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微小扇头蜱Bm05br抗原的分子与功能特征

Molecular and functional characterization of Bm05br antigen from Rhipicephalus microplus.

作者信息

Alzugaray María Fernanda, Parizi Luís Fernando, Seixas Adriana, Benavides Uruguaysito, da Silva Vaz Itabajara

机构信息

Departamento de Microbiología, Facultad de Veterinaria, Universidad de la Republica, Alberto Lasplaces 1550 a 1620, Montevideo, Código postal 11600, Uruguay; Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9500, Prédio 43421, Campus do Vale, Caixa Postal 15005, Porto Alegre, RS 91501-970, Brazil.

Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9500, Prédio 43421, Campus do Vale, Caixa Postal 15005, Porto Alegre, RS 91501-970, Brazil.

出版信息

Ticks Tick Borne Dis. 2017 Feb;8(2):320-329. doi: 10.1016/j.ttbdis.2016.12.008. Epub 2016 Dec 15.

Abstract

Rhipicephalus microplus is a cattle-specific tick, causing considerable losses in the livestock industry. The identification of molecules responsible for modulation of host defenses during different parasite stages can help in the development of alternative methods, such as vaccination, to control tick infestations. Hq05, a protein of unknown function identified in the tick Haemaphysalis qinghaiensis, induced a significant protective immune response when used as a vaccine in sheep. In the present study, we investigated Bm05br, the Hq05 homologous gene from R. microplus. Besides H. qinghaiensis, Bm05br homologous found in other tick species such as Rhipicephalus annulatus, Rhipicephalus sanguineus sensu lato, Haemaphysalis longicornis and Ixodes scapularis were comparatively analyzed. Bm05br expression profile in different R. microplus tissues and life-stages was determined by qRT-PCR and Western blot. Bm05br was detected in ovaries, salivary glands and the fat body of both partially and fully engorged females. The highest transcription levels were observed in partially engorged females fat body and salivary glands. Gene knockdown by RNAi reduced egg hatching rate and the weight of tick larvae obtained from treated group, when compared to controls. These results indicate that Bm05br may be involved in R. microplus reproduction. Together with its distribution and high sequence conservation across different tick species, our data suggest Bm05br as a potential antigen for development of a multispecies anti-tick vaccine.

摘要

微小牛蜱是一种专寄生于牛的蜱虫,给畜牧业造成了巨大损失。鉴定在不同寄生虫阶段调节宿主防御的分子,有助于开发诸如疫苗接种等替代方法来控制蜱虫侵袭。Hq05是在青海血蜱中鉴定出的一种功能未知的蛋白质,在绵羊中用作疫苗时可诱导显著的保护性免疫反应。在本研究中,我们调查了微小牛蜱的Hq05同源基因Bm05br。除青海血蜱外,还对在其他蜱种如环形牛蜱、长角血蜱、血红扇头蜱和肩突硬蜱中发现的Bm05br同源物进行了比较分析。通过qRT-PCR和蛋白质免疫印迹法确定了Bm05br在微小牛蜱不同组织和生活阶段的表达谱。在部分饱血和完全饱血雌蜱的卵巢、唾液腺和脂肪体中均检测到Bm05br。在部分饱血雌蜱的脂肪体和唾液腺中观察到最高转录水平。与对照组相比,RNAi介导的基因敲低降低了处理组蜱虫幼虫的孵化率和体重。这些结果表明Bm05br可能参与微小牛蜱的繁殖。结合其在不同蜱种中的分布和高序列保守性,我们的数据表明Bm05br是开发多物种抗蜱疫苗的潜在抗原。

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