Unintegrated human immunodeficiency virus type 1 DNA in chronically infected cell lines is not correlated with surface CD4 expression

Abstract

Using a polymerase chain reaction-based assay on total cell Ijsates, we have detected unintegrated human immunodeficiency virus type 1 (HIV-1) DNA in chronically infected T-lymphocytic (ACH-2, J1) and promyelocytic (OM-10.1) cell lines. Treatment with 3'-azido-3'-deoxythymidine (AZT) or soluble CD4 inhibited accumulation of unintegrated viral DNA about 10-fold within 72 h; removal of AZT permitted recovery to pretreatment levels within 72 h. Our results indicate that unintegrated HIV-1 DNA is unstable in these cell lines and originates from a continuous process of reinfection. OM-10.1 cells had relatively high levels of surface CD4 by flow cytometry and high levels of unintegrated viral DNA by polymerase chain reaction. ACH-2 cells had very low levels of both surface CD4 and unintegrated viral DNA. However, Jl cells, with surface CD4 below the level of detection of flow cytometry had a high level of unintegrated viral DNA similar to that of OM-10.1 cells. This implies that the number of CD4 receptors is not rate limiting for reinfection.

Homologous interference resulting from the presence of defective particles of human immunodeficiency virus type 1

“…The same studies also demonstrate that defective viruses negatively affect or abolish reinfection events in a manner similar to that of AZT. This finding is in accord with a previous work which demonstrated that in OM-10.1 cells, AZT led to a reduction in integrated viral DNA probably by a mechanism of inhibition of reinfection events (6). We further investigated the basis for the inhibitory effect mediated by defective particles and determined that neither modulation of CD4 nor changes in cellular proliferation are responsible for the observed phenomenon.…”

Oscillation of the human immunodeficiency virus surface receptor is regulated by the state of viral activation in a CD4+ cell model of chronic infection

“…Because OM-10.1 cells remain CD4+ until viral activation, the possibility exists for superinfection of nonproducing cells by virions released from the small percentage of cells constitutively expressing HIV-1. An acute superinfection of OM-10.1 cells is responsible for an accumulation of extrachromosomal HIV-1 DNA (1), as demonstrated in other HIV-1-infected cell systems (1,32,33,37). Azidothymidine treatment of OM-10.1 cultures prevents the accumulation of unintegrated HIV-1 DNA (1) without affecting the activation of the integrated HIV-1 provirus by TNF-a (2,34).…”

Infection of macrophages with lymphotropic human immunodeficiency virus type 1 can be arrested after viral DNA synthesis

“…PBL were isolated and stimulated with PHA-IL2, and MDM were cultured for 3 days prior to infection with DNase treated HTLV-IIIB. MDM (1) were infected at an MOI of 3, and MDM (2) were infected at an MOI of 1. Cells were harvested at the indicated time points and subjected to PCR under standard conditions (9) using primers noted in the text.…”