecules, HLA-DM is not polymorphic and is not expressed at the cell membrane but is found predominantly within the endosomal compartment.

 HLA-DO differs from HLA-DM in that it is expressed only by B cells and the thymus, and unlike other class II molecules, its expression is not induced by IFN-. The antigen-binding groove is open at both ends and does not enclose the C- and N-termini of the peptide. The bound peptide extends beyond the antigen-binding groove at both ends.

Role of MHC molecules in transplant rejection

Each human cell expresses six MHC class-I alleles (one HLA-A, -B, and -C allele from each progenitor) and 6-8 MHC class-2 alleles (one HLA-DP and -DQ, and one or two HLA-DR from each progenitor, and some combinations of these). The MHC polymorphism is very high: It is estimated that in the population there are at least 350 alleles for HLA-A genes, 620 alleles for HLA-B, 400 alleles for DR, and 90 alleles for DQ All MHC molecules can be targets for transplant rejection, but HLA-C and HLA-DP molecules show low polymorphism, and most likely they are less important in rejection.

B cell maturation, activation and differentiation

 Activation of CD4+ T cells occurs through the engagement of both the T cell receptor and CD28 on the T cell by the major histocompatibility complex (MHC) peptide and B7 family members on the APC, respectively. Absense of co-stimullator CD28 - Anergy

Activation

 The immunoglobulin molecule consists of three major functional regions called the

Immunoglobulin

The F(ab)2 region (Fragment antigen binding) contains the binding sites for foreign particles (or antigens), the hinge region is where the other two meet and confers flexibility on the molecule and Fc region (Fragment crystallisable) confers the biological properties of the molecule

The classic signs and symptoms of acute inflammation

English
Redness Swelling Heat Pain

Latin
Rumor Tumor Calor Dolor Celsus

Loss of function

Functio laesa

Galen

Comparison between acute and chronic inflammation

Acute
Causative agent Pathogens, injured tissues Neutrophils, mononuclear cells (monocytes, macrophages) Vasoactive amines, eicosanoids Immediate Few days Resolution, abscess formation, chronic inflammation

Chronic Persistent acute inflammation due to non-degradable pathogens, persistent foreign bodies, or autoimmune reactions
Mononuclear cells (monocytes, macrophages, lymphocytes, plasma cells), fibroblasts IFN- and other cytokines, growth factors, reactive oxygen species, hydrolytic enzymes Delayed Up to many months, or years Tissue destruction, fibrosis, necrosis

Major cells involved

Primary mediators Onset Duration Outcomes

Plasma cascade systems

The complement system, when activated, results in the increased removal of pathogens via opsonisation and phagocytosis. The kinin system generates proteins capable of sustaining vasodilatation and other physical inflammatory effects. The coagulation system or clotting cascade which forms a protective protein mesh over sites of injury. The fibrinolysis system, which acts in opposition to the coagulation system, to counterbalance clotting and generate several other inflammatory mediators.

Plasma derived mediators

Name Bradykinin C3 Produced by Kinin system Complement system Description
A vasoactive protein which is able to induce vasodilation, increase vascular permeability, cause smooth muscle contraction, and induce pain. Cleaves to produce C3a and C3b. C3a stimulates histamine release by mast cells, thereby producing vasodilation. C3b is able to bind to bacterial cell walls and act as an opsonin, which marks the invader as a target for phagocytosis. Stimulates histamine release by mast cells, thereby producing vasodilation. It is also able to act as a chemoattractant to direct cells via chemotaxis to the site of inflammation. A protein which circulates inactively, until activated by collagen, platelets, or exposed basement membranes via conformational change. When activated, it in turn is able to activate three plasma systems involved in inflammation: the kinin system, fibrinolysis system, and coagulation system. A complex of the complement proteins C5b, C6, C7, C8, and multiple units of C9. The combination and activation of this range of complement proteins forms the membrane attack complex, which is able to insert into bacterial cell walls and causes cell lysis with ensuing death. Able to break down fibrin clots, cleave complement protein C3, and activate Factor XII. Cleaves the soluble plasma protein fibrinogen to produce insoluble fibrin, which aggregates to form a blood clot. Thrombin can also bind to cells via the PAR1 receptor to trigger several other inflammatory responses, such as production of chemokines and nitric oxide.

C5a
Factor XII

Complement system
Liver

Membrane attack complex Plasmin Thrombin

Complement system

Fibrinolysis system Coagulation system