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Conceptus-Derived Immunosuppressive Factors and Their Role in Fetal Allograft Rp Otection

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Conceptus-Derived Immunosuppressive Factors and Their Role in Fetal Allograft Rp Otection Louisiana State University LSU Digital Commons LSU Historical Dissertations and Theses Graduate School 1991 Conceptus-Derived Immunosuppressive Factors and Their Role in Fetal Allograft rP otection. Terri Lynn Roth Louisiana State University and Agricultural & Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_disstheses Recommended Citation Roth, Terri Lynn, "Conceptus-Derived Immunosuppressive Factors and Their Role in Fetal Allograft rP otection." (1991). LSU Historical Dissertations and Theses. 5207. https://digitalcommons.lsu.edu/gradschool_disstheses/5207 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Historical Dissertations and Theses by an authorized administrator of LSU Digital Commons. For more information, please contact [email protected]. INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand corner and continuing from left to right in equal sections with small overlaps. Each original is also photographed in one exposure and is included in reduced form at the back of the book. Photographs included in the original manuscript have been reproduced xerographically in this copy. Higher quality 6" x 9" black and white photographic prints are available for any photographs or illustrations appearing in this copy for an additional charge. Contact UMI directly to order. University Microfilms International A Beil & Howell Information Com pany 300 North Zeeb Road, Ann Arbor, Ml 48106-1346 USA 313/761-4700 800/521-0600 Order Number 9207529 Conceptus-derived immunosuppressive factors and their role in fetal allograft protection Roth, Terri Lynn, Ph.D. The Louisiana State University and Agricultural and Mechanical Col., 1991 UMI 300 N. ZeebRd. Ann Arbor, MI 48106 CONCEPTUS-DERIVED IMMUNOSUPPRESSIVE FACTORS AND THEIR ROLE IN FETAL ALLOGRAFT PROTECTION A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Doctor of Philosophy in The Department of Animal Science by Terri Lynn Roth B.S., University of California Davis, 1986 M.S., University of California Davis, 1988 August 1991 ACKNOWLEDGEMENTS I would first like to thank ray major professor, Dr. Kenneth L. White, for initially convincing me to pursue iry Ph.D. degree at Louisiana State University, then for providing the opportunity for me to carry out the research in which I was most interested and finally, for his contributions to this research. I am also grateful to Dr. Donald L. Thorrpson, Jr. for kindly allowing me to use his horses. My sincere appreciation is extended to Dr. David W. Horohov, both for his helpful advice and for sharing with me his extensive knowledge of immunology. My gratitude is also extended to Dr. Kenneth L. White, Dr. David W. Horohov, Dr. Donald L. Thatpson, Jr., Dr. Lee L. Southern, Dr. William Hansel and Dr. Joseph C. Newton for their helpful suggestions in preparing this dissertation and for serving as members of iry committee. I would like to thank my colleagues, Saeed Rahmanian, Don Colbom, Joe Capehart, Brett Barry and Mary Rabb for their assistance at the horse farm. Special thanks is extended to Dr. Bill Swanson for including me in his work at the local zoos and for offering his invaluable friendship. Finally, I would like to thank the L.S.U. Board of Regents for the generous fellowship that supported me during this doctoral program. TABLE OF CONTENTS PAGE AOONCWLEDGEMENTS ........................................ ii LIST OF TABLES .......................................... vi LIST OF FIGURES ........................................ vii LIST OF ABBREVIATIONS ................................. viii ABSTRACT ................................................ ix INTRODUCTION ............................................ xi CHAPTER I. Literature Review............................... 1 IMMUNE RESPONSES AND ALLOGRAFT REJECTION.................. 1 The History of Allografts............................ 1 Humoral Irrenune-mediated Graft Rejection................ 2 Cell-mediated Graft Rejection......................... 3 T Lymphocytes „ ........................... 3 T Lymphocyte Activation............................ 5 The IL-2 Receptor................................. 5 Cytotoxic T Lymphocytes............................ 6 Additional Cell Types Affected by IL-2............... 7 Suppressor Cells.................................. 8 In Vitro Assessment of Immune Cell Function............. 9 Mitogen-stimulated Proliferation.................... 9 Mixed Lymphocyte Reaction. ...................... 9 Cytolytic Cell Assays............................ 10 THE PUTATIVE IMMUNO-PROTECTIVE ROLE OF THE PLACENTA......... 11 The Phenomenon of the Fetal Allograft................. 11 The Equine Placenta................................. 12 Hybrid and Interspecific Pregnancies.................. 14 The Murine Model................................. 15 The Sheep/Goat Model............................. 17 The Equine Model................................. 19 Exotic Interspecies Pregnancies.................... 21 Embryo Manipulation................................. 23 IMMUNOSUPPRESSION AND PREGNANCY......................... 26 Antibody Involvement................................ 26 iii Suppressor Cells................................... 27 Macrophage Involvement in Immunosuppression............ 28 Suppressor Activity of Prostaglandins and Hormones....... 28 Soluble Suppressor Factors........................... 30 Murine......................................... 30 Human.......................................... 32 tftigulate....................................... 33 Rationale of Present Research........................ 35 CHAPTER II. Suppression of Sheep and Goat lyirphocyte proliferation fcy sheep, goat and sheep x goat hybrid trophoblast tissue cultures............... 38 HSFTROEOSTION......................................... 38 MATERIALS AND METHODS.................................. 39 RESULTS.............................................. 42 DISCUSSION........................................... 45 CHAPTER III. Suppression of lyirphocyte proliferation fcy a >30,000 MW factor in horse conceptus- conditioned medium............................ 52 INTRODUCTION......................................... 52 MATERIALS AND METHODS.................... 54 RESULTS.............................................. 58 DISCUSSION........................................... 66 CHAPTER IV. PGE2 - Independent immunosuppressive activity of horse trophoblast tissue..................... 72 INTRODUCTION......................................... 72 MATERIALS AND METHODS.................................. 73 RESULTS.............................................. 75 DISCUSSION........................................... 77 CHAPTER V. Horse conceptus-derived suppression of T and B lymphocyte proliferation: interleukin-2 receptor involvement............................ 82 INTRODUCTION......................................... 82 MATERIALS AND METHODS.................................. 85 RESULTS.......... 96 iv DISCUSSION......... 107 SUMMARY AND CONCLUSIONS.................................. 118 LITERATURE CITED ........................................ 123 A P P E N D I X ............................................... 148 VITA .................................................... 151 v LIST OF TABLES TABLE PAGE 1. Suppression of FWM-stimulated sheep and goat lymphocyte proliferation fcy caprine, hybrid and ovine trophoblast tissue-conditioned medium................. 46 2. Effect of control medium and HCCM from Groups I-III on mitogen-stimulated horse PBL proliferation...........59 3. Effect of treated HCCM on FWM-stimulated horse PBL proliferation................................... 61 4. PGE2 concentrations in horse trophoblast- conditioned medium (HTCM) and indomethacin- treated HTCM (I-HTCM).................................... 78 5. The effect of HCCM I and HCCM II on FWM- stimulated lynphocytes................................... 87 6. Effect of HCCM II supplementation to mitogen- stimulated lynphocytes............ 97 7. Effect of fractionated HCCM II and CM on CTLL-2 cells and murine splenocytes............................. 105 8. Fluorescence activated cell sorter analysis................. 106 vi LIST OF FIGURES FIGURE PAGE 1. The effect of caprine, ovine and hybrid trophoblast tissue-conditioned medium on FWM-stimulated sheep lyirphocyte proliferation................................. 43 2. The effect of caprine, ovine and hybrid trophoblast tissue-conditioned medium on FWM-stimulated goat lyirphocyte proliferation................................
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