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1、ENHANCEMENT OF IMMUNE RESPONSES TO VACCINATION AGAINST FOOT-AND-MOUTH DISEASE BY ORAL ADMINISTRATION OF EXTRACT MADE FROM COCHINCHINA MOMORDICA SEEDS (木鳖子皂甙木鳖子皂甙木鳖子皂甙木鳖子皂甙对口蹄疫疫苗的免疫增强作用研究对口蹄疫疫苗的免疫增强作用研究对口蹄疫疫苗的免疫增强作用研究对口蹄疫疫苗的免疫增强作用研究) Academic Dissertation Presented in Partial Fulfillment of the Requi
2、rements for the Award of the Degree Doctor of Philosophy (Ph.D.) In Preventive Veterinary Medicine (Microbiology and Immunology) By Kedsirin Sakwiwatkul Supervised by Professor Songhua HU, Ph.D. Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310029, Chin
3、a June 2009 UNIVERSITY CODE: 10335 STUDENT ID No.: 10617037 ENHANCEMENT OF IMMUNE RESPONSES TO VACCINATION AGAINST FOOT-AND-MOUTH DISEASE BY ORAL ADMINISTRATION OF EXTRACT MADE FROM COCHINCHINA MOMORDICA SEEDS （木鳖子皂甙木鳖子皂甙木鳖子皂甙木鳖子皂甙 对口蹄疫疫苗的免疫增强作用研究对口蹄疫疫苗的免疫增强作用研究对口蹄疫疫苗的免疫增强作用研究对口蹄疫疫苗的免疫增强作用研究) ACADEM
4、IC DISSERTATION Presented in Partial Fulfillment of the Requirements for the Award of the Degree Doctor of Philosophy (Ph.D.) CANDIDATE: KEDSIRIN SAKWIWATKUL MAJOR: PREVENTIVE VETERINARY MEDICINE SPECIALITY: MICROBIOLOGY AND IMMUNOLOGY SUPERVISOR: PROFESSOR SONGHUA HU, Ph.D. Department of Veterinary
5、 Medicine, College of Animal Sciences Zhejiang University, Hangzhou, China June 2009 THESIS RELEASE FORM The author has agreed that the library, Zhejiang University Hangzhou, China, can make this thesis freely available for inspection. Moreover, the author has agreed that permission for extensive co
6、pying of this thesis for scholarly purposes may be granted by the professor who has supervised the thesis work herein or, in their absence, by the Head of department, or the Dean of Faculty in which this work was done. It is understood that the recognition will be given to the author of this thesis
7、and the Zhejiang University Hangzhou in any use of the material in this thesis. Copying or publication or any other use of the thesis for financial gain without the prior approval by the Zhejiang University Hangzhou and the authors written permission is prohibited. Request for permission to copy or
8、to make any other use of material in this thesis in whole or in part should be addressed to: The Head of the Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University Hangzhou 310029, Peoples Republic of China . RESEARCH CERTIFICATE This is to certify that the investigation
9、entitled “ENHANCEMENT OF IMMUNE RESPONSES TO VACCINATION AGAINST FOOT-AND-MOUTH DISEASE BY ORAL ADMINISTRATION OF EXTRACT MADE FROM COCHINCHINA MOMORDICA SEEDS” was carried out by Miss Kedsirin Sakwiwatkul, candidate for the degree, Doctor of Philosophy (Ph.D.) in Preventive Veterinary Medicine (Mic
10、robiology and Immunology), under my supervision and guidance; and that the investigation is original. Supervisor. (Professor Songhua HU) The Head of the Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University Hangzhou 310029, Peoples Republic of China Financial support for
11、 present study was funded by the Department of Science and Technology of Zhejiang Province, Peoples Republic of China Project No. 2004C32047. The Research Scholar was supported by Mahasarakham University, Talad, Mahasarakham, Thailand. ABBREVIATIONS ABBREVIATIONS APC antigen presenting cells CBS car
12、bonate buffer solution CK cytokines CMI cell mediated immunity Con-A concavalin A CTL cytotoxic T lymphocytes DD double distilled water DMSO dimethyl sulfoxide ECMS extract of Cochinchina momordica seed ELISA enzyme-linked immunosorbent assay EU endotoxin unit FCS fetal calf serum FMD foot-and-mouth
13、 disease FMDV foot-and-mouth disease virus HBSS Hanks balanced salt solution HRP horse radish peroxidase IFN interferon Ig immunoglobulin IL interleukin ISCOMs immunostimulating complex LAL Limulus amebocyte lysate LPS lipopolysaccharide MTT 3-(4, 5-dimethylthiazoyl-2-yl)-2, 5-diphenyltetrazolium br
14、omide NK natural killer cell OD optical density PBS phosphate buffered saline PBST PBS with taween-20 ABBREVIATIONS QA extract of the bark of Quillaja saponaria rpm rotation per minute SC subcutaneous TNF tumor necrosis factor Th T helper cells Th1 T helper cells type 1 Th2 T helper cells type 2 TMB
15、 3, 3, 5, 5-tetramethylbenzidine Tables of Contents I Table of Contents ABBREVIATIONSI TABLE OF CONTENTS I LIST OF FIGURES.V LIST OF TABLESVII ABSTRACT.1 CHAPTER 111 GENERAL INTRODUCTION11 1.1 IMPORTANCE.11 1.2 HYPOTHESIS13 1.3 STRUCTURE OF THE STUDY14 1.4 COCHINCHINA MOMORDICA14 1.5 FRUIT15 1.6 ROO
16、T.16 1.7 SEED.16 1.8 EXTRACTION OF COCHINCHINA MOMORDICA SEEDS17 1.9 QUILLAJA SAPONINS20 1.10 STUDY ON MICE.22 1.11 THESIS OBJECTIVES.22 REFERENCES23 CHAPTER 231 NEED FOR IMPROVEMENT OF THE IMMUNE RESPONSE TO VACCINATION .31 AGAINST FMD31 2.1 INTRODUCTION31 2.2 ECONOMIC CONSEQUENCES OF FOOT-AND-MOUTH DISEASE.33 2.2.1 Local level.33 2.2.2 National level34 2.3 IMPROVING IMMUNE RESPONSE TO VACCINATION AGAINST FMD.35 REFERENCES37 CHAPTER 339
