本文選題：豬　切入點：弓形蟲　出處：《甘肅農(nóng)業(yè)大學(xué)》2010年碩士論文

【摘要】：弓形蟲(Toxoplasma gondii)為專性細(xì)胞內(nèi)寄生原蟲,呈全球性分布,引起的弓形蟲病是一種危害十分嚴(yán)重的人獸共患寄生蟲病,對人類健康、優(yōu)生優(yōu)育和畜牧業(yè)發(fā)展造成嚴(yán)重威脅。因此,弓形蟲病疫苗的研制已成為該病防治的當(dāng)務(wù)之急。 為闡明弓形蟲代謝分泌抗原(E/SA)對豬的免疫效應(yīng)機制,將豬隨機分為6組,每組5只,分別為E/SA組、E/SA+CpG(未乳化)組、E/SA+CpG組、E/SA +IL-2組、E/SA+IL-2+CpG組及空白對照組。共免疫2次,間隔2周,在第2次免疫后2周用弓形蟲GJS株速殖子攻擊感染。分別于免疫前、免疫后2周、免疫后4周、攻毒后1周及攻毒后2周采集仔豬外周血,用IHA檢測試劑檢測血清中抗弓形蟲抗體水平;用ELISA方法檢測血清中IL-2、IL-4、IFN-γ及TNF-α的含量;用流式細(xì)胞儀檢測外周血中CD4+及CD8+ T淋巴細(xì)胞亞群的動態(tài)變化規(guī)律。結(jié)果表明,免疫后各免疫組抗弓形蟲IHA抗體水平大部分升至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組IL-2、IL-4、IFN-γ含量顯著提高,其含量分別由免疫前的105.05、105.33、67.50 pg/mL升高到免疫后的194.03、342.50、296.50 pg/mL,與對照組差異顯著(P0.05);TNF-α含量變化不明顯。免疫組外周血CD4+、CD8+T淋巴細(xì)胞在免疫后升高,其中E/SA組升高最為明顯,CD4+、CD8+T細(xì)胞的水平分別由免疫前的21.51%,23.9%升高到免疫后的35.72%,32.8%。用弓形蟲GJS強毒攻擊后,免疫組和對照組豬體溫差異明顯,免疫組豬呈一過性低燒,食欲正常;對照組仔豬呈稽留高溫,食欲不振,并且伴有拉稀癥狀。 研究表明,用E/SA免疫豬后可刺激機體抗弓形蟲抗體產(chǎn)生,促進(jìn)IL-2、IL-4、IFN-γ的分泌,誘導(dǎo)機體CD4+、CD8+T淋巴細(xì)胞大量增殖,產(chǎn)生較強的體液免疫和細(xì)胞免疫應(yīng)答,激發(fā)宿主抗弓形蟲感染的免疫應(yīng)答反應(yīng)。
[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- 緯 and TNF- 偽 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- 緯 in serum increased to different degrees, and the level of IL-2 / IL-4 / IFN- 緯 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- 偽 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- 偽 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- 緯, 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.
【學(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R392

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