Washington University in St. Louis Washington University Open Scholarship Arts & Sciences Electronic Theses and Dissertations Arts & Sciences Spring 5-15-2016 The Resolution Phase of NK Cell Proliferation and IFN Production Following Viral Infection Are Highly Regulated Processes. Leslie Abigail Fogel Washington University in St. Louis Follow this and additional works at: https://openscholarship.wustl.edu/art_sci_etds Part of the Allergy and Immunology Commons, Immunology and Infectious Disease Commons, and the Medical Immunology Commons Recommended Citation Fogel, Leslie Abigail, "The Resolution Phase of NK Cell Proliferation and IFN Production Following Viral Infection Are Highly Regulated Processes." (2016). Arts & Sciences Electronic Theses and Dissertations. 760. https://openscholarship.wustl.edu/art_sci_etds/760 This Dissertation is brought to you for free and open access by the Arts & Sciences at Washington University Open Scholarship. 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LOUIS Division of Biology & Biomedical Sciences Immunology Dissertation Examination Committee: Anthony French, Chair Paul Allen Marco Colonna Todd Fehniger John Russell Wayne Yokoyama The Resolution Phase of NK Cell Proliferation and IFNγ Production Following Viral Infection Are Highly Regulated Processes. by Leslie Abigail Fogel A dissertation presented to the Graduate School of Arts & Sciences of Washington University in partial fulfillment of the requirements for the degree of Doctor of Philosophy May 2016 St. Louis, Missouri © 2016, Leslie Abigail Fogel Table of Contents List of Figures...............................................................................................................................iii List of Tables ................................................................................................................................ iv List of Abbreviations .................................................................................................................... v Acknowledgments ........................................................................................................................ ix Abstract......................................................................................................................................... xi Chapter 1: Introduction ............................................................................................................... 1 Figures....................................................................................................................................................... 9 References............................................................................................................................................... 11 Chapter 2: Markers of Nonselective and Specific NK Cell Activation.................................. 19 Abstract................................................................................................................................................... 19 Introduction............................................................................................................................................. 20 Materials and Methods............................................................................................................................ 23 Results..................................................................................................................................................... 26 Discussion............................................................................................................................................... 33 Figures..................................................................................................................................................... 37 References............................................................................................................................................... 50 Chapter 3: The resolution of NK cell proliferation depends on αβ and γδ T cells ............... 57 Abstract................................................................................................................................................... 57 Introduction............................................................................................................................................. 58 Materials and Methods............................................................................................................................ 59 Results..................................................................................................................................................... 66 Discussion............................................................................................................................................... 72 Figures..................................................................................................................................................... 76 Acknowledgments................................................................................................................................... 87 Supplemental Tables............................................................................................................................... 88 References............................................................................................................................................. 108 Chapter 4: Apoptosis mediates the resolution of NK cell production of IFNγ following infection...................................................................................................................................... 115 Abstract................................................................................................................................................. 115 Introduction........................................................................................................................................... 116 Materials and Methods.......................................................................................................................... 118 Results................................................................................................................................................... 121 Discussion............................................................................................................................................. 122 Figures................................................................................................................................................... 125 References............................................................................................................................................. 130 Chapter 5: Future Experiments .............................................................................................. 133 Curriculum vitae....................................................................................................................... 136 ii List of Figures Figure 1.1: The extrinsic and intrinsic pathways of apoptosis. 9 Figure 2.1: Sca-l is upregulated on NK cells 2 days p.i. MCMV to a greater extent than 37 other identified activation markers. Figure 2.2: Sca-1 and CD69 expression correlate with IFN-γ production. 39 Figure 2.3: Expression of KLRG1, CD27, and Sca-1 is differentially regulated on 40 Ly49H+ and Ly49H- NK cells during MCMV infection. Figure 2.4: Differential expression of KLRG1, CD27, and Sca-1 on Ly49H+ and Ly49H- 42 NK cells during MCMV infection depends on Ly49H recognition of m157 protein. Figure 2.5: Proliferation alone does not result in the differential expression of KLRG1, 44 CD27, or Sca-1. Supplemental Figure 2.1: Marker expression on and IFN-γ production by hepatic NK 45 cells following MCMV infection. Supplemental Figure 2.2: Viral load does not affect the differential expression of 47 KLRG1, CD27, and Sca-1 on Ly49H+ and Ly49H- NK cells during MCMV-Δm157 infection. Figure 3.1: There is prolonged proliferation of NK cells in RAG-/- mice. 76 Figure 3.2: Bim regulates contraction of NK cells but not the resolution of NK cell 78 proliferation. Figure 3.3: Both αβ and γδ T cells have a redundant ability to resolve NK cell 79 proliferation. Figure 3.4: NK/T cell co-culture confirms that activated T cells inhibit NK cell 81 proliferation. Figure 3.5: Adoptive transfer of T cells into TCRβ/δ-/- mice to determine the 82 requirements for the resolution of NK cell proliferation. Figure 3.6: Analysis of gene expression in naïve CD8+ T cells compared to CD8+ and γδ 84 T cells from mice 5 days p.i. with MCMV. Figure 3.7: Adoptive transfer of T cells into TCRβ/δ-/- mice to test the requirement of 86 genes identified in the microarray. Figure 4.1: Time course of NK cell responses to MCMV. 125 Figure 4.2: IFNγ production by NK cells is associated with apoptosis. 126 Figure 4.3: TRAIL-R, but not Fas, is required for the resolution of IFNγ production by 128 NK cells. iii List of Tables Table 2.1: Marker expression on NK cells from naïve mice or from mice 1.5 or 2 days 49 p.i. MCMV Supplemental Table 3.I: List of genes upregulated at least 3 fold in only MCMV- 88 specific CD8+ T cells and γδ T cells compared to naïve CD8+ T cells Supplemental Table 3.II: List of genes upregulated at least 3 fold in only MCMV- 92 specific