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The Impact of Diet on Immunosenescence

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The Impact of Diet on Immunosenescence The Impact of Diet on Immunosenescence Sarah Jayne Clements, BSc (Hons) A thesis submitted in accordance with the requirements for the degree of Doctor of Philosophy (PhD) To The University of East Anglia Institute of Food Research Gut Health & Food Safety Norwich Research Park Colney Lane Norwich NR4 7UA April 2017 This copy of the thesis has been supplied on condition that anyone who consults it is understood to recognise that its copyright rests with the author and that use of any information derived there from must be in accordance with current UK Copyright Law. In addition, any quotation or extract must include full attribution. 1 Abstract Introduction: The population is ageing but this accompanies increased susceptibility to infection and age-associated diseases, as well as reduced vaccination responses; potentially attributable to reduced immune function. Immunosenescence describes the deleterious effects of ageing on the immune system and is associated with a chronic, low-grade, inflammatory state; inflammaging. Habitants of Mediterranean regions maintain good health into old age; often attributed to Mediterranean (MED)- diets. Hypothesis: Adoption of a MED-diet by elderly subjects, in Norfolk, may improve immune responses of these individuals; particularly in terms of dendritic cell (DC) function and antibody diversity. Experimental approach: Elderly subjects recruited onto the Nu-AGE study were randomised to the control or MED-diet groups, for one year. Blood samples were compared from pre- and post-intervention, and to blood samples from young subjects. Study compliance was assessed using high performance liquid chromatography-with tandem mass spectrometry (HPLC-MS/MS) analysis of urine samples. Immune cell subset numbers and concentrations of secreted proteins were determined by flow cytometry, after staining for surface markers and intracellular proteins. Age and dietary impact on antibody diversity was quantitated using a novel-polymerase chain reaction (PCR)-based technique developed by the Babraham Institute. Results: The MED-diet group had higher urinary hydroxytyrosol sulphate post- intervention but self-reported diet diary analyses showed no difference in MED-diet scores. Reduced myeloid DC numbers were observed in blood samples from elderly subjects compared to young. The elevated secretion of the adipokine, resistin, after ex vivo stimulation of peripheral blood mononuclear cells (PBMCs) from elderly subjects, was significantly reduced after MED-diet intervention, but this change from baseline was not significantly different to the control group. Antibody diversity was reduced with age, dietary intervention may prevent further reductions in unique clonotypes. Conclusions: Further evidence of numerical and functional effects of ageing on DCs, are shown. The MED-diet showed potential to impact on the ageing immune cells investigated and could provide an economical approach to address problems associated with our ageing population. 2 Acknowledgements This PhD was supported by a three-year studentship provided by Norwich Medical School at the University of East Anglia. I would first like to express my deepest thanks to my primary supervisor, Professor Simon Carding, for the extensive support and guidance you have provided throughout my PhD and to my secondary supervisor, Professor Claudio Nicoletti, for many helpful conversions during the method development stages. You have both been a tremendous support over the years. I would also like to thank all members of the Carding lab group, past and present, who have been such a great group of people to work alongside and learn from, you have provided some wonderful memories and valuable lessons along the way. With particular thanks to Dr Isabelle Hautefort, Dr Emily Jones, Dr Ana Carvalho, Dr Elizabeth Bassity and Dr Jo Brooks, you have not just been colleagues but true friends who have made my time at IFR even more enjoyable. I would particularly like to thank Dr Monica Maijo Ferre for your help and friendship while working together on the Nu-AGE study, along with Dr Kamal Ivory and Dr Amy Jennings. Also, to Sally Bailey and your team for allowing me into your phlebotomy department. With particular thanks to all study volunteers for agreeing to donate blood for my research. To my collaborators at the Babraham Institute, Dr Anne Corcoran, Dr Daniel Bolland, Dr Louise Mattheson and Peter Chovanec, thank you for introducing me to the fascinating world of VDJ recombination and for the many hours of your time you have dedicated to the experimental work and analysis. Also, thank you to Mark Philo and Dr Gwen Le Gall for the hard work in conducting the metabolomics analysis of my samples. I would also like to thank my examiners, Professor Ian Clark and Professor Christine Williams, for kindly agreeing to discuss my work with me. I have made many friends while at IFR and I would like thank you all for the interesting conversations, the fantastic Christmas parties (and outstanding fancy dress), the brilliant student lunches and the many kind words over the years. Rebecca Edwards, Lizzie Thursby, Alistair Walsham and Dan Lock thank you for being the best group of students to go through this journey with, we have some fantastic memories from over the years. I would not have finished my PhD had it not been for two incredibly dedicated medics. Dr Karanth, you have looked out for me from the outset and your care is always 3 second to none, I thank you from the bottom of my heart for everything you have done for me. Mr Khan, you literally saved my life on the operating table. You both are the reason I managed to finish my PhD, I cannot thank you enough. To my wonderful friends, Dr Harri Bell, Dr Becca Lancaster, Jess Lefley, Emma Nicholls, Becky Carlton, Alex Bennett, Tom Gibbard, Frann Goodge and Adam Goodge thank you for providing me with much needed diversions from my studies, for the fantastic holidays and the much needed words of support. Finally, and most importantly, I have the most supportive, loving and understanding family. Mum and Dad thank you for everything that you have done for me over the years, without your support I could never have started my PhD, let alone finish it. You have always known what to do or say during the ups and downs of my PhD and have always believed in me, I’m just sorry for my moods, tears and absence. Rachel and James thank you for letting me come over to stay so many times, for distracting me when I needed a break and for always being there when I needed a chat. Chris you have been my rock. Thank you for understanding and just being there, for all of the little things that you’ve done and for the many laughs along the way. You have all ensured I got to this point and have never stopped believing in me. I dedicate this thesis to my late Nan, Kath Kirk, the woman I always looked up to and strive to be like; I hope you would be very proud of me. 4 Table of Contents Abstract .................................................................................................................. 2 Acknowledgements ............................................................................................... 3 Table of Contents .................................................................................................. 5 List of Figures ...................................................................................................... 12 List of Tables ....................................................................................................... 15 Abbreviations ....................................................................................................... 15 Chapter 1 .............................................................................................................. 25 1.1.1 Ageing .............................................................................................. 25 Impact of ageing on physiology and organ function .......................... 25 Body composition ........................................................................................ 26 Gastrointestinal function .............................................................................. 27 Immunological function................................................................................ 28 Ageing of innate immune cells .......................................................... 33 Monocytes/ macrophages ........................................................................... 33 Neutrophils .................................................................................................. 34 Natural Killer (NK) cells ............................................................................... 35 Dendritic cells (DCs) ................................................................................... 36 Ageing of primary and secondary lymphoid organs .......................... 39 Ageing of adaptive immune cells ...................................................... 40 Humoral ...................................................................................................... 41 Cell mediated .............................................................................................. 43 Leukocyte trafficking between the gut and the bloodstream .............. 46 Microbiota and its effect on the immune system ............................... 47 Diet ................................................................................................... 50 Carbohydrates ...........................................................................................
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