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High frequency of isolation of defective human immunodeficiency virus type 1 and heterogeneity of viral gene expression in clones of infected U-937 cells.
- Source :
-
Journal of virology [J Virol] 1990 Apr; Vol. 64 (4), pp. 1745-55. - Publication Year :
- 1990
-
Abstract
- Limiting-dilution techniques were employed to derive single-cell clones from U-937 cells that had been chronically infected with human immunodeficiency virus type 1. All clones thus obtained were positive for the presence of viral antigens; however, not all of the clones produced infectious progeny virus, as detected by the presence of reverse transcriptase (RT) activity in culture fluids. Six of these clones were monitored over time to determine whether their phenotype of human immunodeficiency virus type 1 expression was stable. Three clones maintained production of RT activity at a high level and showed a very high percentage of cells positive for viral p24 antigen, as determined by indirect immunofluorescence. The other three clones showed variations in either their levels of RT activity or the number of cells positive for p24, after which they stabilized. Infectious virus could be recovered from only three clones, as assessed by coculture experiments with different cell types. Two other clones were shown to produce noninfectious viruses. Molecular analyses at the DNA, RNA, and protein levels showed extensive variations between the viral isolates recovered from each clone.
- Subjects :
- Blotting, Northern
Blotting, Southern
Cells, Cultured
Clone Cells microbiology
DNA, Viral metabolism
Giant Cells microbiology
HIV Antigens biosynthesis
HIV Antigens genetics
HIV-1 isolation & purification
HIV-1 pathogenicity
Humans
RNA, Messenger metabolism
RNA, Viral metabolism
RNA-Directed DNA Polymerase genetics
Viral Proteins biosynthesis
Viral Proteins genetics
Gene Expression
HIV-1 genetics
Subjects
Details
- Language :
- English
- ISSN :
- 0022-538X
- Volume :
- 64
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Journal of virology
- Publication Type :
- Academic Journal
- Accession number :
- 1690823
- Full Text :
- https://doi.org/10.1128/JVI.64.4.1745-1755.1990