T-cell-line-tropic human immunodeficiency virus type 1 that is made resistant to stromal cell-derived factor 1alpha contains mutations in the envelope gp120 but does not show a switch in coreceptor use - PubMed (original) (raw)

T-cell-line-tropic human immunodeficiency virus type 1 that is made resistant to stromal cell-derived factor 1alpha contains mutations in the envelope gp120 but does not show a switch in coreceptor use

D Schols et al. J Virol. 1998 May.

Abstract

The NL4.3 T-cell-line-tropic human immunodeficiency virus type 1 strain is sensitive to the CXC chemokine stromal cell-derived factor 1alpha (SDF-1alpha), the natural ligand for CXC chemokine receptor 4 (CXCR4); the 50% inhibitory concentration (IC50) in MT-4 cells is 130 ng/ml. We generated resistant virus through passaging of the virus in the presence of increasing concentrations of SDF-1alpha. After 24 passages, the virus was no longer sensitive to SDF-1alpha (SDF-1alpha(res) virus) (IC50, >2 microg/ml) and became resistant to SDF-1beta (IC50, >2 microg/ml) and to a specific CXCR4 monoclonal antibody (IC50, >20 microg/ml). The SDF-1alpha(res) virus was about 10-fold less sensitive than the wild-type virus to the bicyclam AMD3100, a specific CXCR4 antagonist. The SDF-1alpha(res) virus contained the following mutations in the gp120 molecule: N106K in the V1 loop; S134N and F145L in the V2 loop; F245I in the C2 loop; K269E, Q278H, I288V, and N293D in the V3 loop; a deletion of 5 amino acids (FNSTW) at positions 364 to 368 in the V4 loop; and R378T in the CD4 binding domain. Replication of the NL4.3 wild-type virus and the SDF-1alpha(res) virus was demonstrated in U87 cells that coexpressed CD4 and CXCR4 (U87.CD4.CXCR4) but not in U87.CD4.CCR5 cells. Thus, the resistant virus was not able to switch to the CC chemokine receptor 5 (CCR5) coreceptor (the main coreceptor for macrophage-tropic viruses). The SDF-1alpha(res) virus replicated in HOS.CD4 cells expressing CCR1, CCR2b, CCR3, CCR4, CCR5, and CXCR4 but also in HOS.CD4.pBABE cells. However, all HOS transfectant cells expressed a low level of CXCR4. Neither of the two virus strains was able to infect HOS.CXCR4 or HOS.CCR5 transfectants, demonstrating the necessity of the CD4 receptor. The T-cell-line-tropic SDF-1alpha(res) virus was thus able to overcome the inhibitory effect of SDF-1alpha through mutations in gp120 but still needed CXCR4 to enter the cells.

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Figures

FIG. 1

FIG. 1

Effect of SDF-1α (1 μg/ml) on NL4.3 wild-type, SDF-1αres (passage 10), and SDF-1αres (passage 24) HIV-1 replication in MT-4 cells, as monitored by CD4 expression. Cells were infected with the virus strains at 100 CCID50 in the presence or absence of SDF-1α and stained 5 days after infection with MAb Leu-3a directly labeled with phycoerythrin (Becton Dickinson). As a control, uninfected cells also were stained with MAb Leu-3a. The percentage of CD4+ cells (uninfected cells) is indicated in each histogram.

FIG. 2

FIG. 2

Effect of SDF-1α (1 μg/ml) on NL4.3 wild-type (A, B, and C) and SDF-1αres (D, E, and F) HIV-1 replication in MT-4 cells, as monitored by anti-gp120 MAb binding. Cells were infected with the virus strains at 100 CCID50 and stained 4 days after infection with MAb NEA9305. The percentage of gp120-positive cells (HIV-1-infected cells) is indicated in each histogram. In panels A and D, cells were stained with the secondary antibody only.

FIG. 3

FIG. 3

Replication of SDF-1αres virus in U87 cells. U87.CD4.CXCR4 and U87.CD4.CCR5 cells were infected with NL4-3 wild-type (A) and SDF-1αres (B) HIV-1 in the presence of different concentrations of SDF-1α (500, 100, and 20 ng/ml). p24 antigen levels were measured 8 days after infection.

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