Washington University in St. Louis Washington University Open Scholarship Arts & Sciences Electronic Theses and Dissertations Arts & Sciences Spring 5-15-2020 Metabolic Requirements of NK Cell Responses to Viral Infection Annelise Yoo Mah-Som 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 Mah-Som, Annelise Yoo, "Metabolic Requirements of NK Cell Responses to Viral Infection" (2020). Arts & Sciences Electronic Theses and Dissertations. 2216. https://openscholarship.wustl.edu/art_sci_etds/2216 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 and Biomedical Sciences Immunology Dissertation Examination Committee: Megan Cooper, Chair Takeshi Egawa Todd Fehniger Anthony French Deborah Lenschow Wayne Yokoyama Metabolic Requirements of NK Cell Responses to Viral Infection by Annelise Yoo Mah-Som A dissertation presented to The Graduate School of Washington University in partial fulfillment of the requirements for the degree of Doctor of Philosophy May 2020 St. Louis, Missouri © 2020, Annelise Yoo Mah-Som Table of Contents List of Figures ................................................................................................................................ iv List of Abbreviations ....................................................................................................................... v Acknowledgments .......................................................................................................................... vi Abstract ........................................................................................................................................ viii Chapter 1: Introduction ................................................................................................................... 1 1.1 Natural Killer Cells and Their Functions ......................................................................... 3 1.2 Metabolic Regulation of Immune Cells ............................................................................ 7 1.2.1 Major Metabolic Pathways .................................................................................................... 7 1.2.2 Metabolic Regulation in T cells ........................................................................................... 11 1.2.3 Metabolic Regulation in Macrophages ................................................................................ 13 1.2.4 Metabolic Regulation in Dendritic Cells ............................................................................. 16 1.2.5 Metabolic Regulation in B cells .......................................................................................... 17 1.2.6 Metabolic Regulation in Neutrophils .................................................................................. 19 1.3 Metabolic Regulation in NK cells .................................................................................. 20 1.4 Conclusions and Aims .................................................................................................... 28 1.5 References ...................................................................................................................... 31 Chapter 2: Glycolytic Requirement for NK Cell Cytotoxicity and Cytomegalovirus Control ..... 38 2.1 Abstract ........................................................................................................................... 39 2.2 Introduction .................................................................................................................... 39 2.3 Results ............................................................................................................................ 42 2.4 Discussion ....................................................................................................................... 58 2.5 Materials and Methods ................................................................................................... 63 2.6 Supplementary Figures ................................................................................................... 69 2.7 References ...................................................................................................................... 73 Chapter 3: Cox10 is Required for Expansion of Ly49H+ NK cells During Murine Cytomegalovirus Infection ............................................................................................................ 78 3.1 Abstract ........................................................................................................................... 79 3.2 Introduction .................................................................................................................... 80 3.3 Results ............................................................................................................................ 82 3.4 Discussion ....................................................................................................................... 92 ii 3.5 Materials and Methods ................................................................................................... 96 3.6 Supplementary Figures ................................................................................................. 101 3.7 References .................................................................................................................... 102 Chapter 4: Conclusions and Future Directions ........................................................................... 105 4.1 Glycolytic Metabolism in NK Cell Antiviral Function ................................................ 106 4.2 Oxidative Metabolism in NK Cell Antiviral Function ................................................. 108 4.3 NK Cells in the Immunometabolism Paradigm ............................................................ 110 4.4 Future Directions .......................................................................................................... 112 4.5 References .................................................................................................................... 116 iii List of Figures Figure 1.1: NK cell functions during MCMV infection.................................................................6 Figure 1.2: Major metabolic pathways and programs in immune cells..........................................8 Figure 1.3: Metabolic regulators.....................................................................................................9 Figure 1.4: Mitochondrial electron transport chain.......................................................................10 Figure 1.5: Increased rates of metabolism with prolonged IL-15 exposure but not short-term activation...................................................................................................................22 Figure. 1.6: Naive NK cells require OXPHOS for receptor-stimulated IFN-g production, but activation with IL-15 eliminates this requirement................................................................23 Figure 2.1: Glucose metabolism blockade during activation decreases NK cell proliferation and cytotoxicity...........................................................................................................................43 Figure 2.2: In vitro treatment with 2DG decreases NK cell production of granzyme B and alters actin accumulation at the immunological synapse............................................................45 Figure 2.3: 2DG treatment in vivo causes decreased target clearance during MCMV infection..47 Figure 2.4: 2DG treatment in vivo confers MCMV susceptibility................................................50 Figure 2.5: NK activation with ALT-803 rescues MCMV susceptibility caused by 2DG treatment.....................................................................................................................................52 Figure 2.6: ALT-803 rescue of 2DG-treated mice requires NK cells, and ALT-803 priming can eliminate susceptibility to MCMV caused by mTOR inhibition.........................................54 Figure 2.7: 2DG decreases human NK cell cytotoxicity...............................................................56 Figure 2.8: ALT-803 treatment of CMV reactivation in a post-hematopoietic cell transplant patient..........................................................................................................................................58 S. Figure 2.1: Effects of 1mM 2DG culture on NK cell metabolism and killing..........................69 S. Figure 2.2: T cells upregulate Gzmb and proliferate, but are not affected by 2DG treatment..70 S. Figure 2.3: IFN-γ production by NK cells during MCMV infection is not inhibited by 2DG..71 S. Figure 2.4: MCMV susceptibility is dependent on the dose of 2DG and the inoculum of MCMV........................................................................................................................................71 S. Figure 2.5: The effects of ALT-803 are not mediated