Gosaccharides (Gos) Supplementation: a Randomized, Double-Blind Controlled Study in Healthy Aged Adults
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IMMUNOLOGICAL AND GASTROINTESTINAL EFFECTS OF GALACTOOLIGOSACCHARIDES (GOS) SUPPLEMENTATION: A RANDOMIZED, DOUBLE-BLIND CONTROLLED STUDY IN HEALTHY AGED ADULTS By STEPHANIE-ANNE GIRARD A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY UNIVERSITY OF FLORIDA 2012 1 © 2012 Stephanie-Anne Girard 2 To my parents, Yves and Danielle, and to all of those that have supported me during this incredible and life changing experience 3 ACKNOWLEDGMENTS Firstly, I would like to thank my mentor Dr. Bobbi Langkamp-Henken who has successfully guided me through this journey where many obstacles were encountered but all surpassed. She was always there to motivate and encourage me whilst helping me acquire new skills along the way. I would also like to acknowledge all of the members of my committee; Dr. Wendy Dahl, Dr. Susan Percival and Dr. Volker Mai who have challenged me and helped me attain the level of knowledge that I possess today. I give my thanks to my parents, Yves and Danielle, who have been with me, although not physically but rather emotionally and mentally, and without their support I surely could not have completed this degree. They have encouraged me throughout these three years and helped me reach my ultimate goal. I would certainly not have completed my PhD without the continual support from my lifelong friend and companion Kaunteya Nundy. I have to share this doctoral degree with him as we have together achieved this huge success. As we prepare to build a life together, I feel that we have accomplished this steppingstone to our prosperous, both with love and success, future together. A special thanks go to all of the current and former members of my laboratory; CJ Nieves, Gretchen Specht, Sam Spaiser, Ally Radford and Christine Hughes. They certainly made my days sunnier and kept my hopes up during hard times. I must also acknowledge all of the undergraduate students that have worked with me and helped me execute all of the laboratory assays as well as conduct this incredible study; Elaine Tan, Christina Vitale, Nicholas Morrell, Vivek Iyer, and Paula Beers. Additionally, fellow graduate students of collaborating laboratories such as Rebecca Creasy and Tyler Culpepper have always been there for me when I needed a 4 friend. They have helped me understand concepts and master techniques and without their support and friendship, I could not have excelled and achieved my goals. 5 TABLE OF CONTENTS page ACKNOWLEDGMENTS .................................................................................................. 4 LIST OF TABLES ............................................................................................................ 9 LIST OF FIGURES ........................................................................................................ 10 LIST OF ABBREVIATIONS ........................................................................................... 11 ABSTRACT ................................................................................................................... 14 CHAPTER 1 INTRODUCTION AND SPECIFIC AIMS ................................................................. 16 2 REVIEW OF THE LITERATURE ............................................................................ 18 Introductory Notes .................................................................................................. 18 Galactooligosaccharides ......................................................................................... 19 Properties ......................................................................................................... 19 GRAS Status and Health Claims ...................................................................... 20 Research Involving Galactooligosaccharides ................................................... 21 Stool improvement ..................................................................................... 23 Mineral absorption ..................................................................................... 24 Lipid metabolism ........................................................................................ 25 Immunomodulation .................................................................................... 26 Carcinogenesis .......................................................................................... 28 Common Cold Symptoms and Complementary Health Practices ........................... 28 Cold Assessment in Research ......................................................................... 30 Nutritional Status, Immune Function and Common Cold .................................. 31 Nutritional Modulation of Common Cold ........................................................... 33 Echinacea .................................................................................................. 34 Ascorbic acid .............................................................................................. 35 Zinc ............................................................................................................ 36 Multivitamins and minerals ......................................................................... 39 Garlic ......................................................................................................... 39 Probiotics ................................................................................................... 41 Prebiotics ................................................................................................... 42 Age and Common Cold .................................................................................... 43 Immune System ...................................................................................................... 45 Leukocytes ....................................................................................................... 45 Serum Acute Phase Proteins ........................................................................... 46 Peripheral Blood Mononuclear Cell (PBMC) Cytokine Production to Mitogens........................................................................................................ 47 Immunosenescence ......................................................................................... 50 6 Mucosal Immunity ............................................................................................ 52 Salivary and Fecal Secretory IgA (sIgA) ........................................................... 54 Digestive Health ...................................................................................................... 56 Microbiota ......................................................................................................... 56 Gastrointestinal Symptom Rating Scale ........................................................... 57 GOS and the Microbiota ................................................................................... 58 Aged Adults and the Microbiota ........................................................................ 59 3 METHODS .............................................................................................................. 66 Participants ............................................................................................................. 66 Experimental Design ............................................................................................... 67 GOS Administration Protocol .................................................................................. 68 Influenza Vaccination .............................................................................................. 69 Study Questionnaires.............................................................................................. 69 Mini Nutritional Assessment .................................................................................... 71 Food Frequency Questionnaire (FFQ) .................................................................... 73 Reagents ................................................................................................................ 74 Blood Sample Collection ......................................................................................... 74 Serum Collection .................................................................................................... 75 Phenotypic Determination of Lymphocyte Population ............................................. 75 CRP Assay ............................................................................................................. 77 PBMC Isolation ....................................................................................................... 77 PBMC Stimulation with Mitogens (PHA and LPS) Assay ........................................ 78 Multiplex Bead Based Assay .................................................................................. 78 Saliva Collection ..................................................................................................... 79 Salivary sIgA Assay ................................................................................................ 80 Fecal Collection ...................................................................................................... 80 Fecal sIgA Assay .................................................................................................... 81 Microbiota Analysis ................................................................................................