Immuno. Chapter 8

Secreted cytokines

+Secreted cytokinesand related membrane-bound proteins that act through cell-surface receptors generally to induce changes in gene expression within target cells. Work locally and work over short period of time -Autocrines -Paracrines Colony stimulating factors: work at a distance: stimulate bone marrow +Many cytokines made by T cells are called interleukins (numbered in order of discovery, IL-2 or IL-7). +Cytokines (general term) made by lymphocytes are called lymphokines (more specific term).

CTLA-4

-Activated T cells express another B7 receptor called _______. -It binds B7 twentyfold more strongly than does CD28 and functions as an antagonist. -So B7 binding to CD28 activates a T cell, while B7 binding to ______ slows down activation and limits cell proliferation.

Cytokines

-Alter the behavior of target cells. -All effector T cells produce cytokines (just different types and in different combinations).

Cell-surface molecules of the immunoglobulin superfamily initiate lymphocyte adhesion to APC

-As naïve T cells move through the cortex of the lymph node, these will bind transiently to APCs. -The initial encounter of T cells with antigen-presenting dendritic cells involves integrins and members of the immunoglobulin superfamily: The T cell's LFA-1 binds to ICAM-1 or ICAM-2 on the APC The APC's LFA-1 binds to ICAM-3 on the T cell =Adhesion is strengthened by:CD2 on the T cell binding to LFA-3 on the APC. ICAM-3 on the T cell and the lectin DC-SIGN on activated dendritic cell. -These transitory interactions allow the T cell to screen the peptide:MHC complexes on the APCs.

Apoptosis or programmed cell death

-Cell suicide in which cells shrivel or shrink but remain intact. -_______ of the target cell is induced by the cytotoxic CD8 Tcells releasing cytotoxins. Target cells die by it which prevents pathogen replication as well as the release of the infectious pathogen.

TH2

-Defense against parasites in tissues and surfaces -Does not lead to inflammation -Induce IgE production from B-cells +Bind to mast cells, basophils and activated eosinophils -Activated by IL-4 environment, this leads to activation of TF GATA3---> differentiation into TH2 effector cell

CD4 T cells: TH17

-Enhance the neutrophil response to fungal and extracellular bacterial infections -Cytokines-IL-17, IL-21, IL-22 and IL-26

Resting naïve T cells

-Express L- selectin (for homing to LN) and relatively low levels of CD2 & LFA-1 (lymphocyte function -associated antigen-1 on surface of lymphocytes) adhesion molecules -When activated, L-selectin expression stops & increased amounts of integrin LFA-1 are made

Activated Effector T cells

-Express more adhesion molecules than naïve T cells -Express two to four times more adhesion molecules - CD2 and LFA-1 - than naïve cells. -Can interact with target cells expressing lowerlevels of ICAM-1 and LFA-3. -Interactions of effector T cell with target cells is short-lived UNLESS the TCR is engaged by specific antigen. -The interaction leads to a conformational change in LFA-1 that strengthens the adhesion between the two cells.

CD4 T cells: TFH

-Help B cells become activated, switch isotype and increase antibody affinity -Cytiokines-IL-21, Il-4 and IFN-y

CD4 T cells: TH2

-Help basophils, mast cells, esinophils and B cells respond to parasite infections Cytokines-IL-4, IL-5, IL-10, IL-13 and TGF-B

CD4 T Cells: TH1

-Help macrophages to suppress intracellular infections -Cytokines-IFN-y, GM-CSF, LT, and IL-2

TFH

-Help naïve B-cells in follicles/germinal centers become plasma cells or memory cells -Expresses CXCR5 binds to CXCL13 TFH travels to follicle B cell isotype switching

TH1 Cells

-IL-12 and IFN-y are the chracteristic cytokines and the cytokines that induce differentiation -T-bet is the defining transcription factor -Function: Activates macrophages

TFH Cells

-IL-16, TGF-B and IL-23 are the cytokines that induce differentiation -Bcl6 is the defining transcription factor -The characteristic cytokine is IL-21 -Function: Activates B cells Maturation of antibody response

TH2 Cells

-IL-4 is the cytokine that induces differentiation -GATA3 is the defining transcription factor -The characteristic cytokine is IL-21 -Function: Activates cellular and antibody response to parasites

TH17 cells

-IL-6 and IL-21 are the cytokines that induce differentiation -RORyT is the defining transcription factor -The characteristic cytokines are IL-17 and IL-6 -Function: Enhance Neutrophil response

Example of Activated CD4 T cell

-IntegrinVLA-4 is expressed (homing receptor for vascular endothelium at sites of inflammation) and guides activated Tcells to the infection site. -Activated T cells have more surface CD2 which leads to increased adhesion to target cells.

TH1 cells coordinate the host response to intravesicular pathogens

-Intracellular pathogens can enjoy a protected life in the vesicular system of macrophages (tuberculosis and leprosy). -Intravesicular pathogens: +Can not be reached by antibody, nor can their peptides be presented by MHC class I molecules to CD8 T cells. +Avoid digestion by lysosomal enzymes by preventing the acidification of the phagolysosome that is required to activate the lysosomal hydrolases -Infections of this type are fought by TH1 CD4 T cells (that help themacrophages become activated so intracellular pathogens are killed).

CD8 T-Cells:Cytotoxic

-Kill virus infected cells -Cytokines- IFN-y, LT, IL-2 -Cytotoxins-perforin, granzymes, granulysin and serglycin

Naïve T and B lymphocytes circulate in the blood and enter lymph nodes by crossing high endothelial venules

-Lymphocytes bind to the endothelium in the lymph node through interaction of L-selectin with vascular addressins. -Chemokines, which are also bound to the endothelium, activate the integrin LFA-1 on the lymphocyte surface enabling it to bind tightly to ICAM-1 on the endothelial cell. -Establishment of tight binding allows the lymphocyte to squeeze between two endothelial cells, leaving the lumen of the blood vessel and entering the lymph node -The T-cells will migrate toward the high concentrations of CCL21 and CCL19 within the lymph node

Activated T cells secrete and respond to IL-2

-Naïve T cells express the low-affinity receptor for IL-2 which consists of B and y chains -Activation of a naïve T cell by the recognition of a peptide:MHC complex accompanied by co-stimulation induces the synthesis and secretion of IL-2 and the synthesis of the IL-2 receptor a chain -Cell enters the first phase (G1) of cell-division cycle -a chain combines with the B and y chains to make a high-affinity receptor for IL-2 -IL-2 binds to the IL-2 receptor producing an intracellular signal that promotes T-cell proliferation

Once Ag-specific T cells are trapped in the LN by an APC....

-Naïve T cells specific for an antigen are about 1 in 104 or 1 in 106 of the total T cell pool. -Most T cells that enter a lymph node do not encounter a specific antigen and will recirculate for many years. -Once an antigen-specific T cell is trapped in the lymph node by an APC and activated it takes several days for the activated T cell to proliferate and differentiate into effector Tcells . -= delay between onset of infection and appearance of primary adaptive immune response.

Recirculating, Naïve T cells first encounter antigen on APCs in secondary lymphoid tissues

-Naïve T cellsenter lymphoid tissues via blood capillaries or from the afferent lymph. -T cells in blood: Bind to endothelial cells of the thin-walled high endothelial venules (HEV). Squeeze through vessel wall and enter the cortical regionof the lymph node. Encounter dendritic cells and use its TCR to examine the peptide:MHC complexes on dendritic surfaces. -When the T cell encounters a peptide:MHC complex that it can bind to, the T cell is retained in the lymph node and is activated, then proliferates and differentiates into a clone of effector T cells.

Homing of naïve T cells to secondary lymphoid tissues is determined by cell adhesion molecules.

-Passage of naïve T cell out of the bloodstream, through HEV to the lymph node cortex is controlled by cell-surface molecules on the T cells and endothelial cells. -These contacts are initiated by adhesion molecules on the T cell which bind to complementary molecules on other cells. -Adhesion molecules of the immune system comprise four structural classes. -Drawn into the secondary lymphoid tissue by chemokines that bind to their surface receptors. +Receptor on T-cell - CCR7 +Chemokines produced in lymphoid tissue - CCL21 and CCL19

Treg

-Primary cells involved in regulating the immune reponse. -Interact with cells to down regulate their activity -In presence of TGFB the naïve T cell activates TF FoxP3 ---> Treg cell -Produce IL-10 and TGFB Reduce inflammation and immune response -Natural (nTreg) and induced (iTreg) T regulatory cells. nTreg - differentiate in thymus iTreg - differentiate in periphery

TH17

-Recruits neutrophils to site of infection -Secrete IL-17 which induces production of CXCL8 from epithelial cells and stromal cells -TGF-B and IL-21 or IL-6 activate TF RORyT --->differentiation into TH17 effector

Regulatory CD4 T cells limit the activities of effector CD4 and CD8 T cells

-Regulatory or suppressor CD4 T cells make inhibitory cytokines. +IL-4, IL-10, TGF-B with high expression of CD25, and the a chain of IL-2 receptor +Inhibition depends on physical contact between the regulatory CD4 T cell and its target +FoxP3 TF

CD4 T Cells: Treg

-Suppress the activities of other effector T-cell populations -Cytokines-TGF-B, IL-10, IL-35

T regulatory Cells

-TGF-B is the cytokine that induces differentiation -FoxP3 is the defining transcription factor -The characteristic cytokines are IL-10 and TGF-b -Function: Suppress other effector T cells

Naïve CD8 T cells can be activated in different ways to become cytotoxic cells

-The activation of naïve CD8 T cells requires stronger co-stimulation than CD4 T cells. -CD8 T cells stimulated by antigen and co-stimulation will synthesize IL-2 and the high affinity IL-2 receptor, which induces their proliferation and differentiation. -Under conditions of suboptimal co-stimulation, CD4 T cells can help activated naïve CD8 T cells. -CD4 and CD8 T cells must recognize the their specific antigen on the same APC, then the CD4 T cells will secrete IL-2 which can then bind to the IL-2 high affinity receptor and activate the CD8 T cell.

Macrophage activation

-The enhancement of macrophage function -Requires interaction of peptide:MHC class II complexes on the macrophage with the TCR on a TH1 cell. -Causes phagosomes that contain captured microorganism to be more efficiently fused with lysosomes. -Increase synthesis by activated macrophages of highly reactiveandmicrobicidal molecules (oxygen radicals, nitric oxide (NO) and proteases) will kill engulfed pathogens.

The properties and functions of effector T cells

-The interaction of effector T cells with their specific antigen that are presented by APC or target cells. -After differentiating (secondary lymphoid organs) effector T cells detach from the APC that nursed their differentiation -Cytotoxic CD8 T cells, CD4 TH1, TH2, TH17 and Treg : ~Sites of infection +CD8 induces apoptosis in infected host cells +CD4 secrete cytokines -CD4 TFH: Stays in the secondary lymphoid organs ~CD4 secrete cytokines -T-cell effector functions are turned on when TCRs bind to peptide:MHC complexes This binding stimulates the T cell to release effector molecules that act on the target cell.(Tcell-->TCR-->Peptide:MHC--> Target cell)

Signals from TCR and co-receptors alter the pattern of gene transcription in activated T cells.

-The signal that antigen has bound the TCR is transmitted by the cytoplasmic tails of CD3 chains, associated with the TCR a- & B-chains. -CD3 cytoplasmic tails contain sequences called immunoreceptor tyrosine-based activation motifs (ITAMs),which associate with cytoplasmic protein tyrosine kinases. Kinases are activated by receptor clustering and phosphorylate ITAM tyrosine residues. Enzymes and other signaling moleculesbind to the phosphorylated tyrosine residues and become activated. Pathways of intracellular signaling are initiated to effect alterations in gene expression.

TFH cells stimulate the proliferation and differentiation of naïve B cells

-The specific interaction of an antigen binding Bcell with a TFH cell leads to the expressionof CD40 ligand (CD40L)on the T cell -Linked recognition - B cell may recognize a surface epitope but the peptide presented to the T cell may not be the same exact epitope (remember it will be a peptide). -For B cell to receive T cell help, both cells must recognize the same antigen - cognate interactions.

Activation of naïve T cells on encounter with antigen

-Upon infection, the immune system quickly recruits the small number of naïve T cells that are specific to the pathogen ---> make contact with antigens derived from the pathogen. -Recruitment involves secondary lymphoid tissues. Antigen is brought in from outlying tissues via lymph. Antigen encounters the T cells brought in via blood. 1.Examination of the activation of naïve T cells ---> effector T cells byprofessional APC (P-APC)within lymphoid tissues Dendritic cell 2.Interaction of a naïve T cell with antigen presented by cellsother than P-APCs leads to inactivation rather than activation of T cells -The immune system does not initiate the adaptive immune response wherever a pathogen creates a site of infection. -The immune system captures the pathogenand takes it to thesecondary lymphoid tissues. +Mediated by dendritic cells(migratory) +Process is same for infections in peripheral tissues, mucosal surfaces and blood

Cytokine Environment Plays Large Role in Type of T-cell Produced

-What type of effector T cell a CD4 T cell will become is dictated by cytokines in the immediate environment and dendritic cell activating it. -The local environment is created by the innate immune response at the site of infection and the dendritic cells, pathogens and cytokines that make their way to the secondary lymphoid tissue (ex; draining lymph node).

Antigen recognition without co-stimulation leads to a nonresponsive T cell

-When a TCR on a mature naïve T cell binds to a peptide:MHC complex on an APC that does not express the co-stimulatory molecule B7 the T cell becomes nonresponsive or anergic and cannot be activated with subsequent antigen encounter. -Some mature naïve T cells may be specific for self-proteinsexpressed by cells not found in the thymus. -These T cells will not be activated because the cells expressing these self antigens will not express the co-stimulatory molecule, B7. -A mature, naïve T cell that binds a self-peptide:MHC complex on a cell without B7 does not receive a co-stimulatory signaland undergoes anergy. +Anergic T-cells don't make IL-2---> no proliferation or differentiation -Anergic T cells can never be stimulated to proliferate or differentiate. -This process induces tolerance in the mature T cell repertoire.

Dendritic Cells Processes

-Wound in the skin -Langerhans' cells(immature dendritic cells) take up the antigen and migrate to a nearby lymph node. -Settle in the T-cell areas. -Differentiate into mature dendritic cells Skin and peripheral tissue infection = T cell response in draining lymph nodes. -Blood infections = antigen enter the spleen -Respiratory mucosa infection = tonsils or other bronchial-associated lymphoid tissue (BALT). -Gastrointestinal infections = Peyers patches, appendix or gut-associated lymphoid tissues (GALT) -Similar sequence of events in each case

Cytotoxins

-secreted cytotoxic proteins used to kill target cells. -Are specialized products of cytotoxic CD8 T cells.

T Cell-Mediated Immunity

1.Activation of naïve T cells on encounter with antigen What happens when naïve T cell encounters its specific antigen for the first time and is stimulated to differentiate into an effector cell? T-cell activation or T-cell priming 2.The properties and functions of effector T cellsDescribe the interaction of effector T cells with their specific antigen that are presented by APC or target cells.

Time course of programmed cell death

1.Contact is made with target cell and granules are distant from the point of contact. 2.After one minute, granules have begun to move toward the target cell. 3.After 3 minutes granules are almost ready to be released.

T-cell activation or T-cell priming 1st Stage

1.First stage of a primaryadaptiveimmune response 1. 6 kinds of effector T cells (you will learn about) -Cytotoxic CD 8 T cells (kill infected cells) -CD4 T cells- Function = secrete cytokines activate other immune cells -TH1 - Treg -TH2 - TH17 -TFH

Dendritic cells change their functions on taking antigen from infected sites to secondary lymphoid tissues

1.MHC class II stained green and a lysosomal protein is stained red. 2.Cell bodies are difficult to discern 3.Dendrites contain endocytic vesicles that stain both for MHC class II & lysosomal protein giving rise to yellow fluorescence (combination of red and green stain). 4.On activation and migration in the lymph to secondary lymphoid organs the morphology of the dendritic cell changes. 5.Dendritic cells stop phagocytosis indicated by a partial separation of MHC class II(green) from the lysosomal protein(red) 6.On reaching a lymph node"mature" dendritic cells begins antigen presentationto T-cells, stimulating the T cells instead of uptaking and processing antigen. 7.Lysosomal protein (red) is distinct from the MHC class II molecules (green). 8.MHC class II (green) is displayed at high density on the many dendritic processes.

Example of CD4 T cell increased sensitivity

Alternative splicing of RNA made from the CD45 genes causes activated T cells to express the CD45ROisoforms that associates with the TCR and CD4...this change makes the T cell more sensitive to stimulation by lower concentrations of peptide:MHC complexes.

Macrophages

Are found throughout the lymph node tissue and have several different functions: Are phagocytic cells that take up microbes and particles from the extracellular environment. -Degrade microorganism in phagolysosomes that are loaded with hydrolytic enzymes. -Trap and degrade pathogens--> this enables macrophages to process and present antigen to prevent infection from reaching blood. -Prevents noninfectious particulates from lymph nodes from entering blood and blocking small blood vessels. -Remove and degrade lymphocytes that die in secondary lymphoid tissues.

Dendritic cells

Are sentinels in all body tissues -Take up antigen, migrate to lymphoid organs, and present foreign antigens to naïve T cells -Activated with pathogen uptake and antigen processing and presentation by MHC I and II molecules. -Major function is the triggering T-cellresponses highly specialized and effective. Dendritic cells are migratory cells and have highest concentration of B7---> carry load of antigen from infection site---> nearest secondary lymphoid tissues.

Third Type of P-APC

B cell Ig binds specific antigen from extracellular environment. Ag:Ig complex---> internalized by receptor-mediated endocytosis ---> transported to endocytic vesicles---> degraded into peptides --->peptides bind MHC class II molecules --->peptide:MHC class II complex---> transported to cell surface In the primary immune response, the naïve B-cells are activated by effector T-cells that were activated in the same secondary lymphoid tissue.

The principal co-stimulatory molecules on professional APC are B7 molecules, which bind CD28 proteins on the T-cell surface

Binding of the T-cell receptor and its co-receptor CD4 to the peptide:MHC class II complex on the dendritic cell delivers a signal 1. This signal induces clonal expansion of T cells only when the co-stimulatory signal 2 is also given by the binding of CD28 (on the T cell ) to B7 (on the APC). Both CD28 and B7 are members of the immunoglobulin superfamily. There are two forms of B7, called B7.1 (CD80) and B7.2 (CD86), but their functional differences have yet to be understood. +Also known to as co-stimulatory molecules or co-stimulator molecules

Effector cytotoxic T cells contain lytic granules

CD8 T cells synthesize cytotoxins in inactive forms and package them into lytic granules when T cells are activated by specific antigen in the secondary lymphoid organs. CD8 T cells then migrate to sites of infection and will recognize specific peptides (made from proteins from infected cell) in the context of MHC class I molecules presented by the infected cell. TCR binding signals the cytotoxic T cell to secrete the contents of lytic granules directly onto a small localized areaon the surface of the infected target cell.

In absence of infection professional APC do not express co-stimulatory molecules

Capacity to activate naïve T cells is only acquired during an infection. B7 expressionis a direct consequence of infection,induced by interaction of a potential APC with microbial products via innate receptors (TLR's, mannose receptors...) that recognize pathogenic molecules (LPS, ds DNA, viral RNA...).

c-SMAC

Central supramolecular activation complex -Allows clustering of receptors-co-receptors and signaling molecules

Changes in adhesion molecule expression

Changes in the adhesion molecule expression of activated effector T cells versus resting naïve T cells also results in different patterns of migration. Without the expression of L-selectin effector T cells no longer recirculate through lymph nodes by leaving the blood through HEVs. Effector T cells instead express VLA-4 which enables bindingto endothelial cells of blood vessels in infected/inflamed tissues - where their effector function is needed. VLA-4 binds to the cell adhesion molecule VCAM-1 which is selectively expressed on the endotheliumof blood vessels in inflamed tissue.

Molecules that carry out T cell effector functions:

Cytokines Cytotoxins

any cytokine receptors signal through a pathway in which receptor-associated kinases activate transcript factors directly

Cytokines bind to cytokine receptors composed of two or three chains. The cytoplasmic tail of most cytokine receptors are associated with protein kinases called Janus kinases (JAKs). Cytokine binding causes dimerization of the cytokine receptors which stimulate the JAKs to phosphorylatemembers of a protein family called STATs (Signal Transducers and Activators of Transcription). On phosphorylation, two STAT molecules dimerize and move from the cytoplasm to the nucleus where they activate specific genes. The genes that are activated is determined by the cytokine/sthat were bound.

Immature DC

DEC205 facilitates receptor mediated endocytosis and pinocytosis of antigens. -ex: langerhans cells

Relaxed activation requirementsfor effector T cells

Effector T cells can respond to their specific antigenwithout the need for co-stimulation via B7-CD28 interaction. Effector T cells can respond to antigen presented on cells other than professional antigen-presenting cells. Benefits ex: Cytotoxic CD8 Teffector cell can bind to MHC I + pathogen peptide on epithelial cell and tell the epithelial cell to die by apoptosis.

Microbial substances induce co-stimulatory activity in macrophages

Endocytosis of bacteria by dendritic cells and their breakdown in phagolysosomes leads to the release of substances such as bacterial lipopolysaccharide which induce the expression of Co-stimulatory B7 molecules. Peptides derived from the degradation of bacterial protein in the dendritic cells vesicular system are bound by MHC class II molecules---> presented on the DC's surface. Activation of naïve T cells--->CD28 binding to B7 along with peptide:MHC complexes binding to TCR and co-receptor binding.

Activated effector T cell VERSUS Resting naïve T cells

How are they different? 1.Types of cell surface molecules expressed 2.Abundance of these cell surface molecules

Dendritic cells are developmentally related to macrophages

Immature dendritic cells use receptor mediated endocytosis to bring in microbial antigens using DEC 205 receptors(among others) -Antigens can also be taken up nonspecifically by macropinocytosis in which a cell engulfs extracellular fluid (ECF). Important for initiation of T-cell viral responses -Dendritic cells acquire viral antigens via infection or by taking up virus particles from the ECF or other infected cells .

Studying Ab and T-cell responses in lab animals

Immunization of protein antigen alonerarely induces an immune response. Protein antigens + certain bacterial breakdown products (adjuvants) are required for a strong immune response. Microbial products, known as adjuvants, induce co-stimulatory activity in P-APC's. -Whole microorganism = more effective vaccines than highly purified Ag molecules -Mechanism which allows the immune system to distinguish between antigens borne by infections agentand antigens associated with innocuous proteins.

Mature DC

Increase expression of B7 co-stimulators, MHC moleculesand adhesions molecules like DC-SIGN -ex: interdigitating reticular cells

TH1

Increases inflammation at the site of infection -Main defense against intracellular viral and bacterial infections -Activated by IL-12 (dendritic cells and macrophages) and IFN-y secreted by NK cells during innate immune response -Activation of TF T-bet ---> differentiation into TH1 effector

Activation of naïve T cells requires a co-stimulatory signal delivered by a professional antigen-presenting cell.

Initial intracellular signal generated by TCR:peptide:MHC complex = necessary for naïve T cell activation but... Not sufficient.....a second co-stimulatory signal is required. Co-stimulatory signals delivered by APC. Both the antigen-specific stimulation and the co-stimulation must be delivered by ligands on the same APC.

Dendritic cells are developmentally related to macrophages Cont.

Langerhans' cell of the skin is a typical immature dendritic cell On skin infection, local Langerhans' cells will take up and process microbial antigen before traveling to the T-cell cortex areas of the draining lymph node and maturing to become a professional APC. In lymph nodes, mature dendritic cells have a distinctive morphology which led to them being called interdigitating reticular cells. Activated dendritic cells not only express B7 and class II MHC molecules but also express high levels of adhesion molecules (i.e. DC-SIGN) and secrete the chemokine CCL18 specifically attracts naïve T-cells. +DC-SIGN binds to ICAM-3 on the T-cell

TH1 CD4 cells induce macrophages to become activated

Macrophage have receptors that bind to microorganisms and facilitate their phagocytosis, destruction and intracellular degradation. As a result, macrophages present pathogen-derived peptide on MHC class II molecules toactivateeffector TH1 cells. Some microorganisms can adapt to the macrophage and interfere with macrophage function by living and replicatinginside the phagosome. Thus, a principal function of TH1 cells is to activate macrophages to: -Increase their phagocytic ability. -Increase their capacity to kill ingested microorganisms.

TH1 CD4 cells activate macrophages to become highly microbicidal

Macrophages require two signals for activation, both of which are delivered by TH1 cells. -The primary signal is provided by IFN- y, the characteristic cytokine produced by TH1 cells. -The second signal is delivered by the CD40 ligand(CD40L) on T cells interacting with the CD40 receptor on macrophages. When a TH1 cell specific for a bacterial peptide contacts a macrophage that presents that peptide, the TH1 cell is induced to secrete IFN- y and express CD40 ligand. Together these newly synthesized proteins activate the macrophage to kill the bacterial living inside of its vesicles. •CD8 T cells can also produce IFN- y and can aid in the activation of macrophages. •However, microbicidal substances produced by macrophages can be harmful to tissues. •CD4 TH2 cells produce cytokines that inhibit macrophage activation, showing the TH2 cells can control the TH1 response.

Dendritic cells develpoment

Movement of dendritic cell from site of infection in the periphery to a secondary lymphoid organ causes changes in the dendritic cell's surface molecules, functions and morphology. In tissues, dendritic are active in the capture, uptake and processing of antigen - called immature dendritic cells. In secondary lymphoid tissues, dendritic cells gain the capacity to interact with T cells - called mature or activated dendritic cells (no longer phagocytic). Mature dendritic cells have finger-like processes called dendrites that contact T cells in the cortex of lymph node.

Binding of L-selectin to vascular addressins directs naïve lymphocytes homing to lymphoid tissues.

Movement of naïve T cells into secondary lymphoid tissues Homing determined by T-cell surface selectin (L-selectin) interacting with two vascular addressins on surface of HEV venules (CD34 & GlyCAM-1) L-selectin on naïve T cells (and naïve B cells) binds to sulfated carbohydrate sialyl-Lewisxmoieties of vascular addressins CD34 and GlyCAM-1 on the high endothelial cells of lymph venules

Recirculating, Naïve T cells encounter antigen during recirculation through lymphoid organs

Naïve T cells (blue & green) recirculate through secondary lymphoid organs, such as the lymph node. These leave the blood through HEVs and enter the lymph node cortex, where they mingle with mature dendritic cells. T cells that do not encounter their specific cells and antigen(green) leave the lymph node in the efferent lymph and eventually rejoin the bloodstream. T cells that encounter antigen (blue) on antigen-presenting cells are activated to proliferate and to differentiate into effector cells. These effector T cells can also leave the lymph node in the efferent lymph and enter the circulation

Cytotoxic CD8 T cells are selective and serial killers of target cells at sites of infection

Once inside of a cell, the pathogen becomes inaccessible to antibody and other immune system proteins (i.e. complement). Elimination will be either through the efforts of the infected cell itself or by direct attack on the infected cell by the immune system. Function of cytotoxic CD8 T cells is to kill cells that have become overwhelmed by intracellular infection. -Infected cell is sacrificed to prevent spread of infection to healthy cells.

Proliferation and differentiation of activated T cells are driven by the cytokine interleukin-2 (IL-2)

One of the most important genes to be transcribed codes for the cytokine IL-2 -Activation of T cell by professional APC initiates a program of differentiation controlled by the cytokine IL-2. A burst cell division Acquisition of effector function -IL-2 is synthesized and secreted by activated T cells. -IL-2 binds IL-2 receptors of T cell to drive clonal expansion of the activated cell -IL-2 is one of a number of cytokines produced by activated and effector T cells to control the development differentiation of cells in the immune response. -IL-2 production requires both the signal delivered through the TCR:co-receptor complex and the co-stimulatory signal delivered through CD28. Signals through the TCR:CD3 complex activate NFAT activate transcription of the IL-2 gene (IL-2 mRNA is inherently unstable). Co-stimulation (CD28:B7) stabilizes the IL-2 mRNA which causes an increase in the synthesis of IL-2 by T cells (20-30X). Co-stimulation also activates other transcription factorsthat stimulate the transcription of the IL-2 gene (3X). Principal effect of co-stimulation is to increase synthesis of IL-2 by 100 fold.

In addition to cytotoxic functions, CD8 cells also contribute to the immune response by secreting cytokines

Secretion of interferon-gamma (IFN-)inhibits the replication of viruses in the infected cells IFN-y also increases the processing and presentation of viral antigen by MHC class I molecules. IFN-yactivates macrophages in the vicinity of the cytotoxic Tcells. Activated macrophages then get rid of dying infected cells helping damaged tissue to repair.

Cytotoxic CD8 T cells kill infected cells selectively

Specific recognition of peptide:MHC complex on an infected cell by a cytotoxic CD8 T cell (CTL) programs the infected cell to die and leaves healthy cells alone. The CTL detaches from the first target cell (which dies), synthesizes a new set of lytic granules and then seeks out and kills another infected cell (new target).

Cytotoxic T cells kill their targets by inducing apoptosis - two pathways

T cells induce apoptosis by two pathways (1) initiated by cytotoxins and (2) interactions with cell surface molecules: Combination of perforin, granulysin and serglycin form pores in target cells membrane. Also may use granzymes.

Professional antigen-presenting cells versus other antigen-presenting cells

The characteristic that distinguishes professionalantigen-presenting cells (APCs) from other antigen-presenting cells is the presence of B7 co-stimulatory molecules on their surface. The three kind of professional APCs are: the dendritic cell, the macrophage and the B cell. The one cell we know can activate naïve T-cells is the Dendritic cell.

Leukocyte adhesion molecules

The four structural classes of adhesion molecule present on white blood cells and the cells with which they interact are: 1.Selectins - are carbohydrate-binding lectins 2.Vascular addressins - contain carbohydrate groups to which selectins bind 3.Integrins 4.Proteins in the immunoglobulin superfamily

On activation, CD4 T cells can acquire different helper functions

Towards the end of the proliferative phase, T cells acquire the capacity to synthesize the proteins they need to perform their specialized functions. CD4 T (helper) cells produce cell-surface molecules and soluble cytokines to activate and help other cell types.-Macrophages, B cells, neutrophils and suppress effector T cells CD4 T cells differentiate into CD4 TH1, TH2, TH17, TFH and Treg cells.

Transient adhesive interactions between T cells & dendritic cells are stabilized by specific Ag recognition

When a naïve T cell binds to its specific ligand on an antigen-presenting dendritic cell, intracellular signaling through the TCR induces a conformational change in LFA-1 that causes it to bind with higher affinity to ICAMs on the antigen-presenting cell. These T-cells will suppress expression of S1P receptors. This interaction is stable and can last several days during which the T cell proliferates and its progeny differentiates into effector cells. If antigen isn't met, then the T-cells will be drawn out of the lymph node by a different chemotactic molecule called sphingosine 1-phosphate (S1P). +Now the T-cell can continue to recircluate until it meets it's antigen.

Paracrines

act locally on another cell

Autocrines

act on the cell that produced them

Granzymes

activate apoptosis by activating caspases in host cell---> cascade ending in apoptosis

Lytic granules

are modified lysosomes with a mixture of cytotoxins.

Cytokines produced by TH1 cells

can reinforce the differentiation of TH1 cells (cell mediated response) and Cytokines produced by TH2 cells can reinforce the differentiation of TH2cells (humoral response) leading to a positive feedback loop = polarized

Necrosis

death due physical or chemical injury in which cells lyse and disintegrate.

Perforin

directs other cytotoxins into cytoplasm of infected host cell

Granulysin

has antimicrobial activity and can activate apoptosis

Apoptotic cell in latestages

has very condensed nucleus, no mitochondria, and the cytoplasmand cell membranes are largely lost through vesicle shedding.

p-SMAC

peripheral supramolecular activation complex -Adhesion molecules - forms tight seal

Serglycin

scaffold for other cytotoxins to be carried on

Apoptotic cell in early stages

showing chromatin condensation (red) but well-defined plasma membrane that is shedding vesicles.