Identification of Haemaphysalis longicornis Genes Differentially Expressed in Response to Babesia microti Infection
Identification of Haemaphysalis longicornis Genes Differentially Expressed in Response to Babesia microti Infection
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DOI：
10.3390/pathogens9050378
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发表时间：
2020-05
期刊：
影响因子：
3.7
通讯作者：
Weiqing Zheng;R. Umemiya-Shirafuji;Shengen Chen;Kiyoshi Okado;P. F. Adjou Moumouni;Hiroshi Suzuki;
中科院分区：
文献类型：
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作者：
Weiqing Zheng;R. Umemiya-Shirafuji;Shengen Chen;Kiyoshi Okado;P. F. Adjou Moumouni;Hiroshi Suzuki;
长角血蜱是一种蜱虫和多种病原体的媒介，包括人类致病性微小巴贝虫。本研究的目的是利用抑制减法杂交（SSH）方法鉴定雌性长角夜蛾对微孢子虫Gray菌株感染的差异表达基因。随机选取302个与巴贝虫感染相关的正向减法SSH cDNA文库和110个反向cDNA文库进行测序分析。对蜱虫正向cDNA文库中蜱虫序列进行基因本体定位和序列分析，发现有14个基因与刺激反应或/和免疫系统过程相关，其中7个基因每千碱基（SPK）标准化序列数较高。实时荧光定量PCR检测结果显示，饲养蜱中obg样atp酶1 （ola1）、钙网蛋白（crt）、卵黄原蛋白1 （Vg1）和Vg2等8个基因表达上调。与未感染的蜱相比，感染蜱有6个基因上调，包括ola1、crt和Vg2。通过RNA干扰对饲养或感染巴贝虫的蜱中上调基因的功能分析表明，感染蜱中下调crt和Vg2，而未感染蜱中下调ola1会加速膨胀。相比之下，被感染蜱体内Vg1的敲除延迟了充血。被感染蜱体内的crt和Vg1基因被敲除后，雌虫体重增加。与对照组相比，Vg2敲除使微螺旋体感染水平降低了51%。本文报道的结果增加了我们对长角锥虫基因在血液摄食和微孢子虫感染中的作用的理解。
Haemaphysalis longicornis is a tick and a vector of various pathogens, including the human pathogenetic Babesia microti. The objective of this study was to identify female H. longicornis genes differentially expressed in response to infection with B. microti Gray strain by using a suppression subtractive hybridization (SSH) procedure. A total of 302 randomly selected clones were sequenced and analyzed in the forward subtracted SSH cDNA library related to Babesia infection, and 110 clones in the reverse cDNA library. Gene ontology assignments and sequence analyses of tick sequences in the forward cDNA library showed that 14 genes were related to response to stimulus or/and immune system process, and 7 genes had the higher number of standardized sequences per kilobase (SPK). Subsequent real-time PCR detection showed that eight genes including those encoding for Obg-like ATPase 1 (ola1), Calreticulin (crt), vitellogenin 1 (Vg1) and Vg2 were up-regulated in fed ticks. Compared to uninfected ticks, infected ticks had six up-regulated genes, including ola1, crt and Vg2. Functional analysis of up-regulated genes in fed or Babesia-infected ticks by RNA interference showed that knockdown of crt and Vg2 in infected ticks and knockdown of ola1 in uninfected ticks accelerated engorgement. In contrast, Vg1 knockdown in infected ticks had delayed engorgement. Knockdown of crt and Vg1 in infected ticks decreased engorged female weight. Vg2 knockdown reduced B. microti infection levels by 51% when compared with controls. The results reported here increase our understanding of roles of H. longicornis genes in blood feeding and B. microti infection.