Slide 1. Slide 2. Slide 3 IMMUNOLOGY AND THE PATHOPHYSIOLOGY OF INFECTION

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1 Slide 1 IMMUNOLOGY AND THE PATHOPHYSIOLOGY OF INFECTION Pharmacotherapy of Infectious Diseases 5214 Slide 2 IMMUNE SYSTEM A network of cells, tissues and organs that work together to protect the body against foreign invaders, including substances, cells and tissues, through a constellation of responses The immune response is specific The immune response has memory Slide 3 BASICS Why? How? Elements of the Immune System and Their Roles in Defense Origin of immune system components Antibody Structure Organs of the immune system

2 Slide 4 BASICS Why? How? Elements of the Immune System and Their Roles in Defense Origin of immune system components Antibody Structure Organs of the immune system Slide 5 SURFACE BARRIERS Slide 6 INFLAMMATION

3 Slide 7 RECOGNITION AND DESTRUCTION Slide 8 DEFINITIONS Adjuvant: substance that promotes immune response to immunogen Antigen: substance recognized by immune response Carrier: protein modified by hapten to elicit response Epitope: minimal molecular structure recognized by immune response Hapten: incomplete immunogen that cannot initiate response but can be recognized by antibody Immunogen: substance capable of eliciting an immune response Slide 9 BASICS Why? How? Elements of the Immune System and Their Roles in Defense Origin of immune system components Antibody Structure Organs of the immune system

4 Slide 10 HEMATOPOIESIS Slide 11 CELLS OF THE IMMUNE SYSTEM Three Types of Blood Cells 1. Erythrocytes red blood cells 2. Thrombocytes - platelets 3. Leukocytes white blood cells (WBC) Slide 12 NORMAL ADULT COMPLETE BLOOD COUNT (CBC W/ DIFFERENTIAL) Cell Type Mean Number per L Percentage Red Blood Cells Platelets WBC (leukocytes) Neutrophils Lymphocytes Monocytes Eosinophils Basophils ,000 7,400 4,400 2, % 50 70% 20 40% 1 6% 1 3% < 1%

5 Slide 13 ROLE/FUNCTION Slide 14 FUNCTIONAL PROPERTIES OF CELLS Coordinate immune response by communicating with other cells Recognition Destruction Slide 15 COMMUNICATION

6 Slide 16 IMMUNE SYSTEM COMMUNICATION Direct cell to cell contact Cytokines Small secreted proteins that mediate and regulate the immune response Interferons (IFN) Produced in response to viral infections (INF-, INF - ) or upon activation of the immune response (INF- ) Lymphokines Cytokines made by lymphocytes Slide 17 SPECIFIC TYPES OF CYTOKINES Chemokines Cytokines with chemotactic activities Attracts leukocytes to infection sites Associated with inflammatory response Interleukins (IL) Cytokines made by one leukocyte which act on other leukocytes Slide 18 FUNCTIONAL PROPERTIES OF CELLS Coordinate immune response by communicating with other cells Recognition Destruction

7 Slide 19 FUNCTIONAL PROPERTIES OF CELLS Antigen Presenting Cells (APC) A cell that displays proteins of a foreign cell on its surface (epitope) to alarm and activate other immune cells Cell Destruction Phagocytosis Attachment Internalization digestion Release lymphotoxins which cause cell lysis Slide 20 CELLS OF THE IMMUNE SYSTEM Slide 21 CELLS OF THE IMMUNE SYSTEM

8 Slide 22 CELLS OF THE IMMUNE SYSTEM Cells NK cells Phagocytic Cells Neutrophils Eosinophils Monocytes* Immature dendritic cells Dendritic cells* Langerhans cells* Macrophages* Characteristics and Functions Large, granular lymphocytes Markers: Fc receptors for antibody, KIR Kill antibody-decorated cells and virus-infected or tumor cell (no MHC restriction) Granulocytes with short life span, multilobed nucleus and granules, segmented band forms (more immature) Phagocytose and kill bacteria (polymorphonuclear leukocytes) Bilobed nucleus, heavily granulated cytoplasm, stains with eosin Involved in parasite defense and allergic response Horseshoe-shaped nucleus, lysosomes, granules Precursors to macrophage-lineage and dendritic cells, cytokine release Blood and tissue Cytokine response to infection, process antigen Lymph nodes, tissue Most potent APC; initiates and determines nature of T-cell response Present in skin Same as immature dendritic cell Possible residence in tissue, spleen, lymph nodes, and other organs; activated by IFN-γ and TNF Markers: large, granular cells; Fc and C3b receptors (M2) Remove debris, maintain normal tissue function, and facilitate repair, APC (M1) Activated cells initiate inflammatory and acute-phase response; activated cells are antibacterial, APC Cells Characteristics and Functions Slide 23 Antigen-Responsive T Cells T cells (all) α/β TCR CD4 T cells α/β TCR CD8 T-killer cells NKT cells Antibody-Producing Cells B cells Plasma cells Other Cells Basophils/mast cells Platelets Mature in thymus; large nucleus, small cytoplasm Markers: CD2, CD3, T-cell receptor (TCR) Helper/DTH cells; activation by APCs through class II MHC antigen presentation Produce cytokines; stimulate T- and B-cell growth; promote B-cell differentiation (class switching, antibody production) TH1 subtype (IL-2, IFN-γ, LT production): promote antibody and cell-mediated defenses (local and systemic), including DTH, CD8 T killer cells TH2 subtype (IL-4, IL-5, IL-6, IL-10 production): promote humoral (antibody) responses (systemic) TH17 subtype (IL-17, TNF-α, IL-6): stimulate epithelial cells and neutrophils and inflammation T regulator (Treg) cells (TGF-β, IL-10): control CD4 and CD8 T-cell activation and other cells; important for immunotolerance Recognition of antigen presented by class I MHC antigens Kill viral, tumor, nonself (transplant) cells; secrete cytokines Express NK cell receptors, TCR, and CD3 Rapid response to infection, cytokine release Mature in bone marrow (bursal equivalent), Peyer patches Large nucleus, small cytoplasm; activation by antigens and T-cell factors Markers: surface antibody, class II MHC antigens Produce antibody and present antigen Small nucleus, large cytoplasm Terminally differentiated, antibody factories Granulocytic Marker: Fc receptors for IgE Release histamine, provide allergic response, are antiparasitic Release clotting factors, antimicrobial peptides, chemokines and cytokines upon activation Slide 24 Neutrophils Eosinophils Basophils Monocytes Dendritic cells Macrophages Lymphocytes B and T cells Natural Killer Cells LEUKOCYTES

9 Slide 25 CELLS OF THE IMMUNE SYSTEM Neutrophil (50 70% WBC) A phagocyte also known as a polymorphonuclear leukocyte or seg Serves as primary human defense against bacteria Migrates from the blood to the infection site in response to chemotactic factors Recognizes and binds to immunoglobulin and complement coated via receptors on its surface Slide 26 CELLS OF THE IMMUNE SYSTEM Neutrophil left shift During an infection, the neutrophil count increases in blood Includes the presence of precursors or immature forms called bands (non-segmented nucleus) Slide 27 CELLS OF THE IMMUNE SYSTEM Eosinophil (1-3% WBC) A phagocyte Releases toxic contents of granules to burn holes in cells Primary role is to kill parasites; minor role against bacteria Accumulate in the face of an allergic reaction Basophil (< 1% WBC) Not a phagocyte Contain heparin and large amount of histamine Play a role in Type I hypersensitivity (allergic) reaction

10 Slide 28 CELLS OF THE IMMUNE SYSTEM Monocytes Phagocytes that circulate in the blood Develop into Macrophages Dendritic Cells Slide 29 MACROPHAGE Functions Phagocytosis Antigen presentation to T-cells Bind to immunoglobulin via cell surface receptor for IgG (facilitates phagocytosis) Bind to complement facilitates phagocytosis) Secrete cytokines Are activated by T cells & cytokines Slide 30 Functions Phagocytosis Most efficient antigen presenters Professional APC Determines nature of T-Cell response Secrete cytokines DENDRITIC CELL

11 Slide 31 CELLS OF THE IMMUNE SYSTEM Lymphocytes Natural Killer Cells Circulate in the blood and lymph Contain no specific surface markers Lyse (induce rupture) cancer cells and virus infected cells B cell name based on maturation in the bone marrow T cell name based on maturation in the thymus Slide 32 B AND T CELLS B cells Surfaces also contain immune globulin Main role Induce antibody production Ag presentation to T-cells (APC) T cells Surfaces also contain receptors (TCR) that recognize antigen on the surface of other cells Serve as the basis of cellmediated immunity Slide 33 TYPES OF T CELLS CD8 + T cells (Cytotoxic Cells) Recognize Ag presented by cells Kill cells expressing appropriate Ag Release lymphotoxins which cause cell lysis Also recognize & kill virally infected cells, foreign tissue (non - self) and tumor cells Inhibit the production of cytotoxic T cells once they are no longer needed Down regulate the activity of other cells CD4 + T cells (Helper Cells) Coordinate immune response by communicating with other cells Secrete cytokines which stimulate cytotoxic T cells and B cells to grow and differentiate Activate macrophages

12 Slide 34 MEMORY CELLS Memory T cells Programmed to recognize and respond to a antigen once it has invaded and been repelled Memory B cells Programmed to recognize and produce antibody to a antigen once it has invaded and been repelled Slide 35 SUMMARY IMMUNE SYSTEM COMPONENT ROLES Macrophage Dendritic cell Neutrophil T cell B cell Cytokines Slide 36

13 Slide 37 BASICS Why? How? Elements of the Immune System and Their Roles in Defense Origin of immune system components Antibody Structure Organs of the immune system Slide 38 IMMUNOGLOBULINS (IG) A glycoprotein produced in response to an antigen that is specific for the antigen and binds to it A large family of proteins also known as antibodies that serve a major role in humoral immunity Produced by plasma cells which evolve from B cells May act as Antigen receptors on the surface of B cells Antibodies that inactivate antigens Slide 39 IMMUNOGLOBULIN ISOTYPES

14 Slide 40 IMMUNOGLOBULIN STRUCTURE 4 polypeptide chains 2 light chains 2 heavy chains Each chain has Variable region Specific for antigens Recognize and bind antigen Constant region Consistent among isotype Slide 41 IMMUNOGLOBULIN ISOTYPES IgG (85%) Four subclasses exist that differ in structure Activates complement Binds to phagocytic cell surfaces Transferred via the placenta IgM (5-10%) First Ab produced in response to an antigenic challenge Exists as a pentamic molecule Serves as a receptor on B cell surface Most efficient Ab for activating complement Slide 42 IMMUNOGLOBULIN ISOTYPES IgA (5-15%) Two subclasses exist May exist as a monomer, dimer, trimer and multimer Concentrates in body fluids to guard against entrances to the body IgE (<1%) Protects against parasites Binds to mast cell surfaces and elicits allergic response Responsible for anaphylactic hypersensitivity reactions IgD (<1%) Acts as a B cell receptor to initiate B cell response

15 Slide 43 ANTIBODY ACTIVITIES Neutralization Binds to toxins released by bacteria Hinders the ability of viruses to bind to cells Block antigen attachment (ie. taking parking space) Agglutination Bind to and clump bacteria Precipitation Render antigens insoluble Opsonization Coat antigen to promote ingestion and killing by phagocytic cells Complement Activation Combine with Ag on microbial surface to activate the complement cascade Slide 44 OTHER ANTIBODY ACTIVITIES Mucosal Protection IgA (sometimes IgG) inhibits pathogens from gaining attachment to mucosal surfaces Antibody dependent Cell Mediated Toxicity Bind to organisms via Fab region Allows attachment of NK to the Fc region Confer Immunity to a Fetus IgG passed across placenta Slide 45 IMMUNOGLOBULIN STRUCTURE 4 polypeptide chains 2 light chains 2 heavy chains Each chain has Variable region Specific for antigens Recognize and bind antigen Constant region Consistent among isotype

16 Slide 46 BASICS Why? How? Elements of the Immune System and Their Roles in Defense Origin of immune system components Antibody Structure Organs of the immune system Slide 47 ORGANS OF THE IMMUNE SYSTEM - REFERRED TO AS LYMPHOID ORGANS Slide 48 PRIMARY LYMPHOID ORGANS Sites of development and maturation Bone Marrow Contained in the hollow center of bones The source of all blood cells including T and B cells Site of B cell maturation Thymus Glandular structure located behind the breastbone above the heart Site of T cell maturation

17 Slide 49 SECONDARY LYMPHOID ORGANS Site of lymphocyte storage and Ag contact and response Located near potential portals of entry of pathogens Adenoids Lymph nodes Spleen Mucosa associated lymphoid tissue (MALT) Slide 50 BODY FLUID SYSTEMS Transport agents of the immune system Blood Lymph Flows through the lymphatic system Contains white blood cells, but very few or no red blood cells Slide 51 Kidney shaped organs located along lymphatic vessels Serve as filters for lymphatic fluid and storage sites for immune cells Contains specialized compartments to maximize interaction between immune cells and antigens LYMPH NODES

18 Slide 52 Acts like a large lymph node, but filters blood Collects bloodborne Ag Removes aged blood cells Serves to maximize interaction between immune cells/ components and antigens SPLEEN Slide 53 Slide 54 IMMUNITY

19 Slide 55 TYPES OF IMMUNITY Innate Immunity Genetically based and passed to offspring 1. Natural Surface barriers 2. Antigen, non-specific immune responses Acquired Immunity 3. Adaptive and specific immune responses Slide 56 BARRIERS Slide 57 SURFACE BARRIERS

20 Slide 58 NORMAL FLORA Microorganisms that live in and on the body to provide defense Skin Nasopharnyx Stomach & small intestine Large intestine The organisms of the normal flora work to destroy other organisms which attempt to invade their territory Slide 59 INNATE VS ADAPTIVE RESPONSE Slide 60 INNATE RESPONSE

21 Slide 61 INFLAMMATION Slide 62 MIGRATION Slide 63 EARLY INNATE RESPONSE Neutrophils and macrophages are key cells involved in the early response These cells migrate to the site and phagocytose the antigen

22 Slide 64 EARLY INNATE RESPONSE A small amt of antigen may be effectively eradicated A larger amt of antigen may require assistance Dendritic cells or macrophages present Ag to T-cells initiating an adaptive cellular response Slide 65 PASSIVE AND ACTIVE RESPONSE TO INVASION Non-Specific Defenses Physiological and Chemical Barriers Fever, cytokine release, complement cascade, macrophages, neutrophils, natural killer cells Specific Defenses Specific antibody and T cell response Passive Immunity Active Immunity Active Immunity Slide 66 ADAPTIVE RESPONSE

23 Slide 67 INITIATION OF CELL-MEDIATED IMMUNITY Macrophage or dendritic cell acts as an APC displaying a small amt of antigen on its surface via a MHC II protein Slide 68 BASIC CD4 + RESPONSE CD4 T cells which contains receptors specific for the antigen binds to the APC CD4 T Cells secretes cytokines Enhances macrophage phagocytosis Stimulate B cells to change to plasma cells Call in CD8 T cells and NK cells Slide 69 BASIC CD8 + RESPONSE CD8 cells which contains receptors specific for the antigen binds to the APC CD8 Cells Kill cells expressing appropriate Ag Release lymphotoxins which cause cell lysis Inhibit the production of cytotoxic T cells once they are no longer needed Down regulate the activity of other cells

24 Slide 70 BASIC CD8 + RESPONSE Slide 71 ACQUIRED, SPECIFIC IMMUNE RESPONSES Consists of antibodies and T-cells which contain receptors specific to invading antigens Upon first exposure to an antigen, a lag may exist in a specific immune response Upon re-exposure to an antigen, immune response is more rapid based on memory B and T cells Slide 72 IMMUNE RESPONSE Humoral Immunity Cell-mediated Immunity Antibody Cytotoxicity

25 Slide 73 B CELL ANTIBODY PRODUCTION B cell recognizes antigen which binds to immunoglobulin (Ig) receptors B cells internalize antigen and then act as APC to T- helper cells The T- helper cells secrete IL-4, IL-6 which activates the B cell to mature into a plasma cell Plasma cells produce antibody specific for that antigen Slide 74 HUMORAL IMMUNITY Primary component is antibody B cells evolve into plasma cells when signaled by T helper cells Antibody is produced from plasma cells Slide 75 PRIMARY & SECONDARY RESPONSE Primary Response Antibody production upon first exposure to a particular antigen takes 4 to 5 days A relatively small amount of IgM is produced that is short lived This is followed by IgG and IgA

26 Slide 76 SECONDARY RESPONSE B memory cells Upon re-exposure to an antigen, a larger amount of Ab is produced more rapidly which lasts for months or years The main type of Ab produced is IgG IgA and IgE may also be produced This is the basis for serial vaccination Slide 77 IS THERE SOMETHING WRONG WITH IGM? Slide 78 ANTIBODY FUNCTION Properties and IgM IgD IgG IgE IgA Functions Molecular weight (kda) % Serum 5-10 < < immunoglobulin Half-life (days) T-cell Independent Independent Dependent Dependent Dependent requirement Time/memory Early, primary Early, primary Later, memory Later, memory Later, memory Complement Opsonization Crosses placenta ++ Protects mucosa Activates mast +++ cell

27 Slide 79 HOW DOES THE IMMUNE SYSTEM BECOME MORE EFFICIENT? Slide 80 COMPLEMENT A group of 26 proteins produced by the liver and cells such as monocytes and macrophages Complement Cascade Ultimately leads to the formation of the membrane attack complex (MAC) punches holes in the cell wall of infected cells leading to death Slide 81 GENERAL COMPLEMENT ACTIVITIES Render bacteria more susceptible to phagocytosis Directly lyse some bacteria and foreign cells Increase vascular permeability Cause smooth muscle contraction promoting mast cell degranulation

28 Slide 82 ACTIVATION OF THE COMPLEMENT CASCADE Classical Pathway Initiated by immunologic means Alternate Pathway Lectin Pathway May be initiated without antibody present Slide 83 REACTION TO INFECTION Slide 84 Importance of Antimicrobial Defenses for Infectious Agents Host Defense Bacteria Intracellular Bacteria Viruses Fungi Parasites Complement Interferon-α/β ++++ Neutrophils Macrophages Natural killer cells +++ CD4 TH TH CD8 cytotoxic T lymphocytes Antibody (IgE)

29 Slide 85 BACTERIAL RESPONSE Slide 86 VIRAL RESPONSE Slide 87 ACUTE REACTION TO INFECTION A lag in antibody response may occur to a new type of antigen Cytokines are released early in the infection to initiate fever & the inflammatory response Some of these responses may be detrimental to the host ie. septic shock

30 Slide 88 PROPERTIES OF BACTERIA USED TO EVADE THE IMMUNE RESPONSE Structural features Adhesion features Secretion of Proteins and Toxins Slide 89 STRUCTURAL FEATURES Cell Wall Proteins Extracellular enzymes & toxins Slide 90 EXTRACELLULAR CAPSULES Protect the bacteria from phagocytosis

31 Slide 91 FORMATION OF BIOFILMS Slide 92 BINDING OF ANTIBODIES ie. Protein A produced by Staphylococcus aureus binds to the Fc portion of an antibody Slide 93 ADHESION Ability to adhere to the surface of human cells ie. Escherichia coli in the urinary tract

32 Slide 94 SECRETED PROTEINS Certain bacteria secrete cytolysins which lyse host cell membranes Slide 95 SECRETED PROTEINS ie. Hyaluronidase secreted by S. aureus Breaks down the hyaluronic acid matrix between cells Allows bacteria to penetrate skin more easily Slide 96 SECRETED PROTEINS Exotoxins May be produced by certain species of Gram positive and Gram negative bacteria Consist of A and B components May have various detrimental effects ie. Clostridium botulinum botulism caused by the toxin

33 Slide 97 SECRETED PROTEINS Endotoxins Produced by certain species of Gram negative bacteria Contained within the outer bacterial membrane May initiate the alternative complement pathway Results in uncontrolled release of cytokines and septic shock Slide 98 VIRAL INFECTION Immune system must destroy the host cell in order to destroy the viral cell Viruses utilized the immune system as a vehicle for infection ie. Human immunodeficiency virus Interferon is a key cytokine targeted against viruses Slide 99 USING THE IMMUNE SYSTEM COMPONENTS FOR TESTING Serologic Diagnosis

34 Slide 100 SEROLOGIC ASSAYS Complement fixation Hemagglutination inhibition Neutralization Immunofluorescence (direct and indirect) Latex agglutination In situ enzyme immunoassay (EIA) Enzyme-linked immunosorbent assay (ELISA) Radioimmunoassay (RIA) Slide 101 Specific Monoclonal Polyclonal Sensitive Reproducible results ANTIBODIES Slide 102 Direct Precipitation Diffusion Labeling Radioactive Fluoresence Enzymes Indirect Directed reactions ANTIBODY DETECTION

35 Slide 103 IMMUNOASSAYS Definition- An immunoassay is a biochemical test used to identify and quantify a specific antigen or antibody in a sample or solution through the use of an immunoglobulin. Immunohistology- Cell associated antigen Immunofluorescence and Enzyme immunoassay- EIA Direct Indirect Antibody and soluble antigen Enzyme Linked immunosorbent assay (ELISA) Western blot Complement Agglutination Slide 104 USES Phenotyping Diagnosis of infection Serotyping Transplantation

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