1. MICR 304 Immunology & Serology Lecture 6 NK Cells, Lymphocytes Chapter 1.4 –1.17; 2.30 – 2.33 Lecture 6 NK Cells, Lymphocytes Chapter 1.4 –1.17; 2.30 – 2.33

2. Overview of Today’s Lecture Lymphoid cell lineage NK cells and their receptors Lymphocytes Principles of cell signaling

3. Key Players in Immunology InnateAdaptive Cells Phagocytes Epithelial Cells NK Cells Lymphocytes (B-Ly, T-Ly) Defense Effector Molecules Complement Antimicrobial (Poly)Peptides Antimicrobial lipids? Antibodies

4. Lymphoid Cell Lineage Naïve Lymphocytes Effector Lymphocytes IL15IL2, IL7 NK cells

5. NK-Cells Form Part of the Innate Immune System Large granular lymphoid cells Develop in bone marrow (and maybe in the thymus?) –IL-15 dependent Circulate in peripheral blood Kill infected and tumor cells –Well documented role in defense against infections with viruses from Herpes family –Infections with Leishmania and Listeria –Tumor cells in vitro Invariant receptors

6. Infections with Documented NK Cell Mediated Defense Herpes simplex virus Cutaneous leishmaniosis (“Bhagdad Sore”) Cytomegalovirus (CMV)

7. Killing of Tumor Cells by NK Cells in vitro Tumor Cell NK-Cell t0 t60

8. NK Cells are an Early Component of Host Defense against Viral Infections Virus titer levels Virus titer decreases

9. Contributions of NK Cells to Defense against Virus Infections NK cells control the initial infection until adaptive immune system can eliminate the virus NK cells are activated by cytokines made by infected cells, macrophages and dendritic cells –INF , IFN , IL12, TNF , NK cells begin to kill infected cells In addition, NK cells secrete IFN  activating macrophages and dendrictic cells, and driving immune response to TH1 type response NK-DC cross talk

10. How do NK Cells Kill? Triggered by two mechanisms –Antibody dependent cytotoxicity –Recognition of altered surface molecules Mediated by: –Perforin Pore-forming toxin Permeabilizes target cell membrane –Granzymes Enzymes Induces apoptosis (cell suicide) –TNF  Apoptosis Packaged in Granules

11. Antibody Dependent Cellular Cytotoxicity (ADCC)  NK cells recognize antibodies that are bound to target cells through Fc  III receptor (CD16)  CD16 is an activating NK cell receptor

12. NK Cells Recognize Altered Cell Surface Depends on two types of receptors: –Activating (trigger cytotoxicity) Direct recognition of changes in cell surface glycoproteins –Inhibitory Normal MHC I Balance of activating and inhibitory receptor signaling regulates NK cell responses In normal cells, inhibitory receptors override activating signals

13. NK Cell Activation and Inhibition NK and normal cell: Inhibition > activation NK and abnormal cell: Activation uninhibited

14. Activating Receptors on NK Cells Direct recognition of changes in cell surface glycoprotein Short cytoplasmic tail Associate with signaling adaptor proteins that initiate activation cascade Example: NKG2D –Binds to proteins that are expressed in cellular stress

15. Inhibitory Receptors on NK Cells Specific for various MHC I molecules –If MHC I is altered or down regulated inhibitory receptors do not bind and NK cells become uninhibited Long cytoplasmic tail Associates with inhibitory phosphatases Example: KIR-2DL

16. Additonal Roles of NK Cell in our Immune System Recent evidence suggests that NK cells contribute to the pathogenesis of hypersensitivity and autoimmune diseases (Bubnoff et al., 2010) –Cytokines released by NK cells influence disease development.

17. Active Learning Both neutrophils and NK cells can recognize their target cells through antibodies. In what are their responses to the target cells different?

18. Adaptive Immunity

19. Lymphocytes Mediate adaptive immune responses + Specific antigen recognition Clonal proliferation m Memory m m m m m m

21. Lymphocyte Development Lymphoid Progenitor Helper Cytotoxic Regulatory NK-Cells Plasma cell B-Lymphoblast Thymocyte T-Lymphoblast Prolymphocyte B-Lymphocyte T lymphocyte IL2, IL7 IL15

22. Adaptive Immune Responses Antibody secretion (B-cells) –To block the invader –To enhance phagocyte and NK cell function Activation of effector cells via cytokines (TH cells) –to better fight infection Destruction of uncontrollable cells (CTL) –Infection by intracellular pathogens –Tumor cells Control of immune response (Regulatory T cells)

23. Lymphocytes Monitor the Body

24. Dendritic Cells Initiate Adaptive Immune Responses

25. Clonal Expansion of Activated Lymphocytes

26. From Antigen to Response Specific antigen recognition Common signal transduction nucleus Reaction

27. General Principle of Antigen Receptors variable constant Cytoplasmic tail Receptor Signal transducer Cytoplasmic tail Intracellular cascade of signal transduction

28. Each Lymphocyte Carries only One Type of Receptor “Billions of lymphocytes collectively carry millions of antigens”

29. Today’s Take Home Message The lymphoid cell lineage gives rise to lymphocyte (IL2 and IL7 dependent) and to NK cells (IL15 dependent) NK cells kill infected and tumor cells are the first producers of IFN  during an immune response. NK cells are regulated by a complex balance of activating and inhibitory receptors. Activating receptors like NKG2D recognize new surface molecules produced upon cellular stress. Inhibitory NK cell receptors like KIR-2DL bind to normal MHC I molecules. Lymphocytes express antigen specific receptors and unlike other immune cells they undergo clonal expansion and development into memory cells.

30. Additional Resources Zhou F.(2010) Expression of multiple granzymes by cytotoxic T lymphocyte implies that they activate diverse apoptotic pathways in target cells. Int Rev Immunol. 2010;29(1):38-55. von Bubnoff et al. (2010) Natural killer cells in atopic and autoimmune diseases of the skin. J Allergy Clin Immunol. Jan;125(1):60-8. http://education.vetmed.vt.edu/curriculum/vm8054/Labs/Lab6/I MAGES/MONOCYTE%20IN%20SMEAR.JPG