SHIVKU-1感染猴感染晚期血浆病毒分离结果分析  

Macaques infected with SHIVKU-1 late-stage plasma virus isolation result analysis

作　　者：丛喆[1,2,3,4] 徐珮[1,2,3,4] 董志会 陈霆[1,2,3,4] 蒋虹[1,2,3,4] 魏强[1,2,3,4] 

机构地区：[1]北京协和医学院比较医学中心,北京100021 [2]中国医学科学院医学实验动物研究所 [3]卫生部人类疾病比较医学重点实验室 [4]国家中医药管理局人类疾病动物模型三级实验室

出　　处：《医学动物防制》2014年第12期1361-1364,1367,共5页Journal of Medical Pest Control

基　　金：国家十二五科技重大专项课题(2012ZX10004-501)

摘　　要：目的分离SHIVKU-1感染猴G1302V血浆中的病毒,以期得到一株具有较强感染力的X4嗜性病毒中国恒河猴适应株。方法用G1302V猴安乐死当天血浆与CEMx174细胞共培养,8周后收集培养上清及细胞,分别做p27蛋白水平及gag、gp120核酸序列测定。同时测定感染猴安乐死时血液样本中p27蛋白水平,并进行核酸序列分析及病毒毒力滴定。结果 G1302V猴血浆病毒分离阴性。血浆p27蛋白水平阴性,TZM-bl细胞测定病毒毒力阴性。血浆病毒及血细胞前病毒DNA核酸序列检测阳性,病毒嗜性未发生改变。结论 SHIVKU-1感染猴G1302V安乐死当天血浆中的病毒嗜性未发生变化,病毒的完整性较差及病毒的毒力较低是病毒分离阴性的主要原因。Objective To propagate a pathogenic X4- tropism SHIVKU- 1 stock via separation the virus from the plasma of macaques G1302 V which was infected with SHIVKU- 1. Methods Cocultured the plasma of G1302 V with CEMx174 cells and collected the cell pellet and supernatant 8 weeks later. The level of p27 antigen in both supernatant and plasma were tested. Viral RNA from EDTA- anticoagulated cell- free plasma and supernatant,proviral DNA from CEMx174 cell pellet and whole blood of G1302 V were extracted separately. A477 bp specific fragment of gag gene and 1600 bp fragment of gp120 gene were amplified and analyzed the genetic variation. The virus virulence in plasma was titrated with TZM- bl cells. Results It was failed to separate SHIVKU- 1 virus from G1302 V plasma. The level of p27 protein in plasma and virus virulence titration was also negative. On the contrary,the gag and gp120 gene detection were positive in both plasma and whole blood of G1302 V and the virus tropism did not change according to the sequence of V3 loop. Conclusions The virus tropism was unchanged during the infection. The failure of virus isolation from plasma was mainly owing to the decrease of complete virion and lower virulence of the virus.

关 键 词：SHIVKU-1 CXCR4 病毒分离 

分 类 号：R512.91[医药卫生—内科学]