The immunomodulatory protein RH36 is relating to blood-feeding success and oviposition in hard ticks
The immunomodulatory protein RH36 is relating to blood-feeding success and oviposition in hard ticks
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免疫调节蛋白 RH36 与硬蜱的吸血成功和产卵有关
DOI：
10.1016/j.vetpar.2017.03.017
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发表时间：
2017
影响因子：
2.6
通讯作者：
Zhou Jinlin
中科院分区：
文献类型：
--
作者：
Wang Fangfang;Lu Xiaojuan;Guo Fengxun;Gong Haiyan;Zhang Houshuang;Zhou Yongzhi;Cao Jie;Zhou Jinlin
在血蜱中鉴定出一种免疫调节蛋白RH36。RH36的cDNA序列长844 bp，编码一个分子量为24 kDa的推断蛋白。生物信息学分析表明，RH36与来自安德氏革螨的免疫调节蛋白p36相似度为34.36%。重组RH36 （rRH36）在Sf9昆虫细胞中表达可抑制脾细胞t淋巴细胞丝裂原驱动蛋白的体外增殖和IL-2、IL-12、TNF-α等多种细胞因子的表达。此外，接种rRH36的小鼠脾细胞增殖明显低于对照小鼠，表明rRH36可以直接抑制体内免疫反应。此外，对脾细胞的微阵列分析表明，rRH36下调了几个免疫调节基因的表达。RNAi沉默RH36基因后，兔耳内24 h蜱的黏附率下降37.5%，接种RH36后，蜱的黏附率下降53.06%。出乎意料的是，RNAi诱导了排卵率、充血后第12天卵巢重量和卵孵化率的显著降低。利用重组蛋白进行疫苗接种试验，进一步证实了RH36对血采和产卵的影响。这些结果表明，RH36是免疫抑制蛋白的新成员，影响蜱的吸血和产卵。
An immunomodulatory protein designated RH36 was identified in the tickRhipicephalus haemaphysaloides. The cDNA sequence of RH36 has 844 bp and encodes a deduced protein with a predicted molecular weight of 24 kDa. Bioinformatics analysis indicated that RH36 presented a degree of similarity of 34.36% with the immunomodulatory protein p36 from the tickDermacentor andersoni.The recombinant RH36 (rRH36) expressed in Sf9 insect cells suppressed the T-lymphocyte mitogen-drivenin vitroproliferation of splenocytes and the expression of several cytokines such as IL-2, IL-12, and TNF-α. Furthermore, the proliferation of splenocytes isolated from rRH36-inoculated mice was significantly lower than that in control mice, suggesting that rRH36 could directly suppress immune responsesin vivo. In addition, microarray analysis of splenocytes indicated that the expression of several immunomodulatory genes was downregulated by rRH36. The silencing of the RH36 gene by RNAi led to a 37.5% decrease in the tick attachment rate 24 h after placement into the rabbit ears, whereas vaccination with RH36 caused a 53.06% decrease in the tick engorgement rate. Unexpectedly, RNAi induced a significant decrease in the oviposition rate, ovary weight at day 12 after engorgement, and egg-hatching rate. The effects of RH36 on blood feeding and oviposition were further confirmed by vaccination tests using the recombinant protein. These results indicate that RH36 is a novel member of immunosuppressant proteins and affects tick blood feeding and oviposition.