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

Abstract

We have developed a unique physiologic model of chronic human immunodeficiency virus type 1 (HIV-1) infection, OM-10.1, clonally derived from infected HL-60 promyelocytes and harboring a single integrated provirus. Unlike other models of chronic infection, OM-10.1 cultures remain CD4+ under normal culture conditions, during which <10% of the cells constitutively express HIV-1 proteins. However, when treated with tumor necrosis factor alpha (TNF-a), OM-10.1 cultures dramatically increased (>35-fold) HIV-1 expression and rapidly down-modulated surface CD4, as >95% of the cells became HIV-1+. The complete loss of surface CD4 following viral activation was neither associated with apparent cytopathicity nor due to a decline of available CD4 mRNA. There was, however, a temporal association between CD4 down-modulation and the accumulation of intracellular HIV-1 gpl60/120; in addition, intracellular CD4-gpl60 complexes were identifiable in OM-10.1 cell lysates at time points following TNF-a induction after surface CD4 was no longer detectable. Surface CD4 expression by OM-10.1 cells returned once viral activation ceased and could be repeatedly oscillated upon HIV-1 reactivation. Furthermore, inhibition of protein kinase activity following maximal TNF-a stimulation of OM-10.1 cells quickly returned activated HIV-1 to a state of latency, as evidenced by an accelerated return of surface CD4. These results with the new OM-10.1 cell line demonstrate that CD4 surface expression can be maintained during chronic infection and is critically dependent on the state of viral activation, that CD4-gpl60 intracellular complexing is involved in CD4 down-modulation, and that protein kinase pathways not only function in the primary induction of latent HIV-1 but also are required for maintaining the state of viral activation.

In vitro modeling of the HIV-macrophage reservoir

“…The Spearman r, a measure of the degree of correlation between two values was 0.99 for day 7 and 0.98 for day 60 -78. The GFPϩ population (after background subtraction) was divided based on MFI into lo (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20), med (21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40) and hi (41ϩ). These three groups were then subdivided based on the MFI of p24 expression into lo (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)…”

Inhibition of interleukin‐6‐induced human immunodeficiency virus type 1 expression by anti‐GP130 monoclonal antibody

“…Cells. OM-10.1 and U1 cells (14,15), which are myelomonocytic and promonocytic cell lines latently infected with HIV-1, respectively, were maintained in RPMI-1640 medium supplemented with 10% heat-inactivated fetal calf serum, 100 U/ml penicillin G; and 100 lag/ml streptomycin (culture medium). These cell lines produce little or no HIV-1 under basal conditions but do produce significant level of virus after stimulation with various substances, including TNF-ot.…”

Detection of Acute HIV-1 Infection by RT-LAMP

“…An HIV-1 DNA linearity panel was created using the human monocytic cell line OM-10.1 [23], which contains a single integrated HIV-1 provirus per cell. DNA was extracted using a QIAamp DNA Blood Mini kit (Qiagen, Valencia, CA), as previously described [17].…”