Two serine protease inhibitors (serpins) that induce a bovine protective immune response against Rhipicephalus appendiculatus ticks
Two serine protease inhibitors (serpins) that induce a bovine protective immune response against Rhipicephalus appendiculatus ticks
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DOI：
10.1016/j.vaccine.2005.10.055
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发表时间：
2006-03-20
期刊：
影响因子：
5.5
通讯作者：
Onuma, M
中科院分区：
文献类型：
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作者：
Imamura, S;Namangala, B;Onuma, M
我们之前已经对两种作为抗蜱候选疫苗的扇头扇贝丝氨酸蛋白酶抑制剂（RAS-1 和 -2）进行了初步表征。在本研究中，为了阐明这一假设，我们生成并进一步表征了重组 RAS-1 和 -2（rRAS-1 和 -2），并测试了它们作为牛抗蜱鸡尾酒疫苗的效力。 RT-PCR 分析表明，RAS-1 和 -2mRNA 转录本在蜱的所有生命周期阶段都有表达，与性别无关。根据SDS-PAGE判断，考虑到表达载体产生了与18-22kDa TRX蛋白融合的重组蛋白，rRAS-1和-2分别迁移为分子量约为64和60kDa的蛋白。根据抗蜱唾液血清与 rRAS-1 和 -2 的反应性确定，RAS-1 和 -2 并未分泌到咬伤部位，这表明这两种蛋白都是隐藏抗原。给牛接种 rRAS-1 和 -2 组合疫苗可赋予牛显着的针对蜱的保护性免疫力，使若虫肿胀率降低 61.4%，成年雌性和雄性蜱的死亡率分别增加 28% 和 43%。这是第一份关于使用源自 R.appiculatus 的两种不同丝氨酸蛋白酶抑制剂组合并使用牛作为自然宿主的抗蜱疫苗试验的报告。 (c) 2005 Elsevier Ltd. 保留所有权利。
We have previously undertaken preliminary characterization of two Rhipicephalus appendiculutus serine protease inhibitors (RAS-1 and -2) as anti-tick vaccine candidates. In this study, to clarify this hypothesis, we generated and further characterized recombinant RAS-1 and -2 (rRAS-1 and -2) and tested their potency as a cocktail anti-tick vaccine in cattle. RT-PCR analysis showed that RAS-1 and -2mRNA transcripts are expressed during all life cycle stages of ticks, independent of sex. As judged by SDS-PAGE rRAS-1 and -2 migrated as a molecular weight of around 64 and 60 kDa protein, respectively, considering that the expression vector produced a recombinant protein fused with 18-22 kDa TRX protein. RAS-1 and -2 were found not to be secreted into the bite site as determined by the reactivity of anti-tick saliva sera to rRAS-1 and -2, suggesting that both proteins are concealed antigens. Vaccination of cattle with a combination of rRAS-1 and -2 conferred significant protective immunity against ticks, resulting in 61.4% reduction in nymph engorgement rate, and in 28 and 43% increased mortality rate in adult female and male ticks, respectively. This is the first report on an anti-tick vaccine trial using a combination of two different serpins derived from R. appendiculatus, and using cattle as a natural host. (c) 2005 Elsevier Ltd. All rights reserved.