用含有预测 B 细胞表位的 Rhipicephalus microplus 甲壳素酶多肽对牛进行免疫,可降低蜱的生物适应性。

IF 2.1 3区 医学 Q2 PARASITOLOGY Parasitology Pub Date : 2024-08-01 Epub Date: 2024-02-05 DOI:10.1017/S0031182024000143
María Martina Esperanza Perez-Soria, Daniel Gustavo López-Díaz, Rafael Jiménez-Ocampo, Gabriela Aguilar-Tipacamú, Massaro W Ueti, Juan Mosqueda
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引用次数: 0

摘要

牛热蜱(Rhipicephalus microplus)是影响全球畜牧业的最重要的体外寄生虫。过度依赖化学疗法控制蜱虫已导致抗杀螨剂蜱虫的出现和环境污染。以疫苗为基础的免疫策略为蜱虫控制提供了另一种方法。要开发新型蜱疫苗,关键是要确定和评估能够对牛产生保护作用的抗原。几丁质酶是一种酶,能在蜕皮时降解老的几丁质,从而实现间隙变态。在这项研究中,我们鉴定了一种 R. microplus 甲壳素酶,并评估了它降低以免疫牛为食的蜱的适应性的能力。首先,确定了 4 个分离物的预测氨基酸序列,并通过生物信息学分析了它们的相似性。设计了四种含有预测 B 细胞表位的肽。每种肽的免疫原性通过接种 2 头牛、间隔 21 天接种 4 次进行评估,并通过间接 ELISA 验证抗体反应。对具有免疫原性的多肽进行了挑战实验。甲壳素酶基因已成功扩增并测序,可与参考菌株进行比较。值得注意的是,序列之间的同一性和相似性分别达到 99.32% 和 99.84%。此外,Western 印迹检测还证明了对原生蛋白的识别。几丁质酶肽 3 能降低噬咬蜱的重量和产卵率,并能降低幼虫的活力,其效力为 71%。因此,几丁质酶肽 3 是一种可行的候选疫苗,有望将其整合到针对小尾寒羊的多抗原疫苗中。
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Immunization of cattle with a Rhipicephalus microplus chitinase peptide containing predicted B-cell epitopes reduces tick biological fitness.

Rhipicephalus microplus, the cattle fever tick, is the most important ectoparasite impacting the livestock industry worldwide. Overreliance on chemical treatments for tick control has led to the emergence of acaricide-resistant ticks and environmental contamination. An immunological strategy based on vaccines offers an alternative approach to tick control. To develop novel tick vaccines, it is crucial to identify and evaluate antigens capable of generating protection in cattle. Chitinases are enzymes that degrade older chitin at the time of moulting, therefore allowing interstadial metamorphosis. In this study, 1 R. microplus chitinase was identified and its capacity to reduce fitness in ticks fed on immunized cattle was evaluated. First, the predicted amino acid sequence was determined in 4 isolates and their similarity was analysed by bioinformatics. Four peptides containing predicted B-cell epitopes were designed. The immunogenicity of each peptide was assessed by inoculating 2 cattle, 4 times at 21 days intervals, and the antibody response was verified by indirect ELISA. A challenge experiment was conducted with those peptides that were immunogenic. The chitinase gene was successfully amplified and sequenced, enabling comparison with reference strains. Notably, a 99.32% identity and 99.84% similarity were ascertained among the sequences. Furthermore, native protein recognition was demonstrated through western blot assays. Chitinase peptide 3 reduced the weight and oviposition of engorged ticks, as well as larvae viability, exhibiting a 71% efficacy. Therefore, chitinase 3 emerges as a viable vaccine candidate, holding promise for its integration into a multiantigenic vaccine against R. microplus.

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来源期刊
Parasitology
Parasitology 医学-寄生虫学
CiteScore
4.80
自引率
4.20%
发文量
280
审稿时长
3-8 weeks
期刊介绍: Parasitology is an important specialist journal covering the latest advances in the subject. It publishes original research and review papers on all aspects of parasitology and host-parasite relationships, including the latest discoveries in parasite biochemistry, molecular biology and genetics, ecology and epidemiology in the context of the biological, medical and veterinary sciences. Included in the subscription price are two special issues which contain reviews of current hot topics, one of which is the proceedings of the annual Symposia of the British Society for Parasitology, while the second, covering areas of significant topical interest, is commissioned by the editors and the editorial board.
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