三磷酸甘油醛异构酶同源蛋白的鉴定与评估：长角血蜱作为一种抗蜱感染候选疫苗。

Characterization and evaluation of a new triosephosphate isomerase homologue from Haemaphysalis longicornis as a candidate vaccine against tick infection.

机构信息

Ministry of Education Key Laboratory of Molecular and Cellular Biology, Hebei Province Key Laboratory of Animal Physiology, Biochemistry and Molecular Biology, College of Life Sciences, Hebei Normal University, No. 20 East Road of 2nd South Ring, Shijiazhuang 050024, China.

出版信息

Ticks Tick Borne Dis. 2022 Jul;13(4):101968. doi: 10.1016/j.ttbdis.2022.101968. Epub 2022 May 16.

DOI:10.1016/j.ttbdis.2022.101968

PMID:35609507

Abstract

Haemaphysalis longicornis is an obligate hematophagous ectoparasite that transmits a variety of pathogens causing life‑threatening diseases in humans and animals. Triosephosphate isomerase (TIM) is a key enzyme in glycometabolism, making in an interesting anti-tick vaccine candidate antigen. In this study, the open reading frame (ORF) of the TIM homologue from H. longicornis (HlTIM) was shown to consist of 747 bp encoding a protein of 248 amino acids. HlTIM gene expression was detected in all developmental stages and in all tissues of the unfed female tick by quantitative real-time PCR. The HlTIM gene was cloned and inserted into pET-32a (+) to obtain the recombinant HlTIM protein (rHlTIM) and its immunogenicity was confirmed by Western blot. ELISA results showed that rabbits immunized with rHlTIM produced a humoral immune response. A vaccine trial in rabbits against H. longicornis infestation demonstrated that the engorgement weight, oviposition and hatchability of ticks from the rH1TIM group was decreased by 8.6%, 35.4% and 17.3% respectively, compared to the histidine-tagged thioredoxin (Trx) control group. Considering the cumulative effect of vaccination on the evaluated parameters, results showed 50.9% efficacy in the rHlTIM group. The data reported here demonstrate that rHlTIM has potential for further development of a new candidate vaccine for tick control.

摘要

长角血蜱是一种专性吸血的外寄生虫，可传播多种病原体，导致人类和动物的生命受到威胁。磷酸丙糖异构酶（TIM）是糖代谢中的关键酶，使其成为一种有趣的抗蜱疫苗候选抗原。在本研究中，显示长角血蜱 TIM 同源物（HlTIM）的开放阅读框（ORF）由 747bp 组成，编码 248 个氨基酸的蛋白质。通过实时定量 PCR 检测到未喂食的雌性蜱的所有发育阶段和所有组织中都表达了 HlTIM 基因。克隆 HlTIM 基因并插入 pET-32a（+）中，获得重组 HlTIM 蛋白（rHlTIM），并通过 Western blot 证实其免疫原性。ELISA 结果表明，用 rHlTIM 免疫的兔子产生了体液免疫反应。在兔子中针对长角血蜱侵染的疫苗试验表明，与组氨酸标记的硫氧还蛋白（Trx）对照组相比，rH1TIM 组的蜱的饱血重量、产卵和孵化率分别降低了 8.6%、35.4%和 17.3%。考虑到接种对评估参数的累积效应，rHlTIM 组的效力为 50.9%。这里报告的数据表明，rHlTIM 具有进一步开发新型蜱控制候选疫苗的潜力。