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Interleukin-21 in Immunity and Autoimmunity

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Interleukin-21 in Immunity and Autoimmunity Interleukin-21 in immunity and autoimmunity Alexis Vogelzang A thesis submitted for the degree of Doctor of Philosophy in the Faculty of Medicine, University of New South Wales Mucosal Autoimmunity Unit, Garvan Institute of Medical Research Sydney, Australia Awarded September 2010 1 ORIGINALITY STATEMENT ‘I hereby declare that this submission is my own work and to the best of my knowledge it contains no materials previously published or written by another person, or substantial proportions of material which have been accepted for the award of any other degree or diploma at UNSW or any other educational institution, except where due acknowledgement is made in the thesis. Any contribution made to the research by others, with whom I have worked at UNSW or elsewhere, is explicitly acknowledged in the thesis. I also declare that the intellectual content of this thesis is the product of my own work, except to the extent that assistance from others in the project's design and conception or in style, presentation and linguistic expression is acknowledged.’ Signed …………………………………………….............. Alexis Vogelzang Date …………………………………………….............. 2 COPYRIGHT STATEMENT ‘I hereby grant the University of New South Wales or its agents the right to archive and to make available my thesis or dissertation in whole or part in the University libraries in all forms of media, now or here after known, subject to the provisions of the Copyright Act 1968. I retain all proprietary rights, such as patent rights. I also retain the right to use in future works (such as articles or books) all or part of this thesis or dissertation. I also authorise University Microfilms to use the 350 word abstract of my thesis in Dissertation Abstract International (this is applicable to doctoral theses only). I have either used no substantial portions of copyright material in my thesis or I have obtained permission to use copyright material; where permission has not been granted I have applied/will apply for a partial restriction of the digital copy of my thesis or dissertation.' Signed ……………………………………………........................... Date ……………………………………………........................... AUTHENTICITY STATEMENT ‘I certify that the Library deposit digital copy is a direct equivalent of the final officially approved version of my thesis. No emendation of content has occurred and if there are any minor variations in formatting, they are the result of the conversion to digital format.’ Signed ……………………………………………........................... Date ……………………………………………........................... 3 ABSTRACT T cell help to B cells is a fundamental property of adaptive immunity, yet only recently have many of the cellular and molecular mechanisms of T cell help emerged. T follicular helper (Tfh) cells are the CD4+ T helper cells that provide cognate help to B cells for high affinity antibody production in germinal centres (GC). This study has reveals a critical role of IL-21 in the upregulation of Tfh signature molecules. Expression of ICOS was found to be necessary for optimal production of IL-21; indicative of interplay between two Tfh expressed molecules. We also demonstrate that IL-21's costimulatory capacity for T helper differentiation operates at the level of the TCR through Vav1 signalling that controls T cell helper function and survival. Tfh cells express uniquely high levels of the IL-21 receptor relative to other T cell subsets, which reflects an IL-21- driven autocrine loop that is important for the generation and function of Tfh cells, which in turn were critical for supporting primary and secondary T dependent antibody responses. This study reveals a previously unappreciated role for Tfh cells in the formation of the GC through a CD4+ T cell intrinsic requirement for responsiveness to IL-21. IL-2 and IL-21 are crucial growth factors for distinct T helper subsets with opposing regulatory and effector functions, respectively. Mice made genetically deficient in IL-2 or its high affinity receptor chain (CD25) suffer from a fatal autoimmune disease characterized by ulcerative colitis and haemolytic anaemia. The observed autoimmunity and associated splenomegally are thought to be caused, in part, by a loss of regulation of effector T cells due to a deficit in IL-2- dependent Foxp3 regulatory T cells. Since IL-21 is known to facilitate the development of a number of autoimmune diseases, the possible contribution of IL-21 to the autoimmune pathology observed in Il2-/- mice was investigated in this study. Our findings demonstrate that IL-21:IL-21R signalling contributes to the destruction of tissues attributed to autoimmunity in Il2-/- mice and suggests that IL-21-producing T cells are significant targets of immune system regulation. 4 PUBLICATIONS, PRESENTATIONS AND PRIZES ARISING FROM THIS THESIS PUBLICATIONS Vogelzang, A., McGuire, H.M., Yu, D., Sprent, J., Mackay, C.R., and King, C. (2008). A fundamental role for interleukin-21 in the generation of T follicular helper cells. Immunity 29, 127- 137. Vogelzang, A., and King, C. (2008a). The modulatory capacity of interleukin-21 in the pathogenesis of autoimmune disease. Frontiers in Bioscience 13, 5304-5315. McGuire, H.M., Vogelzang, A., Hill, N., Flodstrom-Tullberg, M., Sprent, J., and King, C. (2009). Loss of parity between IL-2 and IL-21 in the NOD Idd3 locus. Proceedings of the National Academy of Sciences of the United States of America 106, 19438-19443. ORAL PRESENTATIONS A.Vogelzang, H. McGuire, J. Sprent, C. King. Interleukin-21 in a model of ulcerative colitis. Australasian Society for Immunology, NSW branch meeting. Bowral, Australia (September 2009) A.Vogelzang, H. McGuire, J. Sprent, C. King. The role of Interleukin-21 in T Follicular Helper cell differentiation. Australasian Society for Immunology, 37th Annual Scientific Meeting. Canberra, Australia (Dec 2008) A.Vogelzang, H. McGuire, J. Sprent, C. King. A fundemental role for Interleukin-21 in the generation of T Follicular Helper Cells. The Federation of Immunological Societies of Asia- Oceania 4th Congress. Taipei, Taiwan (October 2008) A.Vogelzang, H. McGuire, C. Mackay, J. Sprent, C. King. The role of Interleukin-21 in T Follicular Helper cell differentiation. Australasian Society for Immunology, 36th Annual Scientific Meeting. Sydney, Australia (Dec 2007) A.Vogelzang, H. McGuire, C. Mackay, J. Sprent, C. King. The role of Interleukin-21 in T Follicular Helper cell differentiation. 17th St Vincents & Mater Health Sydney Research Symposium. Sydney, Australia (Sept 2007) 5 POSTER PRESENTATIONS A.Vogelzang, H. McGuire, J. Sprent, C. King. A fundamental role for Interleukin-21 in germinal centre antibody responses. Immunology 2009 96th Annual Meeting with the American Association of Immunologists. Seattle WA, USA (May 2009). A.Vogelzang, H. McGuire, C. Mackay, J. Sprent, C. King. The role of Interleukin-21 in T Follicular Helper cell differentiation. Frontiers in Immunological Memory Newport Beach CA, USA (Septemeber 2008). PRIZES Postgraduate Research Student Support Scheme Award (UNSW) for travel to AAI Immunology 09 96th Annual Meeting with the American Association of Immunologists, Seattle, USA. 2009 Prestigious New Investigator Prize at the Australasian Society of Immunologists (ASI) Annual meeting, 2008. Australasian Society of Immunologists Travel Bursary of $3000 awarded to travel to the Federation of Immunological Societies of Asia-Oceania 4th Congress, 2008. 6 ACKNOWLEDGMENTS Thank you to my supervisor Cecile King for your guidance and constant enthusiasm during this project. Thanks also to Helen McGuire who has helped on so many long days in the laboratory, becoming a great friend in the process. I’ve really enjoyed working closely with you both, I think we make a great team and I look forward to working with you in the future. Thank you to Kylie Webster, Rachael Kohler, Jaeho Cho and Heeok Kim, Marcel Batten and Jon Sprent for all your sage advice during lab meetings and meetings in labs. Thank you to Chris and Yovina for running a wonderful flow cytometry service, the many BTF staff who have helped me in the past four years, and the staff of the Garvan Institute which has been a great workplace. Thank you also to the members of the Brink, Silveira, Tangey, 2xMackay and Sprent labs for your generous provision of reagents and help during my PhD. Lastly, thanks to all my friends and family for providing me with support, sympathy, and big dinners while I’ve been a student, especially Blake and my parents who were extremely generous with all of the above throughout. Experiments that were not the sole work of the author All experiments were performed by the author at the Garvan Institute, with the following exceptions; TaqMan ABI assays were carried out by Helen McGuire on samples prepared by the author; Paraffin embedding of histology samples was done by the St Vincents Hospital pathology unit, while sections and H&E staining of histology was performed by Alice Boulghourjian of the Garvan Institute Histology Unit. Assessment of pathology of the colon and pancreas sections was generously undertaken by Doctor Peter Earls of the St Vincents Hospital pathology unit. 7 TABLE OF CONTENTS 1 Introduction ................................................................................................................................... 18 1.1 A brief review of the mammalian immune system.............................................................. 18 1.1.1 Primary lymphoid organs.............................................................................................
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