Niu Qingli, Bonsergent Claire, Rogniaux Hélène, Guan Guiquan, Malandrin Laurence, Moreau Emmanuelle
BIOEPAR, INRA, Oniris, La Chantrerie, 44307, Nantes, France; State Key Laboratory of Veterinary Biological Biology, LVRI, Lanzhou, China.
BIOEPAR, INRA, Oniris, La Chantrerie, 44307, Nantes, France.
Vet Parasitol. 2016 Dec 15;232:48-57. doi: 10.1016/j.vetpar.2016.11.013. Epub 2016 Nov 13.
Babesia sp. BQ1 (Lintan) is one of the parasites isolated from infected sheep in China that belongs to the B. motasi-like phylogenetic group. The rhoptry-associated-protein 1 (rap-1) locus in this group consists of a complex organization of 12 genes of three main types: 6 rap-1a variants intercalated with 5 identical copies of rap-1b and a single 3' ending rap-1c gene. In the present study, transcription analysis performed by standard RT-PCR demonstrated that the three different rap-1 gene types and the four rap-1a variants were transcribed by the parasite cultivated in vitro. Peptides, specific for each rap-1 type gene, were selected in putative linear B-epitopes and used to raise polyclonal rabbit antisera. Using these sera, the same expression pattern of RAP-1 proteins was found in parasites cultivated in vitro or collected from acute infection whereas only RAP-1a67 was detectable in merozoite extracts. However, ELISA performed with recombinant RAP-1a67, RAP-1b or RAP-1c and sera from infected sheep demonstrated that RAP-1a67 is the main RAP-1 recognized during infection, even if some infected sheep also recognized RAP-1b and/or RAP-1c.
巴贝斯虫属BQ1(临潭株)是从中国感染绵羊中分离出的寄生虫之一,属于莫氏巴贝斯虫样系统发育群。该群中的棒状体相关蛋白1(rap - 1)基因座由三种主要类型的12个基因组成复杂结构:6个rap - 1a变体,其间插入5个相同的rap - 1b拷贝和一个单一的3'末端rap - 1c基因。在本研究中,通过标准RT - PCR进行的转录分析表明,三种不同的rap - 1基因类型和四种rap - 1a变体在体外培养的寄生虫中进行转录。在假定的线性B表位中选择针对每种rap - 1类型基因的肽,用于制备兔多克隆抗血清。使用这些抗血清,在体外培养或从急性感染中收集的寄生虫中发现了相同的RAP - 1蛋白表达模式,而在裂殖子提取物中仅可检测到RAP - 1a67。然而,用重组RAP - 1a67、RAP - 1b或RAP - 1c以及感染绵羊的血清进行的ELISA表明,RAP - 1a67是感染期间主要被识别的RAP - 1,即使一些感染绵羊也能识别RAP - 1b和/或RAP - 1c。