Jurkat细胞
荧光素酶
病毒学
生物
交易激励
绿色荧光蛋白
转染
病毒
细胞培养
细胞生物学
分子生物学
转录因子
T细胞
遗传学
基因
免疫系统
作者
Sandrine Alais,Hélène Dutartre,Renaud Mahieux
出处
期刊:Methods in molecular biology
日期:2017-01-01
卷期号:: 47-55
被引量:7
标识
DOI:10.1007/978-1-4939-6872-5_4
摘要
Unlike HIV-1, HTLV-1 viral transmission requires cell-to-cell contacts, while cell-free virions are poorly infectious and almost absent from body fluids. Though the virus uses three nonexclusive mechanisms to infect new target cells: (1) MTOC polarization followed by formation of a virological synapse and viral transfer into a synaptic cleft, (2) genesis of a viral biofilm and its transfer of embedded viruses, or (3) HTLV-1 transmission using conduits. The Tax transactivator and the p8 viral proteins are involved in virological synapse and nanotube formation respectively. HTLV-1 transcription from the viral promoter (i.e., LTR) requires the Tax protein that is absent from the viral particle and is expressed after productive infection. The present chapter focuses on a series of protocols used to quantify HTLV-1 de novo infection of target cells. These techniques do not discriminate between the different modes of transmission, but allow an accurate measure of productive infection. We used cell lines that are stably transfected with LTR-GFP or LTR-luciferase plasmids and quantified Green Fluorescent Protein expression or luciferase activity, since both of them reflect Tax expression.
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