Document 0699
 DOCN  M9590699
 TI    Efficiency of hiv-1 integration in cultured human cells
 DT    9509
 AU    Dimitrov DS; Englund G; Martin MA; National Cancer Institute, Bethesda,
       MD
 SO    NIH Conf Retroviral Integrase. 1995 Jan 19-20;:(Participants' abstracts
       and posters, abstract no. 1a). Unique Identifier : AIDSLINE
       AIDS/95920020
 AB    We have developed a PCR-based approach, which, in combination with a
       video imaging system, allowed the accurate quantitation of viral DNA
       synthesis following infection of T lymphocytes with HIV-1 U5 deletion
       mutants. The kinetics of viral DNA accumulation in infected cells was
       measured for the wild type HIV-1 NL4-3, a mutant with a 26 bp deletion
       at the 3' end of U5 (but excluding the 4bps at the very 3' end of U5)
       and a revertant in which the original 26 bp deletion had been further
       extended upstream to encompass a 45 bp deletion. The number of
       productively infected cells was calculated by using a recently developed
       theoretical model of the kinetics of virus spread in infected tissue
       cultures. The viral DNA concentration increased to a maximum of 5
       molecules per infected cell 11 hours after the start of infection and
       then decreased to 3 molecules per infected cell 13 hours later. There
       was no significant difference (less than 2-fold) between the levels of
       wild type, mutant and revertant DNA throughout the 24- hour observation
       period. However, relative to wild type the number of productively
       infected cells was 20-fold lower for the mutant and 5-fold lower for the
       revertant. These results suggest that the integration efficiency of the
       wild type HIV-1 in cultured cells is relatively high; in at least 1 out
       of 5 molecules viral DNA becomes integrated and acts as template for the
       production of progeny virions. The mutant and the revertant required 25
       and 100 molecules viral DNA, respectively, for integration leading to
       productive infection.
 DE    Cells, Cultured  DNA, Viral/*GENETICS/METABOLISM  HIV-1/*GENETICS  Human
       Kinetics  Mutation  Sequence Deletion  T-Lymphocytes/VIROLOGY  Virus
       Integration/*GENETICS  MEETING ABSTRACT

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