环形泰勒虫新疆喀什株抗布帕伐醌分析

Analysis of Theileria annulata Xinjiang Kashi strain resistant to buparvaquone

为了解我国是否存在环形泰勒虫耐受布帕伐醌虫株及导致对该药耐受的原因,使用布帕伐醌处理环形泰勒虫不同地方株转化的细胞TaDC(新疆喀什株)和TaNM1(内蒙古株),观察转化细胞的增殖速度、细胞内虫体情况、环形泰勒虫Ta SP基因转录及宿主细胞的细胞周期,确定药物的处理效果并分析比对布帕伐醌靶基因Ta PIN1蛋白氨基酸序列。结果显示,与TaNM1细胞相比,TaDC细胞的增殖速度受布帕伐醌影响较小;环形泰勒虫Ta SP基因的转录水平与TaDC细胞空白对照组相比略低。细胞周期检测结果显示,TaDC细胞G0/G1、G2/M和S期细胞比例相对于对照组无显著差异。氨基酸序列分析发现,新疆喀什株TaPIN1存在5个氨基酸突变位点(V24I,P26A,T53A,K121E,G124E),与国外报道的耐布帕伐醌虫株的TaPIN1蛋白具有相似的突变位点。本研究证实环形泰勒虫新疆喀什株为布帕伐醌耐药虫株,并确定该虫株Ta PIN1基因的突变是其产生耐药性的主要原因。本研究结果为我国环形泰勒虫病临床治疗和药物应用提供了数据参考。

In order to investigate whether there has buparvaquone(BW)-resistant strain of Theileria annulata in China and the caused reason,two T.annulata transformed cell lines,Ta DC(infected with T.annulata Xinjiang Kashi strain)and Ta NM1(infected with T.annulata Inner Mongolia strain)were treated with BW.And then the cell proliferation rate,the distribution of intracellular parasites,the transcription level of Ta SP gene and the cell cycle of host cells were detected at different time-point post treatment.In addition,the sequence characteristics of Ta PIN1 which is the BW target gene were analyzed.The results showed that the proliferation rate of Ta DC cells was less affected,and the transcription level of Ta SP gene was slightly lower compared to that from Ta NM1 cells.Cell cycle analysis results showed that the cells proportion of G0/G1,G2/M and S phase had no significant differences between Ta DC cells and the control group.From the amino acid sequences analyzed result,the Ta PIN1 gene of Xinjiang Kashi strain has 5 amino acid mutations(V24I,P26A,T53A,K121E,G124E)which are the major reason to BW resistance.In conclusion,we confirmed T.annulata Xinjiang Kashi strain is BW-resistant strain.The results will provide reference data for the clinical treatment and drug application for T.annulata infection in China.