Macrophage-tropic HIV-1 Isolates Research Articles

Intrinsic resistance to T cell infection with HIV type 1 induced by CD28 costimulation.

When HIV-infected leukocytes are activated by the CD28 costimulatory receptor, HIV-1 is rapidly cleared from cultures, suggesting that costimulation can render T cells resistant to HIV-1 infection. In this study we tested the hypothesis that enhanced secretion of cytokines or chemokines could account for CD28-induced antiviral effects. In an acute infection system, resistance to infection with macrophage-tropic strains of HIV-1 was shown to be comprised of both soluble and cell-associated components. Induction of HIV-1 resistance was specific for CD28 costimulation, in that a variety of other accessory receptors, such as CD2, CD4, CD5, and MHC class I, failed to confer the antiviral resistance. The soluble component was secreted by both CD4 and CD8 T cells, was not unique to CD28 costimulation, and could be neutralized by removal of C-C chemokines (RANTES (regulated upon activation, normal T cell expressed and secreted) and macrophage inflammatory protein-1alpha and -1beta) from the culture supernatants of costimulated CD4 T cells. In contrast, CD28 stimulation of CD4 cells resulted in the specific induction of a pronounced intrinsic resistance to HIV-1 infection by macrophage tropic isolates of HIV-1.

The V3 domain of the HIV-1 gp120 envelope glycoprotein is critical for chemokine-mediated blockade of infection.

The ability of CD8 T cells derived from human immunodeficiency virus (HIV)-infected patients to produce soluble HIV-suppressive factor(s) (HIV-SF) has been suggested as an important mechanism of control of HIV infection in vivo. The C-C chemokines RANTES, MIP-1 alpha and MIP-1 beta were recently identified as the major components of the HIV-SF produced by both immortalized and primary patient CD8 T cells. Whereas they potently inhibit infection by primary and macrophage-tropic HIV-1 isolates, T-cell line-adapted viral strains tend to be insensitive to their suppressive effects. Consistent with this discrepancy, two distinct chemokine receptors, namely, CXCR4 (ref. 7) and CCR5 (ref. 8), were recently identified as potential co-receptors for T-cell line-adapted and macrophage-tropic HIV-1 isolates, respectively. Here, we demonstrate that the third hypervariable domain of the gp 120 envelope glycoprotein is a critical determinant of the susceptibility of HIV-1 to chemokines. Moreover, we show that RANTES, MIP-1 alpha and MIP-1 beta block the entry of HIV-1 into cells and that their antiviral activity is independent of pertussis toxin-sensitive signal transduction pathways mediated by chemokine receptors. The ability of the chemokines to block the early steps of HIV infection could be exploited to develop novel therapeutic approaches for AIDS.

The lymphocyte chemoattractant SDF-1 is a ligand for LESTR/fusin and blocks HIV-1 entry

Chemokines are chemotactic cytokines that activate and direct the migration of leukocytes. There are two subfamilies, the CXC and the CC chemokines. We recently found that the CXC-chemokine stromal cell-derived factor-1 (SDF-1) is a highly efficacious lymphocyte chemoattractant. Chemokines act on responsive leukocyte subsets through G-protein-coupled seven-transmembrane receptors, which are also used by distinct strains of HIV-1 as cofactors for viral entry. Laboratory-adapted and some T-cell-line-tropic (T-tropic) primary viruses use the orphan chemokine receptor LESTR/fusin (also known as fusin), whereas macrophage-tropic primary HIV-1 isolates use CCR-5 and CCR-3 (refs 7-11), which are receptors for known CC chemokines. Testing of potential receptors demonstrated that SDF-1 signalled through, and hence 'adopted', the orphan receptor LESTR, which we therefore designate CXC-chemokine receptor-4 (CXCR-4). SDF-1 induced an increase in intracellular free Ca2+ and chemotaxis in CXCR-4-transfected cells. Because SDF-1 is a biological ligand for the HIV-1 entry cofactor LESTR, we tested whether it inhibited HIV-1. SDF-1 inhibited infection by T-tropic HIV-1 of HeLa-CD4 cells, CXCR-4 transfectants, and peripheral blood mononuclear cells (PBMCs), but did not affect CCR-5-mediated infection by macrophage-tropic (M-tropic) and dual-tropic primary HIV-1.

Quinolinic acid production is related to macrophage tropic isolates of HIV-1.

We sought to determine whether the neurotoxin quinolinic acid (QUIN) was produced by macrophages or lymphocytes infected with isolates of HIV-1 with varying degrees of macrophage tropism derived from patients with varying stages of AIDS dementia complex (ADC). Highly macrophage tropic isolates and minimally macrophage tropic isolates were used to inoculate macrophages and QUIN production was measured. Similarly, QUIN production from macrophages was monitored using a purified cell free highly macrophage tropic isolate and laboratory isolates SF33 and SF2. Each of these experiments was also performed with lymphocytes. We found that macrophages infected with macrophage tropic isolates of HIV-1 led to QUIN production while lymphocytes did not produce QUIN. The ability of the HIV-1 infected macrophages to produce QUIN was related to the viral inoculum and the degree of macrophage tropism of the isolate. The severity of ADC in the patient from whom a particular isolate was derived was not per se a determining factor for QUIN production. Purified cell free ADC isolates also led to QUIN production by macrophages thereby suggesting that HIV-1 infection alone is capable of inducing QUIN production.

Cell Type-Specific Heterogeneity of the HIV-1 V3 Loop in Infected Individuals: Selection of Virus in Macrophages and Plasma

The primary targets for infection of HIV-1 are CD4 + T cells and monocyte/macrophage cells. We investigated the genotype heterogeneity of HIV-1 in distinct blood compartments, CD4 + T cells, monocytes, and plasma. Nucleotide sequences of proviral DNA from CD4 + T cells, monocyte cells, and cell-free virion RNA from plasma were determined in three HIV-1 infected individuals. We found that the heterogeneity in proviral DNA from CD4 + T cells was much higher than that found in monocyte cells or in virion RNA from plasma. These data showed that different swarms of HIV-1 exist in individually infected cells and plasma. Furthermore, predicted V3 amino acid sequences in the infected monocytes were very similar to the reported sequence of prototypic macrophage-tropic HIV-1 isolates. Our results show that macrophase-tropic virus is quite homogenous in monocytes/macrophages and has a common V3 structure in contrast to T-cell-tropic virus in vivo.

Mechanism of the Inhibitory Effect of Curdlan Sulfate on HIV-1 Infection in Vitro

To study the mechanism by which sulfated polysaccharides with 1,3-β-d-glucan as a main chain exert anti-HIV-1 activity, we analyzed the effects of curdlan sulfate (CRDS) on HIV-1 infection of SupT-1 cells and peripheral blood mononuclear cells. CRDS had no effect on virions, inhibited weakly HIV-1 attachment to cells, and had to be present for 24 hr to achieve protection. Lack of HIV-1 DNA corresponding to the gag region in cells incubated with the virus and CRDS and inhibition of infection after addition of 2′,3′-dideoxyinosine to cells treated with CRDS and HIV-1 for less than 24 hr suggest that CRDS delays events that precede and/or include reverse transcription. Analysis of the effect of CRDS on binding of HIV-1 neutralizing antibodies to gp120 demonstrated that both the continuous epitopes on the V3 loop and the discontinuous CD4 binding site of gp120 represent targets for CDRS. This interaction of CRDS with functional gp120 domains suggests that CRDS interferes with the membrane fusion process during HIV-1 infection. Concentrations of CRDS that were protective against infection with T cell- and macrophage-tropic HIV-1 isolates had less suppressive effects on T cell function in comparison with the related compound, dextran sulfate.