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  • Mechanisms of early immune activation in response to

    hepatitis B virus

    Sílvia Margarida Teixeira Vilarinho

    Dissertação de doutoramento em Ciências Biomédicas

    2008

  • Sílvia Margarida Teixeira Vilarinho

    Mechanisms of early immune activation in response to

    hepatitis B virus

    Dissertação de Candidatura ao grau de Doutor em Ciências Biomédicas submetida ao Instituto de Ciências Biomédicas de Abel Salazar da Universidade do Porto Orientador – Dr. Jody L. Baron, Assistant Professor of Medicine, University of California San Francisco Co-orientador – Professor Doutor Rui Appelberg, Professor Catedrático, Instituto de Ciências Biomédicas de Abel Salazar da Universidade do Porto

  • This dissertation is dedicated to the memory of my grandmother Emília Brandão,

    who passed away victim of a liver-related disease

    while I was performing these studies

  • i

    Table of Contents

    ABBREVIATIONS .......................................................................................................................................V

    ACKNOWLEDGMENTS............................................................................................................................VII

    ABSTRACTS .............................................................................................................................................. IX

    ABSTRACT ..............................................................................................................................................XI RESUMO................................................................................................................................................ XIII RÉSUMÉ .................................................................................................................................................XV

    CHAPTER I - INTRODUCTION .................................................................................................................. 1

    HEPATITIS B VIRUS .................................................................................................................................... 3 THE LIVER AS AN IMMUNOLOGICAL ORGAN.................................................................................................. 6 HEPATITIS B AS AN IMMUNOLOGICAL DISEASE .......................................................................................... 11 OVERVIEW OF THE MODELS USED TO STUDY HBV IMMUNOPATHOGENESIS .............................................. 14 DISEASE MODEL: TRANSGENIC MOUSE MODEL OF PRIMARY HEPATITIS B VIRUS INFECTION ...................... 18 NATURAL KILLER T CELLS AND THEIR FUNCTION ...................................................................................... 21 NATURAL KILLER CELLS AND THEIR FUNCTION.......................................................................................... 24 NKG2D RECEPTOR ................................................................................................................................. 26 NKG2D LIGANDS..................................................................................................................................... 29 NKG2D RELATED IMMUNE DISEASES ....................................................................................................... 31 NATURAL KILLER T CELLS AND HEPATITIS ................................................................................................ 33 OBJECTIVES............................................................................................................................................. 35

    CHAPTER II - BLOCKADE OF NKG2D ON NKT CELLS PREVENTS HEPATITIS AND THE ACUTE IMMUNE RESPONSE TO HEPATITIS B VIRUS.................................................................................... 37

    INTRODUCTION ................................................................................................................................... 41 RESULTS............................................................................................................................................... 42 DISCUSSION......................................................................................................................................... 54

    CHAPTER III - THE ROLE OF DAP10 AND DAP12 IN NKG2D-MEDIATED NKT CELL ACTIVATION.............................................................................................................................................. 57

    INTRODUCTION ................................................................................................................................... 59 RESULTS............................................................................................................................................... 60 DISCUSSION......................................................................................................................................... 63

    CHAPTER IV - MILD HEPATIC NECROSIS DETECTED IN HBV-TRANSGENIC RAG-1-/- MICE IS IFNG AND NK CELL-INDEPENDENT..................................................................................................... 67

    INTRODUCTION ................................................................................................................................... 69 RESULTS............................................................................................................................................... 70 DISCUSSION......................................................................................................................................... 78

    CHAPTER V - DISCUSSION AND CONCLUSIONS.............................................................................. 81

    INNATE AND “INNATE-LIKE” IMMUNE RESPONSES TO HBV-EXPRESSING CELLS ......................................... 83 HBV IMMUNOPATHOGENESIS: LESSONS IN NKT CELL BIOLOGY ............................................................... 86 TRANSGENIC MOUSE MODEL OF PRIMARY HUMAN HBV INFECTION........................................................... 91 LIMITATIONS OF OUR TRANSGENIC MOUSE MODEL OF PRIMARY HUMAN HBV INFECTION .......................... 93 HBV TREATMENT AND THERAPIES: NEW INSIGHT ..................................................................................... 94 NKG2D AND OTHER IMMUNE-MEDIATED LIVER DISEASES......................................................................... 95 FINAL CONSIDERATIONS ........................................................................................................................... 95

    CHAPTER VI - FUTURE PERSPECTIVES ............................................................................................. 97

    RAE-1 REGULATION WITHIN THE LIVER .................................................................................................... 99 DAP10 AND DAP12 IN NKT CELL ACTIVATION...................................................................................... 100

  • ii

    HUMAN STUDIES..................................................................................................................................... 100

    CHAPTER VII - MATERIALS AND METHODS .................................................................................... 101

    APPENDIX 1 ............................................................................................................................................ 109

    EPILOGUE ............................................................................................................................................... 113

    REFERENCES......................................................................................................................................... 115

  • iii

    List of Figures Figure 1.1……………………………………………………………………………………......6

    Figure 1.2……………………………………………………………………………………......8

    Figure 1.3.………………………………………………………………………………………19

    Figure 1.4.………………………………………………………………………………………28

    Figure 1.5……………………………………………………………………………………….30

    Figure 2.1……………………………………………………………………………………….43

    Figure 2.2……………………………………………………………………………………….45

    Figure 2.3……………………………………………………………………………………….47

    Figure 2.4……………………………………………………………………………………….50

    Figure 2.5……………………………………………………………………………………….51

    Figure 2.6……………………………………………………………………………………….52

    Figure 2.7……………………………………………………………………………………….53

    Figure 3.1……………………………………………………………………………………….60

    Figure 3.2……………………………………………………………………………………….61

    Figure 3.3…………………………