Scott Abrams, Ph.D. Professor of Oncology, x4375 Kuby Immunology SEVENTH EDITION

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1 Scott Abrams, Ph.D. Professor of Oncology, x4375 Kuby Immunology SEVENTH EDITION CHAPTER 13 Effector Responses: Cell- and Antibody-Mediated Immunity Copyright 2013 by W. H. Freeman and Company

2 Fundamental Distinctions Between Humoral and Cell- Mediated Immune Responses recognition of extracellular versus intracellular pathogens cell surface vs. MHC-peptide complexes CMI primarily effective against: virus-infected cells intracellular bacteria, fungi tumors graft rejection 2

3 The Central Players of the Cell-Mediated Immune Response adaptive immunity: specific (Ag-specific, MHCrestricted) CD8 + CTLs CD4 + T h 1 (mediate DTH and recruitment of additional effectors, mainly macrophages, to sites of inflammation) CD4 + T h 2 (promote antibody production) innate immunity: nonspecific lymphoid: NK cells, NKT cells non-lymphoid: macrophages, granulocytes (i.e., neutrophils, eosinophils) 3

4 Cell-Mediated Effector Responses 4

5 Generation of Effector CD8 + CTLs 5

6 Generation of Effector CD8 + CTL (cont d) 6

7 CTL-Tumor Cell Interaction 7

8 Steps of CTL-Mediated Target Cytolysis 8

9 Early Stages of the CTL-Target Interaction: TCR Activation Facilitates Conformational Changes in LFA-1 Which Increase Affinity for ICAM-1 9

10 Later Stages of the CTL-Target Interaction: Pore Formation in the Target Cell Membrane 10

11 Experimental Evidence for Perforin and Fas Ligand as the Major CTL Effector Mechanisms 11

12 Apoptotic Pathways Triggered by Perforin and FasL-Based Effector Mechanisms 12

13 Assays for Detection and Measurement of T Cell Responses 13

14 Proliferation as a Measurement of T Cell Response 14

15 Cytotoxicity as a Measurement of T Cell Response 15

16 Phenotypic Analysis of MHC-Tetramer -Reactive Cells as a Measurement of T Cell Response 16

17 Tracking Ag-Specific CTLs In Vivo Using MHC-Tetramers 17

18 Innate Immunity NK cells NKT cells phagocytes macrophages granulocytes 18

19 Natural Killer Cells activity due to large granular lymphocytes, which comprise 5 10% of PBMC important for host defense against certain intracellular pathogens, including bacteria, viruses and tumor cells also, important for immune regulation and bridging innate and adaptive immune responses via cytokine production, such as IFN-γ modulate macrophage effector functions influence T h 1 commitment involved in early phase of infections, which provide ample kinetics for CTL generation; NK cells are stimulated by; e.g., IFN-α, IL-2, IL-12 lytic mechanisms are similar to those used by CTL perforin pathway is constitutively active death receptor killing cytotoxic cytokines immune response is dictated by the balance of positive and negative signals delivered via activating and inhibitory receptors 19

20 Model of Cross-Talk Between NK (Innate) and Adaptive Immunity in Response to a Viral Challenge 20

21 NK Recognition Activating Receptors (AR) interact with stress-induced ligands induced on target cells as a result of DNA damage, infection, etc., leading to downregulation of conventional MHC molecules induction of stress-induced ligands in concert with the loss of MHC expression culminates in NK activation Inhibitory Receptors (IR) interact with MHC molecules signals generated via IR inhibit NK activation under normal conditions of no cell damage under conditions of cell damage if MHC expression is not profoundly compromised IR signals predominate over AR signals 21

22 Model for NK Recognition and Lysis of Target Cells: Opposing-Signals Model AR engage ligands on target cell surfaces activation can be circumvented by IR which detect MHC expression levels NK activity is suppressed if MHC levels are unaltered, thus sparing normal cells 22

23 Experimental Design to Test for NK Cell Memory 23

24 Evidence for NK Cell Memory Response 24

25 Summary: Comparison Between NK Cells and CTLs Similarities derived from bone marrow and common early progenitors express shared markers, such as CD2, IL-2Rβ share lytic pathways, including perforin and FasL evidence for immunologic memory Differences do not express Ag-specific TCR or CD3 are not MHC-restricted do not require antigenic priming develop extra-thymically: can be found in nude, SCID and RAG-deficient mice, all of which lack functional T cells 25

26 Summary: Comparison Between NKT Cells and CTLs Similarities derived from bone marrow and common early progenitors, including thymic development express conserved elements of the TCR-α/β chains, and some subsets may express CD4 thought to be involved in the control of certain bacterial and viral infections, as well as tumors, and does so via cytokines (e.g., TNF-α) or Fas/FasL interactions Differences are not conventionally MHC-restricted, but recognize glycolipids presented by a non-polymorphic CD1d molecule currently, no evidence for immunologic memory express markers more characteristic of NK cells than T cells 26

27 Summary: Innate and Adaptive Immune Mechanisms of Antigen Recognition 27

28 Summary: Cell-Mediated Effector Mechanisms 28

29 Antibody-Mediated Effector Functions, Including ADCC 29

30 Antibody-Mediated Effector Functions 30

31 Signal Transduction Pathways Triggered in Response to FcR Engagement 31

32 Antibody-Dependent Cellular Cytotoxicity - NK cells, monocytes/macrophages, neutrophils and eosinophils express Fc receptors reactive with the Fc portion of target cell surfacespecific IgG - multiple mechanisms of lysis, including degranulation, seceretion of cytotoxic enzymes/cytokines 32

33 33

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