Activation of a beta-galactosidase recombinant provirus: application to titration of human immunodeficiency virus (HIV) and HIV-infected cells.
A quantitative bioassay for human immunodeficiency viruses has been developed on the basis of the ability of the tat gene to transactivate the expression of an integrated beta-galactosidase gene in a HeLa-CD4+ cell line. Infection by a single virion of HIV-1 or HIV-2 corresponds to a unique blue syncytium or a cell cluster detected after fixation and addition of 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (a beta-galactosidase substrate). The number of infected lymphoid cells in a culture (stimulated human peripheral blood lymphocytes and cell lines) can also be quantified by cell-to-cell transmission of HIV into the HeLa-CD4(+)-beta-galactosidase monolayer. Infections by simian immunodeficiency viruses are similarly detected. This assay has been used to determine the dose response of drugs, the half-life of HIV at 37 degrees C, and the appearance of infectious particles after virus infection.
Rocancourt
D
D
Unité de Biologie moléculaire du Développement, Unité Associée 1148 du Centre National de la Recherche Scientifique, Institut Pasteur, Paris, France.
Bonnerot
C
C
Jouin
H
H
Emerman
M
M
Nicolas
J F
JF
D015139:Blotting, Southern; D002460:Cell Line; D002999:Clone Cells; D005696:Galactosidases / Q000235:genetics*; D006678:HIV / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D015497:HIV-1 / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D008214:Lymphocytes / Q000382:microbiology; D010957:Plasmids; D011533:Proviruses / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D015183:Restriction Mapping; D015302:Simian Immunodeficiency Virus / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D015533:Transcriptional Activation; D014162:Transfection; D014775:Virus Activation; D015215:Zidovudine / Q000494:pharmacology; D001616:beta-Galactosidase / Q000235:genetics*
D012367:RNA, Viral; D015215:Zidovudine; D005696:Galactosidases; D001616:beta-Galactosidase