Cytotoxic NKG2C + CD4 T cells target oligodendrocytes in Multiple Sclerosis Fatma Zaguia Microbiology and Immunology McGill University, Montreal December 2012 A thesis submitted to McGill University in partial fulfillment of the requirements of the degree of Master’s in Science © Fatma Zaguia, 2012 Table of Contents ACKNOWLEDGEMENTS .................................................................................... 4 CONTRIBUTIONS OF AUTHORS ........................................................................ 8 ABSTRACT ....................................................................................................... 9 RÉSUMÉ ........................................................................................................ 10 INTRODUCTION AND LITTERATURE REVIEW ................................................... 12 Multiple Sclerosis .......................................................................................... 12 Disease Overview ................................................................................................. 12 Clinical Features ................................................................................................... 13 Pathological Features ........................................................................................... 13 Diagnosis and disease modulating therapies ......................................................... 14 Immune-mediated damage to the CNS .......................................................... 15 Evidence of autoimmunity in the pathogenesis of MS ........................................... 15 Immune mediators involved in disease pathogenesis ............................................ 16 CD8 T cells ................................................................................................................ 17 Natural Killer cells ..................................................................................................... 17 B cells ........................................................................................................................ 18 Gamma delta T cells ................................................................................................. 19 Macrophages ............................................................................................................ 20 Cytokines .................................................................................................................. 20 Role of CD4 T cells in MS pathogenesis ................................................................. 23 Implication in MS and EAE model ......................................................................... 23 Autoreactive CD4 T cells ........................................................................................ 25 Direct targeting of oligodendrocytes .................................................................... 26 Signaling through NKG2 receptors and the HLA-E ligand................................. 28 HLA-E ................................................................................................................... 28 NKG2A and NKG2C ............................................................................................... 30 Signaling through NKG2A ......................................................................................... 32 Signaling through NKG2C ......................................................................................... 33 FINDINGS ...................................................................................................... 36 Cytotoxic NKG2C + CD4 T cells target oligodendrocytes in Multiple Sclerosis.... 36 Abstract ............................................................................................................... 37 Introduction......................................................................................................... 38 Results and Discussion ......................................................................................... 40 Materials and Methods ........................................................................................ 49 Bibliography ........................................................................................................ 56 Figure legends ...................................................................................................... 60 2 DISCUSSION .................................................................................................. 70 CONCLUSION AND FUTUR DIRECTIONS .......................................................... 75 REFERENCES .................................................................................................. 77 3 ACKNOWLEDGEMENTS First and foremost I would like to thank my supervisors, Dr. Jack Antel and Dr. Nathalie Arbour. Dr. Antel, it has been an immense pleasure to have the opportunity to work in your laboratory. You have been an amazing mentor, teacher and a constant source of encouragement. No matter how busy your day was, you never hesitated to lean back in your chair, put your feet up on the table and tell me “OK…. Let’s take the time to discuss what’s next”. When times were tough, you always knew how to tell me to take a step back, and to remember the big picture and the story we want to tell. Science aside, you are one of the kindest people I have had the chance of meeting; thank you for always being so attentive and enthusiastic about whichever direction the project took. I can only start to show my gratitude by helping you from time to time navigate your Mac laptop. You have most definitely set an example as to the kind of clinician and researcher I aspire to become one day. My experience as a Master’s student would not h ave been complete without my co-supervisor, Dr. Nathalie Arbour. Nathalie, from the first day you sat me down to explain what a CD8 T cell was, I knew there was an encyclopedia of knowledge that I could gain from you. You really trained me from scratch, helped me overcome my fear of the FACS machine, and taught me the rigor and discipline required for “good science”. You have been incredibly generous with your time and attention throughout the years; hours were spent helping me with writing, presentations, understanding concepts – all this with incredible kindness and laughter. Any organizational skills that I possess today, I mostly have you to thank for (although my lab desk will probably always be just a little too messy for your liking…). Thank you both for an incredible MSc. experience! Next, I would like to thank members from the MNI laboratory. First, a big thank you to Manon Blain and Ellie McCrea, our two technicians and the “real 4 bosses” of the lab. You both make the environment and life in the lab so enjoyable and pleasant. Ellie, thank you for the hours spent helping me generate the T cell lines. We definitely had our shares of laughs and hair- tearing moments and no one activates those T cells better than you! Manon, “la maman du labo”, we would al l be lost without you. From day one I knew that when times were tough, there would always be someone to turn to for help or just a pep talk. A big thank you to Philippe Saikali, who started this project and who took time in the beginning of my MSc. to explain everything and anything; you were always an email away when I had questions and I am so grateful for that! Thank you Veronica Miron-Lafrance, Caroline Lambert and Qiao-Ling Cui for teaching me how to do the adults preps. I wish our paths had crossed more often; I know there is much to learn from you. Thank you Trina Johnson for your presence in lab meetings. You always say it as it is and some of the best constructive criticism has come from you. Big thanks also for hosting the greatest parties! And last but not least, I am eternally indebted towards Bryce Durafourt. Who knew that walking into the basement of the MNI on that first day I would be meeting one of my best friends and future classmates. Thank you for being an amazing friend throughout the years; you constantly pushed me, didn’t let me give up (and god knows there were times where that’s all I wanted to do!), took care of my traveling documents, put up with my hysterical moments (missing prereqs?!), corrected my statements (apparently I cannot use “whilst”), shared great Top40 music with me, and the list goes on….I can only hope that I’ve been half as valuable as a friend as you’ve been to me. Can’t wait until the days we “consult and discuss patient files while using our pagers”. 5 Now on to my lab at the CHUM. Thank you Diane Beauseigle for your kindness and help throughout the years. A big thank you to Alma Mohebiany. You were my first friend in the lab and here in Montreal; I am so happy that our friendship hasn’t wavered despite that you are now across the Atlantic Ocean. Thank you for sharing my love of music, baked sweets and large coffee mugs! One day we will find that perfect cup…Late days in the lab wouldn’t have been the same without you. Camille Pittet, thank you for being a great lab and life friend. Your organized, and well-written lab books made my life so easy on more than one occasion. You always had time for all my questions especially the eternal “what concentration did you use?”. And then we would proceed to go have lunch and talk about shoes; what more could I ask for? Thank you Raphael Schneider, for your help throughout my time in the lab. You always had a kind comment to say or a funny joke to crack; working together on the Journal Club paper was a pleasure. I would like to also