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Evaluation of IL2 and HLA on the Homeostasis and Function of Human CD4 and CD8 T Cells

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Evaluation of IL2 and HLA on the Homeostasis and Function of Human CD4 and CD8 T Cells University of Massachusetts Medical School eScholarship@UMMS GSBS Dissertations and Theses Graduate School of Biomedical Sciences 2017-09-15 Evaluation of IL2 and HLA on the Homeostasis and Function of Human CD4 and CD8 T Cells Philip A. Durost University of Massachusetts Medical School Let us know how access to this document benefits ou.y Follow this and additional works at: https://escholarship.umassmed.edu/gsbs_diss Part of the Immunity Commons Repository Citation Durost PA. (2017). Evaluation of IL2 and HLA on the Homeostasis and Function of Human CD4 and CD8 T Cells. GSBS Dissertations and Theses. https://doi.org/10.13028/M2C979. Retrieved from https://escholarship.umassmed.edu/gsbs_diss/936 This material is brought to you by eScholarship@UMMS. It has been accepted for inclusion in GSBS Dissertations and Theses by an authorized administrator of eScholarship@UMMS. For more information, please contact [email protected]. EVALUATION OF IL2 AND HLA ON THE HOMEOSTASIS AND FUNCTION OF HUMAN CD4 AND CD8 T CELLS A Dissertation Presented By PHILIP A DUROST Submitted to the Faculty of the University of Massachusetts Graduate School of Biomedical Sciences, Worcester in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY SEPTEMBER 15, 2017 IMMUNOLOGY AND MICROBIOLOGY PROGRAM EVALUATION OF IL2 AND HLA ON THE HOMEOSTASIS AND FUNCTION OF HUMAN CD4 AND CD8 T CELLS A Dissertation Presented By PHILIP A DUROST This work was undertaken in the Graduate School of Biomedical Sciences Immunology and Microbiology Program Under the mentorship of Michael Brehm, Thesis Advisor The signatures of the Dissertation Defense Committee signify completion and approval as to style and content of the Dissertation Dale Greiner, Member of Committee Anne Rittenhouse, Member of Committee John Harris, Member of Committee Evan Jellison, External Member of Committee The signature of the Chair of the Committee signifies that the written dissertation meets the requirements of the Dissertation Committee Ann Rothstein, Chair of Committee The signature of the Dean of the Graduate School of Biomedical Sciences signifies that the student has met all graduation requirements of the School. Anthony Carruthers, Ph.D., Dean of the Graduate School of Biomedical Sciences Abstract Homeostasis of human T cells is regulated by many factors that control proliferation, differentiation of effector cells and generation of memory. Our current knowledge of the mechanisms controlling human T cell homeostasis in vivo is based on experiments in small animal models. However many differences exist between immune systems of mice and humans, including cell composition, function, and gene expression. Humanized mouse models have shown great value in the study of human immunobiology. I have used novel humanized mouse models to examine the role of human MHC (HLA) and human IL2 in CD8 T cell and CD4 regulatory T cell (Treg) homeostasis. To study human CD8 T cells I engrafted CD8 T cells from healthy donor PBMC into NOD-scid IL2rgnull (NSG) mice that lacked expression of murine MHC and that expressed HLA-A2. My data demonstrate that CD8 T cell survival and effector function required the presence of HLA- A2, helper function from human CD4 T cells and exogenous human IL2. To study human Treg homeostasis I used NSG mice engrafted with human fetal thymus and hematopoietic stem cells (BLT model). NSG-BLT mice support the growth of human thymic tissue and enable the efficient development of HLA-restricted Treg and conventional T cells. Using an AAV vector to express human IL2, I demonstrated that functional human Treg but not conventional T cells increased in number in NSG-BLT mice and that this coincided with increases in activated human NK cells. Overall my research has revealed that HLA and human IL2 have an essential role in human T cell survival and function in vivo. ( Table of contents #$"$************************************************************************************************************************************* #$ #****************************************************************************************************************************& #$ %"#**************************************************************************************************************************& #$ !("$$"!" %($%$ "*********************************************& "&$ #**************************************************************************************************************************' !$" - $" %$ ********************************************************************************************************/ Part 1: Innate immunity, T cell types and their respective roles role in the immune response*************************************************************************************************************************************************1 Innate immunity222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222: Adaptive immunity222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222287 Part 2: The development, homeostatic proliferation and the function of T cells****************/7 Development of T cells in the Thymus and Central Tolerance2222222222222222222222222222222222222222222222222222228@ Treg Development in the Thymus and the Periphery2222222222222222222222222222222222222222222222222222222222222222222222229= Peripheral Homeostasis and Survival of Tconv and Treg22222222222222222222222222222222222222222222222222222222222222229> The migratory path of naïve T cells and the initiation of the T cell response to foreign antigen.222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222229? Memory T cell development22222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222229? Part 3: Graft versus host disease***************************************************************************************************07 GVHD is a Complication of Tissue Transplantation2222222222222222222222222222222222222222222222222222222222222222222222222:7 The Phases in the Development of acute GVHD22222222222222222222222222222222222222222222222222222222222222222222222222222222:8 Chronic and mixed forms of GVHD2222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222:9 Part 4: Gene therapy************************************************************************************************************************11 A Complicated History With Mixed Results2222222222222222222222222222222222222222222222222222222222222222222222222222222222222222:: Viral Vectors as Adaptive Gene-Delivery Tools222222222222222222222222222222222222222222222222222222222222222222222222222222222:= Part 5: The development and application of the humanized mouse model*************************15 Mutations and Differences in the Lineages of Mice Used in Humanized Mouse Models2222:> Models of the Human Immune system222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222;7 !$" - $"$$%$ %6# %+ + ***************************************************************************************************23 Introduction***************************************************************************************************************************************23 Materials and methods********************************************************************************************************************25 Results*************************************************************************************************************************************************3. CD8 T cells engraft in NSG mice2222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222222<7 Engrafted CD8 T cells mediate a GVHD-like response and migrate into the host tissues222<> CD8 T cell engraftment and function depends upon the expression of MHC-I22222222222222222222222=< 3 !& *#$%%! 39("#!"! 22222222222222222222>: #%## *****************************************************************************************************************************************57 !$" - %($ # $"$$)#%"&&)%$ %6#***********************************************************************************************************6/ Introduction: Cellular and cytokine factors that regulate T cell survival and function***6/ Materials and methods********************************************************************************************************************60 ( Results*************************************************************************************************************************************************63 Purified human CD8 T cells are dependent upon other cell types to successfully engraft222?< Titration of CD8p cells into NSG mice reveals that there is a threshold for engraftment22222@8 Mice expressing transgenic human IL-7 fail to engraft with purified CD8 T cells22222222222222222@< Exogenous human IL-2 can support the engraftment of purified CD8 T cells2222222222222222222222222@? #%## **************************************************************************************************************************************/./ !$" -"%$ "($ % +0 " #$##%$ ******************************************************************************************************************/.1 Background
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