Cloning, expression and immunoprotective efficacy of rHaa86, the homologue of the Bm86 tick vaccine antigen, from Hyalomma anatolicum anatolicum
Cloning, expression and immunoprotective efficacy of rHaa86, the homologue of the Bm86 tick vaccine antigen, from Hyalomma anatolicum anatolicum
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DOI：
10.1111/j.1365-3024.2008.01082.x
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发表时间：
2009-03-01
影响因子：
2.2
通讯作者：
Ghosh, S.
中科院分区：
文献类型：
--
作者：
Azhahianambi, P.;De la Fuente, J.;Ghosh, S.
克隆了Hyalomma anatolicum anatolicum Izatnagar分离物的Bm86同源物，并在甲基营养酵母巴斯德毕赤酵母中以细胞内、糖基化和颗粒形式表达。它被命名为rHaa86，是H. a.的第一个重组蛋白。安纳托利库姆。 Haa86 中预测的七个表皮生长因子样结构域在结构上与其 Bm86 对应物相似。 Bm86和Haa86相应的EGF样结构域之间的同一性范围为51.3%至78.3%。 rHaa86 的分子量为 120-140 kDa，可能有 50-70 kDa 糖基化。对纯化的 rHaa86 进行了免疫学表征，并评估了其在三组杂交小牛中对抗同源攻击感染的免疫保护潜力。 H. a. 女性的立即拒绝百分比饲喂免疫小牛（第 1 组）、辅助对照小牛（第 2 组）和未治疗小牛（第 3 组）时，阿纳托利菌的比例分别为 5%、12.4% 和 10.1%。喂食免疫小牛的雌性蜱的排斥率分别比喂食对照组 2 和 3 的蜱高 24.1% 和 26.4% (P < 0.05)。雌性数量、平均卵重、成年雌性和免疫原功效的减少分别为58.0%、9.0%、5.0%和61.6%。喂养第 1 组小牛的雌性的平均繁殖指数显着低于喂养对照组的雌性 (P < 0.05)，并且第 1 组小牛的饱胀幼虫数量减少了 44%。数据表明，rHaa86抗原疫苗可以作为幽门螺杆菌综合控制的有效成分之一。安纳托利库姆。
The Bm86 homologue of Hyalomma anatolicum anatolicum Izatnagar isolate was cloned and expressed in methylotropic yeast Pichia pastoris as intracellular, glycosylated and particulated form. It was named as rHaa86, the first recombinant protein of H. a. anatolicum. Seven epidermal growth factor-like domains predicted in Haa86 were structurally similar with that of its Bm86 counterpart. The identity between the corresponding EGF like domains of Bm86 and Haa86 were ranging from 51.3% to 78.3%. The molecular weight of the rHaa86 was 120-140 kDa, with possible 50-70 kDa glycosylation. The purified rHaa86 was characterized immunologically and evaluated for its immunoprotective potential against homologous challenge infestation in three groups of cross-bred calves. The immediate rejection percentage of females of H. a. anatolicum was 36 5%, 12.4% and 10.1% fed on immunized (group 1), adjuvant control (group 2) and untreated control (group 3) calves, respectively. The percent rejection of female ticks fed on immunized calves was 24.1% and 26.4% higher than for the ticks fed on control groups 2 and 3, respectively (P < 0.05). The reduction of number of females, mean weight of eggs, adult females and efficacy of immunogen were 58.0%, 9.0%, 5.0% and 61.6%, respectively. The mean reproductive index of females fed on group 1 calves was significantly lower (P < 0.05) than the females fed on the control groups and 44% reduction in the number of engorged larvae was recorded from the group 1 calves. The data demonstrated that rHaa86 antigen based vaccine could serve as one of the effective components in the integrated control of H. a. anatolicum.