山羊弓形虫代谢分泌抗原免疫应答研究

发布时间：2018-06-29 19:40

  本文选题：山羊 + 弓形虫代谢分泌抗原　； 参考：《甘肃农业大学》2009年硕士论文

【摘要】： 弓形虫(Toxoplasma gondii)是一种专性细胞内寄生原虫,宿主广泛,可感染人及多种动物,该病呈全球性分布,对人类健康和畜牧业生产造成严重危害。因此,诊断方法和疫苗研究是弓形虫病防制的当务之急。 为了明确弓形虫代谢分泌抗原(E/SA)对山羊的免疫效果及其机制,本研究将山羊随机分为6组,每组3只,分别为E/SA组、E/SA+CpG组(未乳化)、E/SA+CpG组、E/SA+IL-2组、E/SA+IL-2+CpG组及阴性对照组。分别于免疫前、免疫后2周、免疫后4周、攻虫后1周及攻虫后2周采集山羊外周血并分离血清,分别采用IHA法检测抗体滴度的动态变化,ELISA法检测血清中IFN-γ、TNF-α、IL-2、IL-4的水平,同时用ANAE染色法检测外周血中T淋巴细胞的动态变化。结果显示,各免疫组免疫后抗体水平为1:256-1:1024,而阴性对照组为1:0-1:4,说明免疫后免疫组的IHA抗体水平开始上升;免疫前各免疫组的IFN-γ、TNF-α、IL-2、IL-4的含量分别为:14.00-18.75、30.91-42.20、10.75-18.30、20.40-24.60,而免疫后各免疫组的IFN-γ、TNF-α、IL-2、IL-4的含量分别为:175.40-215.60、89.95-253.80、230.40-301.50、39.20-54.20,阴性对照组分别为16.50-36.56、37.75-58.20、11.85-21.75、21.80-27.60,这表明E/SA免疫可引起机体内源性IFN-γ、TNF-α、IL-2、IL-4升高(P0.05);免疫后各免疫组的T淋巴细胞水平分别为:48-55、48-58、49-59、50-60、49-63、对照组为45-46,说明E/SA可刺激山羊机体T淋巴细胞水平上升(P0.05)。各组山羊在免疫4周后分别用弓形虫GJS强毒攻击,结果所有免疫组仅表现一过性低烧(40.1～41.1℃),1～4天恢复,食欲正常;而对照组山羊体温呈稽留高温(41.1～41.7℃),持续4～6天,食欲不振,并且有拉稀症状。该研究表明E/SA可刺激山羊产生较强的细胞免疫和体液免疫应答,具有较好的免疫保护性。该研究为山羊弓形虫疫苗的开发奠定了基础。
[Abstract]:Toxoplasma gondii (Toxoplasma gondii) is a specific protozoa, which can infect human beings and many animals in a wide range of hosts. It has a global distribution and is a serious hazard to human health and animal husbandry. Therefore, diagnostic methods and vaccine research are the top priority of Toxoplasma gondii prevention and control. In order to understand the immunological effect and mechanism of Toxoplasma gondii metabolic secretory antigen (E / SA) on goats, the goats were randomly divided into 6 groups, three in each group: E/ SA CpG group (without emulsification), Er / SA CpG group, Er SA CpG group, Er SA IL-2 CpG group and negative control group. The peripheral blood samples were collected before, 2 weeks after immunization, 4 weeks after immunization, 1 week after attack and 2 weeks after attack respectively. The serum levels of IFN- 纬 -TNF- 伪 TNF- 伪 and IL-2IL-4 in serum were detected by using IHA method to detect the dynamic changes of antibody titer. At the same time, the dynamic changes of T lymphocytes in peripheral blood were detected by Anae staining. The results showed that the antibody level of immunized group was 1: 256-1: 1024, while that of negative control group was 1: 0-1: 4, which indicated that the level of IHA antibody in immunized group began to rise. 鍏嶇柅鍓嶅悇鍏嶇柅缁勭殑IFN-纬,TNF-伪,IL-2,IL-4鐨勫惈閲忓垎鍒�涓�:14.00-18.75,30.91-42.20,10.75-18.30,20.40-24.60,鑰屽厤鐤�鍚庡悇鍏嶇柅缁勭殑IFN-纬,TNF-伪,IL-2,IL-4鐨勫惈閲忓垎鍒�涓�:175.40-215.60,89.95-253.80,230.40-301.50,39.20-54.20,闃存，

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