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Characterizing Humidity, Sex, and B-Cell Gene Regulation in Fungal Allergic

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Characterizing Humidity, Sex, and B-Cell Gene Regulation in Fungal Allergic CHARACTERIZING HUMIDITY, SEX, AND B-CELL GENE REGULATION IN FUNGAL ALLERGIC ASTHMA A Thesis Submitted to the Graduate Faculty of the North Dakota State University of Agriculture and Applied Science By Emma Claire Kusick In Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE Major Department: Microbiological Sciences April 2020 Fargo, North Dakota North Dakota State University Graduate School Title CHARACTERIZING HUMIDITY, SEX, AND B-CELL GENE REGULATION IN FUNGAL ALLERGIC ASTHMA By EMMA KUSICK The Supervisory Committee certifies that this disquisition complies with North Dakota State University’s regulations and meets the accepted standards for the degree of MASTER OF SCIENCE SUPERVISORY COMMITTEE: Dr. Jane Schuh Chair Dr. Teresa Bergholz Dr. Penelope Gibbs Dr. Sumit Ghosh Approved: 5/4/2020 Dr. John McEvoy Date Department Chair ABSTRACT Asthma is a debilitating lung disease that affects nearly 300 million people worldwide. Environments with high humidity and subsequent mold exposure often trigger allergic asthma. Sex differences have been reported in the incidence, prevalence, and severity of asthma. B-lymphocytes are recruited in high numbers to the allergic lung in response to the inhalation of Aspergillus fumigatus mold spores (conidia). In this work, we used a mouse model of allergic fungal asthma to assess environmental humidity, sex, and B-lymphocytes in an inhalational model of allergic fungal asthma. Our results showed that animals sensitized in low humidity conditions had no airway hyperresponsiveness (AHR), inflammation, but an increase in IgG3 antibody production. Males weighed more than females, female mice had more fibrosis and produced more IgG3 Ab, but sex showed no impact on low humidity. C19+ B- lymphocytes differentially downregulated multiple genes related to allergic asthma returning the body to homeostasis. iii ACKNOWLEDGEMENTS Earning my M.S. has been one of the most challenging, but exciting years of my life. The two additional years spent at NDSU were well worth the growing pains, and I gained more friendships and learned so much about myself to be successful in life. First, I would like to thank Dr. Jane Schuh for being my adviser during this experience. She was always there to provide guidance, direction, and support throughout the process. I have learned so much about how to become a good scientist and solve problems in different ways. You are truly a strong female role model of mine, and I appreciate that you were willing to take me under your wing. Thank you for refusing to let me give up when times were rough and showed me that this degree is for myself. Second, I would like to thank Scott Hoselton for being the mentor, teacher, listener, advice giver, and so on. You answered question after question, and without you this would not have been possible. You are the person holding the Schuh laboratory steady, and you have helped me become a better- trained researcher. Third, I would like to thank my committee members, Dr. Teresa Bergholz, Dr. Penelope Gibbs, and Dr. Sumit Ghosh for their support and helpful suggestions. Although our time was brief together, I enjoyed the committee meetings and you made a learning and encouraging environment. Dr. Bergholz, I appreciated that your door was always open, and I enjoyed working with you during my time as the Graduate Student Association President. Dr. Gibbs, I could always find a smile from you and enjoyed hearing all your stories about life. Dr. Ghosh, without you and your previous research, my research would not exist. You were amazing to learn from as both an undergraduate and graduate student. To the members of the Schuh lab over the years, you have been a source of friendship, encouragement, and advice in the classroom, laboratory, and life throughout these years. I could not have made it through all this without you- Thank you! Friends and Family: I have had the pleasure of gaining many friends throughout my undergraduate and graduate career at NDSU. Although many of them left after undergrad, they were still there to support during this journey. I was able to meet new graduate students from other departments that soon became friends and iv fellow fighters through all this. Thank you to everyone that told me I could do this, and I hope to return the support and laughter someday. I could not have achieved this without my parents, Bo and Cheryl Kusick, and my brother Owen Kusick. You have shown me that dedication, hard work, and encouragement will allow me to follow my dreams and that you will always be there to help pick me up if I falter. The four of us can get through anything, and I owe you three everything. Finally, I wish to thank my husband, Kyle Mason, for everything he has done for me and supporting me while I finished up my degree. Having you there to listen about frustrating days when everything had gone wrong even though you did not understand exactly what I was doing helped me immensely. I am finally finished and thanks for waiting for me so we can begin our next adventure. v DEDICATION I dedicate this to all the strong women in my life who taught me to be fearless. Family, friends, mentors, and colleagues who showed me that a girl can do anything she sets her mind to. vi TABLE OF CONTENTS ABSTRACT .............................................................................................................................................iii ACKNOWLEDGEMENTS ....................................................................................................................... iv DEDICATION ......................................................................................................................................... vi LIST OF TABLES ................................................................................................................................... ix LIST OF FIGURES .................................................................................................................................. x LIST OF ABBREVIATIONS .................................................................................................................... xii 1. LITERATURE REVIEW ....................................................................................................................... 1 1.1. Asthma ......................................................................................................................................... 1 1.2. Model............................................................................................................................................ 4 1.3. Immune Response to Fungus ....................................................................................................... 7 1.4. Sex Differences............................................................................................................................. 9 1.5. B-lymphocytes ............................................................................................................................ 10 1.6. Summary .................................................................................................................................... 13 2. CHARACTERIZATION OF FUNGAL ALLERGIC ASTHMA IN DRY CONDITIONS ............................ 15 2.1. Introduction ................................................................................................................................. 15 2.2. Materials and Methods ................................................................................................................ 16 2.3. Results ....................................................................................................................................... 19 2.4. Discussion .................................................................................................................................. 25 3. CHARACTERIZATION OF SEX IN FUNGAL ALLERGIC ASTHMA IN DRY CONDITIONS ................ 29 3.1. Introduction ................................................................................................................................. 29 3.2. Materials and Methods ................................................................................................................ 29 3.3. Results ....................................................................................................................................... 32 3.4. Discussion .................................................................................................................................. 40 4. FUNGAL ALLERGIC B-LYMPHOCYTE GENE REGULATION ........................................................... 43 4.1. Introduction ................................................................................................................................. 43 4.2. Materials and Methods ................................................................................................................ 43 4.3. Results ....................................................................................................................................... 48 vii 4.4. Discussion .................................................................................................................................. 56 5. GENERAL DISCUSSION................................................................................................................... 64 6. CONCLUSIONS ................................................................................................................................ 69 REFERENCES
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