| RT-SHIV中国恒河猴适应株细胞水平生物学特性分析 |
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| 引用本文： | 姚南,王卫,丛喆,金光,陶真,魏强.RT-SHIV中国恒河猴适应株细胞水平生物学特性分析[J].中国比较医学杂志,2011,21(2). |
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| 作者姓名： | 姚南 王卫 丛喆 金光 陶真 魏强 |
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| 作者单位： | 中国医学科学院医学实验动物研究所,卫生部人类疾病比较医学重点实验室,国家中医药管理局人类疾病动物模型三级实验室,北京,100021 |
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| 基金项目： | "十一五"国家科技重大专项课题，协和青年基金(RT-SHIV动物模型的建立) |
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| 摘 要： | 目的 体外增值、制备动物感染来源的RT-SHIV病毒中国恒河猴适应株,比较PBMCs和CEMx174两种细胞制备出病毒的差异,同时用TZM-bl、CEMx174、PBMC三种细胞滴定测定病毒TCID50.方法 用RT-SHIV病毒静脉感染中国恒河猴,定期采血测定血浆病毒载量,当病毒载量达高峰时采血分离外周血单核淋巴细胞(PBMCs),与正常恒河猴PBMCs或CEMx174细胞共培养,定期测定培养液中的P24抗原水平,当病毒复制达高峰期时收集培养上清,分装并冻存;测定病毒RNA载量、P24抗原浓度,滴定病毒的TCID50.结果 本研究共制备了78 mL PBMCs来源的RT-SHIV病毒和85 mL CEMx174细胞来源的RT-SHIV病毒.RT基因序列和原始序列的相似度为99%,仅在第254和265位的氨基酸发现突变.RT-SHIV(PBMC)和RT-SHIV(CEMx174)病毒载量分别为1.641×108 copies/mL和8.375×108 copies/mL,P24抗原水平分别为20.745 ng/mL和4.28 ng/mL,TZM-bl、CEMx174、PBMC细胞测定病毒的TCID50分别为3.16×105 TCID50/mL和1×104 TCID50/mL,5×102 TCID50/mL和5×105 TCID50/mL,5×102 TCID50/mL和5×103 TCID50/mL.结论 PBMCs细胞来源制备的病毒较CEMx174制备的病毒具有更高的感染性.
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| 关 键 词： | 细胞培养 |
In Vitro Characterization of RT-SHIV Adapted in Chinese Rhesus Monkeys |
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| YAO Nan,WANG Wei,CONG Zhe,JIN Guang,TAO Zhen,WEI Qiang.In Vitro Characterization of RT-SHIV Adapted in Chinese Rhesus Monkeys[J].Chinese Journal of Comparative Medicine,2011,21(2). |
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| Authors: | YAO Nan WANG Wei CONG Zhe JIN Guang TAO Zhen WEI Qiang |
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| Institution: | YAO Nan,WANG Wei,CONG Zhe,JIN Guang,TAO Zhen,WEI Qiang(Key Laboratory of Human Diseases Comparative Medicine,Ministry of Health,Institute of Medical Laboratory Animal Science,Chinese Academy of Medical Sciences,Key Laboratory of Human Diseases Animal Models,State Administration of Traditional Chinese Medicine,Beijing 100021,China) |
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| Abstract: | Objective To propagate and titrate the adapted RT-SHIV from Chinese rhesus monkey using TZM-bl,CEMx174 and PBMCs cells,and to compare the possible difference of these viruses which propagated in PBMC and CEMx174,respectively.Methods Rhesus macaques free-living in China were infected with RT-SHIV intravenously.Blood samples were collected regularly and viral loads were monitored.Peripheral blood mononuclear cells(PBMCs) were examined when the viral load reached the peak,and were co-cultured with PBMCs from u... |
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| Keywords: | RT-SHIV TCID50 |
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