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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
11
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pubmed:dateCreated |
1999-1-4
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pubmed:abstractText |
We describe a stable and sensitive HIV evaluation system, which discriminates HIV-specific membrane fusion and early transcription events and is suitable for high-throughput inhibitor screening. A human lymphocytic line, constitutively producing infectious HIV-1, serves as Env-positive donor. A second indicator cell line carries a silent HIV-1 LTR lacZ reporter plasmid. A bicellular cocultivation setup allows titration and standardization of "fusion-induced gene stimulation (FIGS)" events. With few manipulations aspects of fusion and/or LTR induction can be distinguished and simultaneously assayed. Anti-Env-V3 antibodies prevent fusion and subsequent lacZ induction, and a Tat-specific inhibitor blocks only lacZ induction in a dose dependent manner without affecting membrane fusion. The LTR reporter is readily activated by Tat from HIV-1, HIV-2, or SIV and it responds to exogenous Tat protein. The reporter system is sensitive enough to detect single infection events on pre-seeded layers of indicator cells, which renders it potentially useful for direct virus quantification in patients' material. Moreover, our system allows to control and normalize DNA transfection efficiencies of HIV-derived plasmids. This aspect is particularly valuable for studies of RT- and protease-inhibitors and resistances, where p24 or supernatant reverse transcriptase, otherwise standard virus readouts, can be directly affected by inhibitors or mutations.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:issn |
0304-8608
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
143
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
2109-31
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pubmed:dateRevised |
2005-11-17
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pubmed:meshHeading |
pubmed-meshheading:9856097-Anti-HIV Agents,
pubmed-meshheading:9856097-Cell Line,
pubmed-meshheading:9856097-Coculture Techniques,
pubmed-meshheading:9856097-Drug Resistance, Microbial,
pubmed-meshheading:9856097-Genes, Reporter,
pubmed-meshheading:9856097-HIV Long Terminal Repeat,
pubmed-meshheading:9856097-HIV-1,
pubmed-meshheading:9856097-HIV-2,
pubmed-meshheading:9856097-HeLa Cells,
pubmed-meshheading:9856097-Humans,
pubmed-meshheading:9856097-Simian immunodeficiency virus,
pubmed-meshheading:9856097-Transfection,
pubmed-meshheading:9856097-Virus Activation
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pubmed:year |
1998
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pubmed:articleTitle |
Dissecting the mode of action of various HIV-inhibitor classes in a stable cellular system.
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pubmed:affiliation |
Novartis Pharma, Inc., Basel, Switzerland.
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pubmed:publicationType |
Journal Article
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