猪弓形虫代谢分泌抗原免疫应答的研究

发布时间：2018-04-01 01:15

本文选题：猪　切入点：弓形虫　出处：《甘肃农业大学》2010年硕士论文

【摘要】：弓形虫(Toxoplasma gondii)为专性细胞内寄生原虫,呈全球性分布,引起的弓形虫病是一种危害十分严重的人兽共患寄生虫病,对人类健康、优生优育和畜牧业发展造成严重威胁。因此,弓形虫病疫苗的研制已成为该病防治的当务之急。为阐明弓形虫代谢分泌抗原(E/SA)对猪的免疫效应机制,将猪随机分为6组,每组5只,分别为E/SA组、E/SA+CpG(未乳化)组、E/SA+CpG组、E/SA +IL-2组、E/SA+IL-2+CpG组及空白对照组。共免疫2次,间隔2周,在第2次免疫后2周用弓形虫GJS株速殖子攻击感染。分别于免疫前、免疫后2周、免疫后4周、攻毒后1周及攻毒后2周采集仔猪外周血,用IHA检测试剂检测血清中抗弓形虫抗体水平;用ELISA方法检测血清中IL-2、IL-4、IFN-γ及TNF-α的含量;用流式细胞仪检测外周血中CD4+及CD8+ T淋巴细胞亚群的动态变化规律。结果表明,免疫后各免疫组抗弓形虫IHA抗体水平大部分升至1:1024;血清中IL-2、IL-4、IFN-γ的含量均不同程度的升高,其中E/SA组IL-2、IL-4、IFN-γ含量显著提高,其含量分别由免疫前的105.05、105.33、67.50 pg/mL升高到免疫后的194.03、342.50、296.50 pg/mL,与对照组差异显著(P0.05);TNF-α含量变化不明显。免疫组外周血CD4+、CD8+T淋巴细胞在免疫后升高,其中E/SA组升高最为明显,CD4+、CD8+T细胞的水平分别由免疫前的21.51%,23.9%升高到免疫后的35.72%,32.8%。用弓形虫GJS强毒攻击后,免疫组和对照组猪体温差异明显,免疫组猪呈一过性低烧,食欲正常;对照组仔猪呈稽留高温,食欲不振,并且伴有拉稀症状。研究表明,用E/SA免疫猪后可刺激机体抗弓形虫抗体产生,促进IL-2、IL-4、IFN-γ的分泌,诱导机体CD4+、CD8+T淋巴细胞大量增殖,产生较强的体液免疫和细胞免疫应答,激发宿主抗弓形虫感染的免疫应答反应。
[Abstract]:Toxoplasma gondii (Toxoplasma gondii) is a parasitic protozoa in specific cells. Toxoplasma gondii is a serious zoonotic parasitic disease, which is a serious threat to human health, eugenics and animal husbandry. The development of Toxoplasma gondii vaccine has become an urgent task in the prevention and treatment of the disease. In order to elucidate the immunological mechanism of Toxoplasma gondii metabolizing secretory antigen E / SA on pigs, pigs were randomly divided into 6 groups, 5 in each group. They were divided into E/SA group, E/ SA CpG (unemulsified) group, E / SA IL-2 group, E / SA IL-2 CpG group and blank control group. The peripheral blood of piglets were collected before immunization, 2 weeks after immunization, 4 weeks after immunization, 1 week after inoculation and 2 weeks after treatment with Toxoplasma gondii GJS Toxoplasma gondii Toxoplasma gondii Toxoplasma GJS strain Toxoplasma gondii Toxoplasma gondii Toxoplasma spp. The levels of anti-Toxoplasma antibodies in serum were detected by IHA assay, the levels of IL-2IL-4IFN- 纬 and TNF- 伪 in serum were detected by ELISA method, and the dynamic changes of CD4 and CD8 T lymphocyte subsets in peripheral blood were detected by flow cytometry. After immunization, the level of anti-Toxoplasma IHA antibody increased to 1: 1024, and the level of IL-2 / IL-4 / IFN- 纬 in serum increased to different degrees, and the level of IL-2 / IL-4 / IFN- 纬 in E/SA group was significantly higher than that in control group (P < 0.05), and the level of anti-Toxoplasma IHA antibody increased to 1: 1024 after immunization. The content of TNF- 伪 increased from 105.05 105.33 pg/mL before immunization to 194.03342.50% 296.50 PG / mL after immunization, and there was no significant difference in TNF- 伪 content between the two groups. The level of CD8 T lymphocytes in peripheral blood of the immunized group was higher than that of the control group. The level of CD4 / CD8 T cells in E/SA group was significantly increased from 21.51% before immunization to 35.722 ~ 32.8% after immunization. After Toxoplasma gondii GJS was used, the difference of body temperature between immunized group and control group was obvious. The immunized pigs showed a low fever and normal appetite. The piglets in the control group had the symptoms of delayed high temperature, poor appetite and dilation. The results showed that immunizing pigs with E/SA could stimulate the production of anti-Toxoplasma antibodies, promote the secretion of IL-2hIL-4 and IFN- 纬, induce the proliferation of CD4 CD8 T lymphocytes, and produce strong humoral and cellular immune responses. To stimulate the host immune response against Toxoplasma gondii infection.
【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R392

【参考文献】