摘要
本文探讨了捻转血矛线虫的谷胱甘肽过氧化物酶(HC29)体外对山羊外周血单核细胞(PBMCs)功能的影响。颈静脉采取无捻转血矛线虫感染的山羊血液,分离单核细胞后加入终浓度为5μg/mL的HC29重组蛋白进行体外培养,通过免疫荧光抗体技术观察重组蛋白与单核细胞的结合情况;HC29刺激山羊外周血单核细胞,用qPCR技术测定各试验组单核细胞中IL-2、IL-4、IL-10、IL-17、IFN-γ和TGF-βmRNA的转录水平;重组蛋白刺激单核细胞,采用CCK-8法测定细胞的增值情况;将重组蛋白与巨噬细胞共培养,测定巨噬细胞吞噬和分泌NO的情况。结果表明,重组HC29能够与外周血单核细胞结合并刺激细胞因子IL-10和IFN-γ的表达水平显著提高(P<0.01),能够引起单核细胞增值(P<0.01),巨噬细胞吞噬作用增强(P<0.01)和NO分泌增加(P<0.01)。结果表明HC29是捻转血矛线虫的一个重要抗原,主要诱导Th1类免疫反应。
Effects of glutatbione peroxidase （HC29） in Haernonchus contortus on the functions of peripheral blood mononuclear cells （ PB- MCs ） from goats in vitro were investigated. Blood samples were collected from jugular vein of H. contortus infected goats. PBMCs were separated and cultured in vitro with the recombinant protein HC29 at a final concentration of 5 μg/mL. The binding of the recombinant protein to PBMCs was observed by immunofluorescence antibody technique. After stimulated with recombinant protein, cell proliferations and transcriptional levels of IL-2, IL-4, IL-10, IL-17, IFN-γ and TGF-β mRNA were tested by CCK-8 method and real-time quantitative PCR （qPCR） , respectively. The phagocytosis and NO release of macrophage were also examined. The results showed that the recombinant HC29 could bind to PBMCs and resulted in significant production of IL-10 and IFN-γ （P〈0. 01 ） and cell proliferation （P〈0. 01 ）. It also strengthened macrophage phagocytosis （ P〈0. 01 ） and boosted NO production （ P〈0. 01 ）. In conclusion, HC29 is an important antigen in H. contortus and induce mainly Thl type immune response.
出处
《畜牧与兽医》
北大核心
2014年第5期19-23,共5页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金资助项目(31172308)
江苏省优势学科项目(PAPD)
