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Molecular characterization of a Trichinella spiralis serine proteinase

机译:Trichinella spiralis丝氨酸蛋白酶的分子表征

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The aim of this study was to investigate the biological characteristics and functions of a Trichinella spiralis serine proteinase (TsSerp) during larval invasion and development in the host. The full-length TsSerp cDNA sequence was cloned and expressed in Escherichia coli BL21. The results of RT-PCR, IFA and western blotting analyses showed that TsSerp was a secretory protein that was highly expressed at the T. spiralis intestinal infective larva and muscle larva stages and primarily located at the cuticle, stichosome and intrauterine embryos of the parasite. rTsSerp promoted the larval invasion of intestinal epithelial cells (IECs) and the enteric mucosa, whereas an anti-rTsSerp antibody impeded larval invasion; the promotion and obstruction roles were dose-dependently related to rTsSerp and the anti-rTsSerp antibodies, respectively. Vaccination of mice with rTsSerp elicited a remarkable humoral immune response (high levels of serum IgG, IgG1/IgG2a, IgE and IgM), and it also triggered both systemic (spleen) and local intestinal mucosal mesenteric lymph node (MLN) cellular immune responses, as demonstrated by a significant elevation in Th1 cytokines (IFN-γ) and Th2 cytokines (IL-4) after the spleen and MLN cells from vaccinated mice were stimulated with rTsSerp. Anti-TsSerp antibodies participated in the killing and destruction of newborn larvae via ADCC. The mice vaccinated with rTsSerp exhibited a 48.7% reduction in intestinal adult worms and a 52.5% reduction in muscle larvae. These results indicated that TsSerp participates in T. spiralis invasion and development in the host and might be considered a potential candidate target antigen to develop oral polyvalent preventive vaccines against Trichinella infection.
机译:本研究的目的是探讨Trichinella Spirlis丝氨酸蛋白酶(Tsserp)的生物学特性和功能在宿主幼虫侵袭和发育中。克隆全长TSSERP cDNA序列并在大肠杆菌BL21中表达。 RT-PCR,IFA和Western Blotting分析的结果表明,Tsserp是在T.螺旋肠梗理感染幼虫和肌肉幼虫阶段高度表达的分泌蛋白,主要位于寄生虫的角质石,嗜睡和宫内胚胎。 rtsserp促进肠上皮细胞(IECs)和肠粘膜的幼虫侵袭,而抗RTSSERP抗体受阻幼虫侵袭;促进和障碍的角色分别与RTSSERP和抗RTSSERP抗体依赖性相关。用RTSSERP接种小鼠引起了显着的体液免疫应答(高水平的血清IgG,IgG1 / IgG2a,IgE和IgM),它也引发了全身(脾)和局部肠粘膜肠系膜淋巴结(MLN)细胞免疫应答,如用RTSSERP刺激来自接种疫苗的小鼠的脾细胞因子(IFN-γ)和Th2细胞因子(IL-4)中的显着升高,用RTSSERP刺激。抗TSSERP抗体参与了通过ADCC杀死新生幼虫的杀死和破坏。用RTSSERP接种疫苗的小鼠在肠道成人蠕虫减少48.7%,肌肉幼虫减少52.5%。这些结果表明,TSSERP参与宿主中的T.螺旋体侵袭和发展,可能被认为是潜在的候选靶抗原,以开发针对Trichinella感染的口服多价预防疫苗。

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