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Characterization of Immunity to Pasteurella Multocida Robert Morris Nathanson Iowa State University

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Characterization of Immunity to Pasteurella Multocida Robert Morris Nathanson Iowa State University Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 1981 Characterization of immunity to Pasteurella multocida Robert Morris Nathanson Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Allergy and Immunology Commons, Immunology and Infectious Disease Commons, and the Medical Immunology Commons Recommended Citation Nathanson, Robert Morris, "Characterization of immunity to Pasteurella multocida " (1981). Retrospective Theses and Dissertations. 6838. https://lib.dr.iastate.edu/rtd/6838 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. INFORMATION TO USERS This was produced from a copy of a document sent to us for microfilming. While the most advanced technological means to photograph and reproduce this document have been used, the quality is heavily dependent upon the quality of the material submitted. The following explanation of techniques is provided to help you understand markings or notations which may appear on this reproduction. 1. The sign or "target" for pages apparently lacking from the document photographed is "Missing Page(s)". If it was possible to obtain the missing page(s) or section, they are spliced into the film along with adjacent pages. This may have necessitated cutting through an image and duplicating adjacent pages to assure you of complete continuity. 2. When an image on the film is obliterated with a round black mark it is an indication that the film inspector noticed either blurred copy because of movement during exposure, or duplicate copy. Unless we meant to delete copyrighted materials that should not have been filmed, you will find a good image of the page in the adjacent frame. If copyrighted materials were deleted you will find a target note listing the pages in the adjacent frame. 3. When a map, drawing or chart, etc., is part of the material being photo­ graphed the photographer has followed a definite method in "sectioning" the material. It is customary to begin filming at the upper left hand corner of a large sheet and to continue from left to right in equal sections with small overlaps. If necessary, sectioning is continued again—beginning below the first row and continuing on until complete. 4. For any illustrations that cannot be reproduced satisfactorily by xerography, photographic prints can be purchased at additional cost and tipped into your xerographic copy. Requests can be made to our Dissertations Customer Services Department. 5. Some pages in any document may have indistinct print. In all cases we have filmed the best available copy. University Microfilms international 300 N. ZEEB RD., ANN ARBOR, Ml 48106 8122547 NATHANSON, ROBERT MORRIS CHARACTERIZATION OF IMMUNITY TO PASTEURELLA MULTOCIDA Iowa State University PH.D. 1981 University Microfilms InterriBtiOnsi 300 N. zeeb Road. Ann Arbor,MI 48106 PLEASE NOTE: In all cases this material has been filmed in the best possible way from the available copy. Problems encountered with this document have been identified here with a check mark V . 1. Glossy photographs or pages 2. Colored illustrations, paper or print 3. Photographs with dark background 4. Illustrations are poor copy 5. Pages with black marks, not original copy 6. Print shows through as there is text on both sides of page 7. Indistinct, broken or small print on several pages 8. Print exceeds margin requirements 9. Tightly bound copy with print lost in spine 10. Computer printout pages with indistinct print 11. Page(s) lacking when material received, and not available from school or author. 12. Page(s) seem to be missing in numbering only as text follows. 13. Two pages numbered . Text follows. 14. Curling and wrinkled pages 15. Other University Microfilms International Characterization of immunity to Pasteurella multocida by Robert Morris Nathanson A Dissertation Submitted to the Graduate Faculty in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY Major: Immunobiology Approved: Signature was redacted for privacy. In Charge of M, Signature was redacted for privacy. For the Major Department Signature was redacted for privacy. For the Graduate College Iowa State University Ames, Iowa 1981 ii TABLE OF CONTENTS Page INTRODUCTION 1 LITERATURE REVIEW 4 PART I: PASTEURELLA MULTOCIDA: IMMUNOLOGIC H STATUS OF CHICKENS AND TURKEYS GIVEN INACTIVATED VACCINE SUMMARY 12 INTRODUCTION 13 MATERIALS AND METHODS 15 RESULTS 18 DISCUSSION 20 REFERENCES, CITED 26 PART II: PASTEURELLA MULTOCIDA: ANTIBODY MEDIATED 27 RESISTANCE TO VIRULENT CHALLENGE EXPOSURE IN VACCINATED TURKEYS SUMMARY 28 INTRODUCTION 29 MATERIALS AND METHODS 30 RESULTS 34 DISCUSSION 35 ADDENDUM 38 REFERENCES CITED 42 PART III: CYCLOPHOSPHAMIDE-INDUCED IMMUNOSUPPRESSION 43 DEMONSTRATED IN PASTEURELLA MULTOCIDA VACCINATED CHICKENS SUMMARY 44 INTRODUCTION 45 iii MATERIALS AND METHODS RESULTS DISCUSSION AND CONCLUSIONS REFERENCES CITED PART IV: IN VIVO TESTS FOR DELAYED TYPE HYPERSENSITIVITY TO PASTEURELLA MULTOCIDA IN VACCINATED CHICKENS AND TURKEYS SUMMARY INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES CITED PART V: TRANSFORMATION OF CHICKEN LYMPHOCYTES OPTIMAL CULTURE CONDITIONS SUMMARY INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES CITED iv Page PART VI: STUDIES OF CELL-MEDIATED IMMUNITY 96 TO PASTEURELLA MULTOCIDA: A COMPARISON TO MY COBACTE RIUM TUB ERCULOSIS SUMMARY 97 INTRODUCTION 98 MATERIALS AND METHODS 99 RESULTS 104 DISCUSSION 106 LITERATURE CITED 124 SUMMARY AND CONCLUSIONS 126 BIBLIOGRAPHY 134 ACKNOWLEDGMENTS 140 APPENDIX 141 1 INTRODUCTION Fowl cholera is an infectious disease of poultry and wild birds which is caused by a small Gram-negative bacterium, Pasteurella multoeida. The greatest mortality is usually observed in turkeys that are older than 16 weeks of age. £. multocida can produce either chronic or acute fowl cholera. In the chronic form, P^. multocida produces localized infections in wattles, sinuses, joints, and footpads. Chronic infections are seen more often in birds that have a reduced suscepti­ bility to P^. multocida. Occasionally, chronic infections can develop into acute infections. Most acute infections have a very rapid onset; as little as twelve hours may elapse between the appearance of symptoms and death. A very rapid and overwhelming septicemia develops, with symptoms being typical of endotoxin shock. Body temperature is elevated, nasal secretions become profuse and thickened, and petechial and ecchymotic hemmorhages often occur. Torticollis, cyanosis, diarrhea and anorexia are common. An acute infection may kill thousands of birds in a single day (1). The disease is believed to be spread between birds by excretions from the mouth that contaminate feed and drinking water (2). A number of wild and domestic animals can act as carriers of avian strains of P. multocida (3). Two procedures are used currently to vaccinate against fowl cholera: a live, low virulence strain, developed at Clemson University, can be used to vaccinate turkeys through their drinking water (4,5). 2 Alternatively, turkeys or chickens are vaccinated subcutaneously with formalin-killed P^. multocida suspended in Freund's incomplete adjuvant (6,7). Neither vaccine provides complete protection against the disease. In order to produce more effective vaccines against multocida infections of poultry, the immunologic responses to vaccination need to be characterized. Humoral and cell-mediated immunologic responses have not been completely defined. Certain areas are not well-under­ stood; particularly the role of complement, the relationship of resistance to antibody titer, the effector activity of T-lymphocytes, and phagocytosis. This dissertation reports on vaccine induced immunity to P^. multocida in chickens and turkeys. The principal objective of the research was to determine: (A) the protective role of antibodies, (B) whether in vitro and in vivo correlates of cell-mediated immunity could be demonstrated, (C) whether cell-mediated imune responses to P^. multocida vaccination were of similar magnitude as responses to Mycobacterium tuberculosis vaccination. Vaccine-induced immunity was studied by various methods, including passive protection, surgical bursectomy, drug induced immunosuppression, lymphocyte transformation, migration inhibition reaction, granuloma formation, and skin testing. In this research project, a formalin-inactivated multocida bacterin was used to vaccinate chickens and turkeys. The decision to use a killed bacterin instead of a live vaccine was based on a number of 3 salient observations. (A) Lymphocytes from turkeys that received formalin-inactivated adjuvanted bacterin underwent blast transformation after exposure to 2» multocida antigens (8,9). (B) Leukocyte migration inhibition was observed when formalin- inactivated adjuvanted bacterin was used (10). (C) The Clemson University strain, a live vaccine, does not induce an immunity that is as effective or as durable as a formalin- inactivated adjuvanted bacterin (9). (D) Live vaccines occasionally induce significant mortality (5,11). (E) Live attenuated P^. miiltocida vaccines are usually given through the drinking water and are difficult to administer quantita­ tively.
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