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旋毛虫抗原性重组丝氨酸蛋白酶诱导 BALB/c 小鼠保护性免疫。

An antigenic recombinant serine protease from Trichinella spiralis induces protective immunity in BALB/c mice.

机构信息

Sun Yat-sen University Zhongshan School of Medicine, Key Laboratory of Tropical Disease Control (Sun Yat-Sen University), Ministry of Education, 74 Zhongshan 2nd Road, Guangzhou, 510080, China.

出版信息

Parasitol Res. 2013 Sep;112(9):3229-38. doi: 10.1007/s00436-013-3500-6. Epub 2013 Jul 5.

Abstract

In this study, we report the cloning and characterization of a cDNA encoding a Trichinella serine protease gene (TspSP-1.3) from GenBank. The recombinant TspSP-1.3 protein (rTspSP-1.3) was expressed in an Escherichia coli expression system and purified with Ni-affinity chromatography. Real-time quantitative PCR analysis revealed that TspSP-1.3 was expressed at significantly higher levels in muscle larvae and adult worms than in newborn larvae. TspSP-1.3 was detected in excretory-secretory proteins of Trichinella spiralis with western blotting. Immunization with the rTspSP-1.3 antigen induced humoral immune responses, which manifested as elevated specific anti-rTspSP-1.3 IgG and IgE antibodies and a mixed Th1/Th2 response. To determine whether purified rTspSP-1.3 had good antigenicity and could be a vaccine candidate for the control of T. spiralis infection, we immunized BALB/c mice with rTspSP-1.3 and subsequently challenged the mice with T. spiralis larvae. The results showed that mice vaccinated with rTspSP-1.3 exhibited an average reduction in the muscle larvae burden of 39 % relative to the control group. These results suggest that TspSP-1.3 could be a novel vaccine candidate for controlling Trichinella infection.

摘要

在这项研究中,我们从 GenBank 中报道了一个编码旋毛虫丝氨酸蛋白酶基因(TspSP-1.3)的 cDNA 的克隆和特性。重组 TspSP-1.3 蛋白(rTspSP-1.3)在大肠杆菌表达系统中表达,并通过 Ni-亲和层析进行纯化。实时定量 PCR 分析显示,TspSP-1.3 在肌肉幼虫和成虫中的表达水平明显高于新生幼虫。Western blot 检测到 TspSP-1.3 存在于旋毛虫的排泄分泌蛋白中。用 rTspSP-1.3 抗原免疫诱导了体液免疫反应,表现为特异性抗 rTspSP-1.3 IgG 和 IgE 抗体的升高和混合 Th1/Th2 反应。为了确定纯化的 rTspSP-1.3 是否具有良好的抗原性,并且可以作为控制旋毛虫感染的疫苗候选物,我们用 rTspSP-1.3 免疫 BALB/c 小鼠,然后用旋毛虫幼虫对小鼠进行攻毒。结果表明,与对照组相比,rTspSP-1.3 免疫的小鼠肌肉幼虫负荷平均减少了 39%。这些结果表明,TspSP-1.3 可能是一种控制旋毛虫感染的新型疫苗候选物。

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