Reversion of a polymerase-defective integrated HIV-1 genome.
The 8E5 clonal cell line, derived from HIV-1-infected CEM cells, carries a single, reverse transcriptase (RT)-defective copy of an integrated HIV genome. The absence of RT production is a consequence of a frame shift in the pol gene, due to the addition of a single base at position 3241. We report here that 8E5 cells produce an infectious virus that can be serially passaged on CD4+ lymphoid cells. This virus (8E5R) is RT positive, but displays a slow replication profile, together with a reduced cytopathic effect. The nucleotide sequence of a segment of the pol region produced by PCR amplification of DNA from 8E5R-infected cells shows that the single nucleotide insertion characteristic of the 8E5 genome had been corrected. The same reversion event was also found to occur in most single-cell clones derived from the 8E5 cell line. Because this cell line is used in many laboratories, notably as a standard for PCR quantitation, and is generally considered as unable to produce infectious virus, our findings should prompt investigators to use particular care in the handling of these cells.
Quillent
C
C
Département SIDA et Rétrovirus, Institut Pasteur, Paris, France.
Dumey
N
N
Dauguet
C
C
Clavel
F
F
D001483:Base Sequence; D002460:Cell Line; D003673:Defective Viruses / Q000235:genetics*; D016679:Genome, Viral; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development / Q000648:ultrastructure; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D012194:RNA-Directed DNA Polymerase / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D014764:Viral Proteins / Q000032:analysis; D016662:Virus Integration; D014779:Virus Replication
D014764:Viral Proteins; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase