精液促进猴艾滋病毒体外感染

来源 :第十一届全国病毒学学术研讨会暨第六届武汉现代病毒学国际研讨会 | 被引量 : 0次 | 上传用户:datoufangyuan
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  目的:已知人精液源性病毒增强因子(Semen-derived enhancer of viral infection,SEVI),可促进HIV体外感染.由于体内研究依赖于恒河猴模型,我们研究了人和猴来源SEVI是否能促进猴艾滋病毒(Simian immunodeficiency virus,SIV)体外感染.方法:将5,10,25,50 ug/ml的人来源SEVI以及猴来源SEVI分别与三种猴艾滋病毒SIVmac239,SIVmac251,SHIVsf162p3n在37℃下预混1小时后,分别感染TZM-b1细胞和猴PBMCs,3小时后使用PBS洗去未结合的病毒并更换培养液,继续培养两天后检测细胞以及培养液上清中病毒载量;两种来源的SEVIs(50 ug/ml)分别处理TZM-b1细胞以及猴PBMCs,3小时后使用PBS洗去未结合的病毒并更换培养液,培养6,12,18小时后检测细胞内mRNA水平变化;SIVmac239与猴来源SEVI(25 ug/ml)预混1小时后,再与绿茶酚EGCG(5,10,50uM)混合处理0.5小时,使用这些预混物分别感染TZM-b1细胞或EGCG预处理TZM-b1细胞,3小时后使用PBS洗去未结合的病毒并更换培养液,培养两天后检测检测细胞以及培养液上清中病毒载量;猴来源SEVI(1 mg/ml)与EGCG(2mM)37℃混合后,按不同时间点(0,0.5,1,2,4,6,12,24,48小时)进行刚果红染色,检测EGCG对SEVI淀粉样结构的影响.
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