IMMUNE RESPONSE

The Immune Response

Immunity: Free from burden. Ability of an organism to recognize and defend itself against specific pathogens or antigens. Immune Response: Third line of defense. Involves production of antibodies and generation of specialized lymphocytes against specific antigens. Antigen: Molecules from a pathogen or foreign organism that provoke a specific immune response.

Immune Response
Specific immunity induced in a host by an antigenic stimulus. 2 types: Humoral (Antibody mediated) immunity Cellular (cell mediated) immunity Both develop together, one or other may predominate.

The Immune System is the Third Line of Defense Against Infection

I. Humoral (Antibody-Mediated) Immunity

Involves production of antibodies against foreign antigens. Antibodies are produced by a subset of lymphocytes called B cells. B cells that are stimulated will actively secrete antibodies and are called plasma cells. Antibodies are found in extracellular fluids (blood plasma, lymph, mucus, etc.) and the surface of B cells. Defense against bacteria, bacterial toxins, and viruses that circulate freely in body fluids, before they enter cells. Also cause certain reactions against transplanted tissue.

Humoral Immune Response

Production of antibodies consists of 3 steps: Entry of antigen, its distribution and fate in the tissues and its contact with appropriate immunocompetent cells. Processing of Ag by cells and control of Ab forming process (central functions) Secretion of Ab its distribution in tissues and body fluids and mainfestation of its effect.

 Primary responses: Ab response to an initial Ag slow, sluggish, shortlived.

Ab IgM Ab more specific, less avid called early Ab

Secondary responses: response to subsequent stimuli with the same Ag prompt, powerful and prolonged. Ab IgG Late Ab

Fate of Ag in tissues
Fate of Ag depends on factors like physical and chemical nature of Ag, its dose, route of entry and if stimulus is primary or secondary. Ags introduced IV are localised in liver, kidney, BM etc and broken by RE cells and excreted in urine. Ags introduced subcutaneously are mainly localised in draining lymphocytes. Particulate Ags may be removed in 2 phases: 1. Nonimmune phase Ag is engulfed by phagocytic cells, broken down and eliminated with appearance of specific Ab. 2. Phase of immune elimination - Ag-Ab complexes are formed and Ags phagocytosed. Soluble Ags show 3 phases during elimination: Equilibration- Ag diffuse in to extravascular spaces. Metabolism level of Ag falls due to catabolic decay Immune elimination elimination of Ag due to Ag-Ab complex formation.

 Ag presentation to immunocompetent cells occurs in 2 ways by macrophages and dendritic cells in lymphnode follicles. Ag processing by macrophages appear to be essential for the primary IR to many Ags, but not for secondary IR. Ag capture of the dendritic cells of the lymphnode follicles occur in the presence of pre-existing Ag. Both dendritic cells and macrophages present Ag in areas where there is ample lymphocytes so that the appropriate lymphocytes can recognise the Ag and initiate IR.

CELLULAR IMMUNE RESPONSE

CMI refers to specific IR that do not involve Abs. The only facet of CMI that was investigated to any extent was delayed hypersensitivity. CMI is more important than AMI. CMI depends on the actions of Tlymphocytes.

 T lymphocytes are divided into different sub populations based on functional capacity. Regulator T cells : a) T -helper cells (TH) b)T- suppresor cells (TS) Effector T cells: a) cytotoxic T-lymphocytes (CTL) b) Delayed type hypersensitivity (DTH) c) Mixed lymphocyte reactivity (MLR)

 TH cells facilitate B cell response to many Ags. Ts inhibit Ab production by B-cells, and immune reactions by effector T-cells. IR regulated by balanced activity of TH and TS cells. Over activity of TH or decreased acitvity of TS cause abnormal IR Autoimmunity. Diminished TH function or increased TS cell activity Immunedeficiency

 CMI funcitons: Delayed hypersensitivity Immunity to infectious diseases caused by obligate and facultative intracellular parasites. Eg infections with bacteria (TB, leprosy) Fungi, protozoa, viruses (small pox, measles, mumps) Transplantation immunity and graft Vs host rxns Immunological surveillance and immunity against cancer Pathogenesis of certain autoimmunediseases (thyroiditis, encephalomyelitis)

 Induction of CMI sensitise T-lymphocytes against Ag. When the T-lymphocyte contacts the Ag determinant - blast transformation and clonal proliferation selectively in paracortical areas of the lymphnodes. Activated lymphocytes release biologically active products lymphokines responsible for CMI. Macrophages and other macronuclear cells under the effect of lymphokine effect the destruction of M.O and other processes in CMI.

Antibodies are Produced by B Lymphocytes

Duality of Immune System (Continued) II. Cell Mediated Immunity

Involves specialized set of lymphocytes called T cells that recognize foreign antigens on the surface of cells, organisms, or tissues:
Helper T cells Cytotoxic T cells

T cells regulate proliferation and activity of other cells of the immune system: B cells, macrophages, neutrophils, etc. Defense against:
Bacteria and viruses that are inside host cells and are inaccessible to antibodies. Fungi, protozoa, and helminths Cancer cells Transplanted tissue

Cell Mediated Immunity is Carried Out by T Lymphocytes

Antigens
Most are proteins or large polysaccharides from a foreign organism.
Microbes: Capsules, cell walls, toxins, viral capsids, flagella, etc. Nonmicrobes: Pollen, egg white , red blood cell surface molecules, serum proteins, and surface molecules from transplanted tissue.

Lipids and nucleic acids are only antigenic when combined with proteins or polysaccharides.

Molecular weight of 10,000 or higher.

Hapten: Small foreign molecule that is not antigenic. Must be coupled to a carrier molecule to be antigenic. Once antibodies are formed they will recognize hapten.

Antigens
Epitope: Small part of an antigen that interacts with an antibody. Any given antigen may have several epitopes. Each epitope is recognized by a different antibody.

Epitopes: Antigen Regions that Interact with Antibodies

Antibody Structure

How Do B Cells Produce Antibodies?

B cells develop from stem cells in the bone marrow of adults (liver of fetuses). After maturation B cells migrate to lymphoid organs (lymph node or spleen). Clonal Selection: When a B cell encounters an antigen it recognizes, it is stimulated and divides into many clones called plasma cells, which actively secrete antibodies.

Each B cell produces antibodies that will recognize only one antigenic determinant.

Clonal Selection of B Cells is Caused by Antigenic Stimulation

Humoral Immunity (Continued) Apoptosis

Programmed cell death (Falling away). Human body makes 100 million lymphocytes every day. If an equivalent number doesnt die, will develop leukemia. B cells that do not encounter stimulating antigen will self-destruct and send signals to phagocytes to dispose of their remains. Many virus infected cells will undergo apoptosis, to help prevent spread of the infection.

Humoral Immunity (Continued) Clonal Selection

Clonal Selection: B cells (and T cells) that encounter stimulating antigen will proliferate into a large group of cells. Why dont we produce antibodies against our own antigens? We have developed tolerance to them. Clonal Deletion: B and T cells that react against self antigens appear to be destroyed during fetal development. Process is poorly understood.

Consequences of Antigen-Antibody Binding

Antigen-Antibody Complex: Formed when an antibody binds to an antigen it recognizes. Affinity: A measure of binding strength. 1. Agglutination: Antibodies cause antigens (microbes) to clump together.
IgM (decavalent) is more effective that IgG (bivalent). Hemagglutination: Agglutination of red blood cells. Used to determine ABO blood types and to detect influenza and measles viruses.

2. Opsonization: Antigen (microbe) is covered with antibodies that enhances its ingestion and lysis by phagocytic cells.

Consequences of Antibody Binding

Humoral Immunity (Continued)

3. Neutralization: IgG inactivates viruses by binding to their surface and neutralize toxins by blocking their active sites. 4. Antibody-dependent cell-mediated cytotoxicity: Used to destroy large organisms (e.g.: worms). Target organism is coated with antibodies and bombarded with chemicals from nonspecific immune cells. 5. Complement Activation: Both IgG and IgM trigger the complement system which results in cell lysis and inflammation.

Consequences of Antibody Binding

Immunological Memory
Antibody Titer: The amount of antibody in the serum. Pattern of Antibody Levels During Infection Primary Response: After initial exposure to antigen, no antibodies are found in serum for several days. A gradual increase in titer, first of IgM and then of IgG is observed. Most B cells become plasma cells, but some B cells become long living memory cells. Gradual decline of antibodies follows.

Immunological Memory secondary Response:

Subsequent exposure to the same antigen displays a faster and more intense antibody response. Increased antibody response is due to the existence of memory cells, which rapidly produce plasma cells upon antigen stimulation.

Antibody Response After Exposure to Antigen

T Cells and Cell Mediated Immunity

Antigens that stimulate this response are mainly intracellular. Requires constant presence of antigen to remain effective. Unlike humoral immunity, cell mediated immunity is not transferred to the fetus. Cytokines: Chemical messengers of immune cells. Over 100 have been identified. Stimulate and/or regulate immune responses.
Interleukins: Communication between WBCs. Interferons: Protect against viral infections. Chemokines: Attract WBCs to infected areas.

T Cells and Cell Mediated Immunity

Cellular Components of Immunity: T cells are key cellular component of immunity. T cells have an antigen receptor that recognizes and reacts to a specific antigen (T cell receptor). T cell receptor only recognize antigens combined with major histocompatability (MHC) proteins on the surface of cells.
MHC Class I: Found on all cells. MHC Class II: Found on phagocytes.

Clonal selection increases number of T cells.

T Cells Only Recognize Antigen Associated with MHC Molecules on Cell Surfaces

T Cells and Cell Mediated Immunity Types of T cells

1. T Helper (TH) Cells: Central role in immune response.
Most are CD4+ Recognize antigen on the surface of antigen presenting cells (e.g.: macrophage). Activate macrophages Induce formation of cytotoxic T cells Stimulate B cells to produce antibodies.

Central Role of Helper T Cells

Types of T cells
2. Cytotoxic T (Tc) Cells: Destroy target cells.
Most are CD4 negative (CD4 -). Recognize antigens on the surface of all cells:
Kill host cells that are infected with viruses or bacteria. Recognize and kill cancer cells. Recognize and destroy transplanted tissue.

Release protein called perforin which forms a pore in target cell, causing lysis of infected cells. Undergo apoptosis when stimulating antigen is gone.

Cytotoxic T Cells Lyse Infected Cells

Types of T cells
3. Delayed Hypersensitivity T (TD) Cells: Mostly T helper and a few cytotoxic T cells that are involved in some allergic reactions (poison ivy) and rejection of transplanted tissue. 4. T Suppressor (Ts) Cells: May shut down immune response.

Nonspecific Cellular Components

1. Activated Macrophages: Stimulated phagocytes.
Stimulated by ingestion of antigen Larger and more effective phagocytes. Enhanced ability to eliminate intracellular bacteria, virus-infected and cancerous cells.

2. Natural Killer (NK) Cells:

Lymphocytes that destroy virus infected and tumor cells. Not specific. Dont require antigen stimulation. Not phagocytic, but must contact cell in order to lyse it.

Relationship Between Cell-Mediated and Humoral Immunity 1. Antibody Production

T-Dependent Antigens:
Antibody production requires assistance from T helper cells. A macrophage cells ingest antigen and presents it to TH cell. TH cell stimulates B cells specific for antigen to become plasma cells. Antigens are mainly proteins on viruses, bacteria, foreign red blood cells, and hapten-carrier molecules.

T-Independent Antigens:
Antibody production does not require assistance from T cells. Antigens are mainly polysaccharides or lipopolysaccharides with repeating subunits (bacterial capsules). Weaker immune response than for T-dependent antigens.

Humoral Response to T Dependent Antigens

Humoral Response to T Dependent Antigens

Relationship Between Cell-Mediated and Humoral Immunity 2. Antibody Dependent Cell Mediated Cytotoxicity
Target cell is covered with antibodies, leaving Fc portion sticking outwards. Natural killer and other nonspecific cells that have receptors for Fc region are stimulated to kill targeted cells. Target organism is lysed by substances secreted by attacking cells. Used to destroy large organisms that cannot be phagocytosed.

Destruction of Large Parasites by Antibody Dependent Cell-mediated CytotoxicityADCC

Overview of the Immune Response

Major Histocompatibility Complex

MHC cluster of genes in mammals. Associated with intercellular recognition and with self and non-self recognition. A region of multiple loci that plays a imp role in determining if the transplanted tissue will be accepted as self (histocompatible)/ rejected (histoincompatible) MHC collection of genes on chr 6 in humans and chr 17 in mice. MHC in humans Human Leukocyte Antigen (HLA), in mice H-2 complex Present in almost all tissues on PM

 3 classes of MHC: Class I MHC found on almost all types of nucleated body cells. Class II MHC found on leukocytes involved in T helper cell related IR (macrophages, APCs, B cells) Class III MHC include various secreted proteins with immune functions. (complement component c2, c4 factor B), inflammatory cytokines, tumor necrosis factors. These are not membrane protiens, hence no role in antigen presentation.

 Class I MHC : Complex of 2 protein chains one with mass 45 Kda (heavy chain), other with mass 125 Kda (2 microblobulin) 2 chains have 4 regions Outer most segment of heavy chain divided in to 3 functional domains 1,2,3 2 M and 3 are non covalently associated with one another and are close to PM. 1,2 domain remain outside and form Ag binding pocket.

 Class II MHC: Transmembrane protein with & chains of mass 34 and 28 kda Both folded to give 2 domains.

 The presence of foreign peptides in MHC groove alerts immune system and activates Tcells which in turn activate macrophages. Class I and II molecules present peptides that arise in different places within cells as a result of Antigen processing. Class I molecule binds to peptides that originate in cytoplasm.

 Endogenous Ag protiens are digested inside cell as a part of natural process by which cells renew its protiens. The short peptide fragments that result are pumped by a specific transporter protien from cytoplasm into ER. In ER the cl I MHC chain is synthesised-associates with 2 microglobulin. This dimer binds to peptide on entering ER. The MHC and peptide are then carried and anchored to PM If peptide is foreign- eg short pieces of viral protein- a passing CD8+ T cell (cytotoxic T-lymphocytes) whose T-cell receptor is specific for antigenic peptide will bind to peptide MHC complex and kill the infected cell.

 Cl II MHC bind to fragments from exogenous Ag like bacteria and virus. The APC takes in the Ag by receptor mediated endocytosis/phagocytosis, and produce Ag fragments by digestion in phagolysosome. Fragments combine with preformed class II MHC and delivered to cell surface. Peptide is recognised by CD4+ TH cells. These cells do not directly kill the target, instead 2 responses: 1.To enlarge and divide thereby increase no.of CD4 cells that can react to the Ag 2. to secrete cytokines (IL 2) that inhibit pathogen that produce Ag.