Analysis of long-term viral expression in CEM cells persistently infected with non syncytium-inducing HIV-1 strains.
CEM cells were infected with three HIV-1 non syncytium-inducing (NSI) strains obtained from AIDS patients or seropositive individuals. The surviving cells were followed for several months in the persistently infected cultures designated 65870/CEM, 65871/CEM and 3929/CEM, and analyzed for virus expression using light and electron microscopy, immunofluorescence, reverse transcriptase assay, polymerase chain reaction amplification (PCR), nucleic acid hybridization and flow cytometry. The virus isolates induced relatively few syncytia and other cytopathic effects in the corresponding cell lines and the number of cells positive for virus expression never rose above 44%. Distinct peaks of antigen-positive cells were obtained, coincident with high levels of reverse transcriptase activity. The cultures were strongly resistant to superinfection by laboratory strain Lai, with the exception of 65870/CEM which expressed HIV antigens in up to 15% of the cells for a few days. However, cell lysis was minimal in all cases. After long-term cultivation of the three cultures, no antigen-positive cells were detected and no trace of virus expression could be observed. The remaining cells consisted entirely of CD4-negative cells. PCR analyses indicated that cells harboring a provirus were progressively eliminated from the cultures, leaving only virus-free cells. In this system, cells carrying a latent provirus survive for a limited period of time before virus activation induces cell lysis. These results suggest that at least three types of cells exist in the CEM cell line: CD4-positive cells which are rapidly killed by the virus, a second type harboring a latent viral genome after the infection and which grow normally until activation of the resident genome by external or internal signal(s), and a third type which represents rare CD4-negative cells present in the initial CEM population and which are selected for by the NSI isolates. This is the first study documenting specific interactions between NSI strains of HIV-1 and distinct subpopulations of CEM cells grown as a single cell culture.
Yelle
J
J
Centre de recherche en virologie, Institut Armand-Frappier, Laval, Québec, Canada.
Morisset
R
R
Thibodeau
L
L
D000163:Acquired Immunodeficiency Syndrome / Q000821:virology*; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes; D002460:Cell Line; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis / Q000302:isolation & purification; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D015726:Giant Cells; D015488:HIV Antigens / Q000096:biosynthesis*; D006679:HIV Seropositivity / Q000821:virology; D015497:HIV-1 / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D007700:Kinetics; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D013997:Time Factors; D014771:Virion / Q000378:metabolism / Q000648:ultrastructure
D017931:DNA Primers; D004279:DNA, Viral; D015488:HIV Antigens