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重组牛蜱胰蛋白酶抑制剂疫苗对牛的抗蜱侵害的保护免疫。

Protective immunity against tick infestation in cattle vaccinated with recombinant trypsin inhibitor of Rhipicephalus microplus.

机构信息

EMBRAPA Beef Cattle, Avenida Rádio Maia, 830 - Vila Popular - Caixa Postal 154, CEP 79106-550 - Campo Grande, MS, Brazil.

出版信息

Vaccine. 2012 Oct 19;30(47):6678-85. doi: 10.1016/j.vaccine.2012.08.066. Epub 2012 Sep 7.

Abstract

The cattle tick, Rhipicephalus microplus, is regarded as the most economically important ectoparasite of livestock globally. Control is achieved primarily through the use of acaricides. This approach is hampered by the development of resistance to commercial acaricides among cattle tick populations. Vaccination against R. microplus infestation is another technology that can be integrated for effective cattle tick control. Proteins belonging to the Kunitz-BPTI family are abundant in cattle tick salivary glands, midgut, and ovaries. These organs are attractive targets for the development of a novel cattle tick vaccine. Efficacy assessment against cattle tick infestation in bovines using a vaccine containing the recombinant form of a member of the Kunitz family from R. microplus produced in a yeast expression system is reported for the first time here. The yeast Pichia pastoris was bioengineered to produce the recombinant version of a trypsin inhibitor that is expressed in cattle tick larvae (rRmLTI). Immunization with rRmLTI afforded 32% efficacy against R. microplus. The estimated molecular weight of rRmLTI was 46 kDa. Structural homology to the native form of the larval trypsin inhibitor was documented by recognition of rRmLTI in Western-blots using polyclonal antibodies from mice immunized with cattle tick larval extract or rRmLTI. Bioinformatics analysis of the partial nucleotide and deduced amino acid sequences indicated that the rRmLTI closely resembles BmTI-6, which is a three-headed Kunitz protein present in cattle tick ovary and fat tissue.

摘要

牛蜱,也称璃眼蜱,被认为是全球范围内对家畜最重要的外寄生虫。其防治主要通过使用杀蜱剂来实现。然而,牛蜱种群对商业杀蜱剂的抗药性发展,对这种方法造成了阻碍。针对牛蜱感染的疫苗接种是另一种可以集成的有效牛蜱防治技术。属于 Kunitz-BPTI 家族的蛋白质在牛蜱的唾液腺、中肠和卵巢中含量丰富。这些器官是开发新型牛蜱疫苗的有吸引力的目标。本研究首次报道了使用酵母表达系统生产的来自 R. microplus 的 Kunitz 家族成员的重组形式的疫苗,对牛进行牛蜱感染的功效评估。通过生物工程改造酵母毕赤酵母,以生产在牛蜱幼虫中表达的重组版胰蛋白酶抑制剂(rRmLTI)。rRmLTI 免疫接种对 R. microplus 的功效为 32%。rRmLTI 的估计分子量为 46 kDa。通过使用从小鼠免疫牛蜱幼虫提取物或 rRmLTI 中获得的多克隆抗体在 Western blot 中识别 rRmLTI,证明了 rRmLTI 与天然幼虫胰蛋白酶抑制剂具有结构同源性。对部分核苷酸和推导的氨基酸序列的生物信息学分析表明,rRmLTI 与 BmTI-6 非常相似,BmTI-6 是存在于牛蜱卵巢和脂肪组织中的三叶 Kunitz 蛋白。

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