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Phd Thesis – Title Page Nayyar Ahmed PhD Thesis – Title Page Nayyar Ahmed Identification and Characterisation of a Novel Interleukin-4 Receptor Antagonist for Allergy Treatment ___________________ by Nayyar Ahmed B.Biomed.Sc. (Honours) Submitted in fulfilment of the requirements for the degree of Doctor of Philosophy Deakin University February, 2015 i PhD Thesis – Acknowledgements Nayyar Ahmed Acknowledgements Firstly, I would like to thank Associate Professor Cenk Suphioglu for being such a caring and supportive person throughout my PhD. Your friendliness surpasses your duty as a professor and no matter how many words I spill on this paper, they will never be enough. Thank you for being so patient with my work. Thank you once again, for all your efforts and constant guidance throughout my candidature. I am also grateful to you for giving me the opportunity to be a part of the NARL laboratory. I wasn’t sure if I wanted to pursue a career as a research scientist, however after joining your laboratory, you have inspired me and increased my passion for scientific research. Ever grateful to you! I would like to thank my co-supervisors Professor Colin Barrow and Professor Leigh Ackland for serving as my co-supervisors for my candidature. I also want to thank you for your brilliant comments and suggestions. I would also like to thank Professor Andrew Sinclair and Dr. Philip Taylor for their unprecedented support for my research. My sincere thanks to Nadia Sadli, who helped me with tissue culture work. Thanks Nadia for being such a caring friend and a wonderful lab mate. And thanks to all the members of NARL, God Bless you all! My very special thanks to Pathum (MEYA). You have been like a brother to me, thanks a lot for making my candidature such a joy. I would be privileged, if I ever get the opportunity to work with you again! I’ll miss the good times we’ve spend together. Thanks for all the favours you have done for me. You are nothing less than an angel. iv PhD Thesis – Acknowledgements Nayyar Ahmed My sincere gratitude goes to the Deakin University security guards, Tom, Dayle, Alex, Rohit and Shiv. Thanks for all the good times and late-night chats. Thank you for making me feel safe while working in the lab around the clock. I am ever grateful to you. I would like to express my appreciation to my beloved wife Mashal Pall who spent sleepless nights with me and was always my support in moments when there was no one to answer my queries. Thank you for your timely support and your words of encouragement. I would like to extend my sincere and heart-filled thanks to my wife’s parents (Waseem Pall & Farhat Pall) and siblings (Mehwish Pall, Sabahat Pall and Zoha Pall). One can only wish for such wonderful in-laws like yourselves. Thanks for making my wedding a success and for all the kind favours you did for me. I am forever grateful to you all. Last but not the least, I want to extend my gratitude and love to my family members, my father Shams Daud (Abbu), my beloved mother Attiya Shams (Ammi), my brother Nasir Ahmed and his wife Ajam Ahmed. Thank you Abbu & Ammi for your encouraging words and unconditional love for which I am ever thankful and indebted to you. Your prayer for me was what sustained me thus far. Thanks for your emotional support; you guys have stood by me in my times of weakness. And thank you for waiting so patiently for me to finish my studies. I have been away from you both far too long. Finally I thank my God Almighty for letting me through all the difficulties. I have experienced Your guidance day by day. You are the one who let me finish my degree. I will keep on trusting You for my future. Thank you, Lord. v PhD Thesis – List of Publications Nayyar Ahmed List of Publications Journal Articles: Ahmed N, Dhanapala P, Suphioglu C. Identification and characterization of a novel IL-4 receptor α chain (IL-4Rα) antagonist to inhibit IL-4 signalling. Cell Physiol Biochem 2015; 36:831-842 (Chapter 2 & 3 results published) Ahmed N, Dhanapala P, Sadli N, Barrow CJ, & Suphioglu C. Mimtags: The use of phage displaytechnology to produce novel protein-specific probes. Journal of Immunological Methods. 2014; 405(0):121-9 (Chapter 2 results published). Ahmed N, Barrow CJ, Sinclair A, Suphioglu C. Discovering the effects of omega-3 and omega-6 fatty acids on allergy using a HEK-Blue cell-line. International Journal of Molecular Sciences (Chapter 4 results submitted for publication). Ahmed N, Barrow CJ, Adcock J & Suphioglu C. Discovering the effects of CoQ10 and fatty acid derived resolvins on allergy using a HEK-Blue cell-line. International Journal of Molecular Sciences (Chapter 5 results under preparation). Taylor PE, Ahmed N, Suphioglu C. Relationship between ryegrass pollen count and weather variables in regional and urban Victoria, Australia. (Chapter 6 results under preparation) Book Chapters: Ahmed N, Dhanapala P, Suphioglu C., (2014). Nothing to be sneezed at: Th2 cytokines as novel therapeutic targets for the production of anti-allergy agents, Bentham Science Publishers (eds), frontiers in clinical drug research anti-allergy agents (Chapter 1 literature review submitted). Sadli N, Ahmed N, Ackland M.L, Sinclair A, Barrow C & Suphlioglu, C., (2011). Effect of zinc and DHA on expression levels and post-translational modifications of histones H3 and H4 in human neuronal cells, in Raymond, Chuen-Chung Chang (eds), Neurodegenerative diseases - processes, prevention, protection and monitoring, pp. 141-164, InTech, Rijecka, Croatia. Conference Papers (abstracts published): Presented at the European Academy of Allergy and Clinical Immunology (EAACI) conference Ahmed N, Dhanapala P, Sadli N, Barrow CJ, & Suphioglu C, 2014, ‘Mimtags: the use of phage display technology to produce novel protein specific probes’, Allergy, vol. 69, p. 298. vi PhD Thesis – List of Publications Nayyar Ahmed Ahmed N & Suphioglu C, 2014, ‘Discovering the effects of omega-6 fatty acids on allergy using a HEK-Blue cell line’, Allergy, vol. 69, p. 534. Ahmed N, Ramsbottom K, Carr S & Suphioglu C, 2013, ‘Identification and characterization of IL-4 and IL-4 receptor as novel intervention targets for allergy treatment’, Allergy, vol. 68, p. 470. Ahmed N, Barrow CJ, Adcock J, Sinclair A & Suphioglu C, 2013, ‘Importance of CoQ10 and omega-3 fatty acids as novel therapeutics for allergy’, Allergy, vol. 68, p. 344. Ahmed N, Dhanapala P, Carr S & Suphioglu C, 2011, 'Characterisation of a novel IL-4 receptor antagonist for allergy treatment', Allergy, vol. 66, p. 509-10. Presented at the Australasian Society for Clinical Immunology and Allergy (ASCIA) conference Ahmed N & Suphioglu C, 2014, ‘Discovering the effects of omega-3 and omega-6 fatty acids on allergy using a HEK-Blue cell line’, Internal Medicine Journal, vol. 44, p. 1. Ahmed N, Dhanapala P & Suphioglu C, 2013, 'Discovering the effects of Omega-3 fatty acids on allergy using a HEK-Blue cell line', Internal Medicine Journal, vol. 43, p. 1. Ahmed N, Dhanapala P & Suphioglu C, 2013, 'Identification and characterisation of a novel cytokine IL-13 antagonist for allergy treatment', Internal Medicine Journal, vol. 43, p. 1. Ahmed N, Dhanapala P & Suphioglu C, 2012, 'Identification and characterization of a novel IL-4 receptor antagonist for allergy treatment', Internal Medicine Journal, vol. 42, p. 1. vii PhD Thesis – Table of Contents Nayyar Ahmed Tables of Contents Page number Title page……………………………………………………………………….i Deakin University – ACCESS TO THESIS- A………………………………..ii Candidate Declaration Form…………………………………………………...iii Acknowledgements…………………………………………………………....iv List of Publications…………………………………………………………….v Table of Contents……………………………………………………………...viii Abstract………………………………………………………………………..xviii List of Abbreviations…………………………………………………………..xxi Chapter 1 – Literature review………………………………………………….1 1.1 Introduction of Allergy and Allergens…………………………………...2 1.1.1 Sources of Allergens………………………………………………….2 1.1.2 Symptoms of Allergy…………………………………………………4 1.1.3 Rising Incidence/Trends in Allergic Disease…………………………4 1.2 Overview of Allergy as a Disease: Atopy in Allergic Individuals………5 1.2.1 Underlying Causes of Allergy in Atopic individuals…………………5 1.2.2 Diagnostic Tests for Allergic Conditions……………………………..9 1.2.2.1 RAST…………………………………………………………....9 1.2.2.2 Skin Prick Tests…………………………………………………9 1.2.2.3 Blood Specific IgE Testing……………………………………..9 1.2.3 Current Available Treatments………………………………………..10 1.3 Immune Response: Allergic vs. Non-Allergic Response………………..11 1.3.1 Non-Allergic Immune Response……………………………………..11 viii PhD Thesis – Table of Contents Nayyar Ahmed 1.3.2 Allergic Immune Response…………………………………………..12 1.3.2.1 Humoral Response in Allergic Individuals…………………….12 1.3.2.2 Cell-Mediated Response in Allergic Individuals…………………..14 1.4 Role of Cytokines in Immunity…………………………………………..14 1.4.1 Cytokines IL-4 and IL-13…………………………………………….15 1.4.2 Comparative Roles of IL-4/IL-13 with IL-4Rα chain………………..17 1.5 IL-4Rα as an Important Target for Allergy Treatment………………..19 1.5.1 Anti-IL-4R Monoclonal Antibody…………………………………...19 1.5.2 Soluble IL-4 Receptor (sIL-4R)……………………………………...21 1.5.3 Targeting IL-4R with Hybrid Peptides………………………………21 1.5.4 Further Analysis of Blocking IL-4R………………………………....22 1.5.5 Current Available Treatments for Targeting IL-4 and IL-13 Cytokines……………………………………………………….,22 1.5.5.1 Specific Immunotherapy (SIT)…………………………………23 1.5.5.2 Modified Recombinant Vaccines…………………………….…25 1.5.5.3 Allergoids………………………………………………….……25 1.5.5.4 Adjuvants……………………………………………………….26 1.5.5.5 Peptide Immunotherapy………………………………………...26 1.5.5.6 Plasmid Deoxyribonucleic
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