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Figure 2 | Virology Journal

Figure 2

From: Closing two doors of viral entry: Intramolecular combination of a coreceptor- and fusion inhibitor of HIV-1

Figure 2

Mechanism of action for BFFI. A. BFFI (■) and CCR5mAb () bind to CCR5 with similar affinity as determined by FACS analysis. B. Antiviral Potency of BFFI. BFFI (□) has a higher antiviral potency than a 1:2 mixture of CCR5mAb with T-2635 (■) or either T-2635 (▲) or CCR5mAb () alone. Antiviral potency was determined in a single cycle entry assay using virus particles pseudotyped with the envelope of the CCR5-tropic virus NL-Bal. C. Reduced antiviral activity of BFFI against a partially T-2635-resistant virus. The antiviral activity of BFFI (squares) and T-2635 (circles) against wt (filled symbols) and the partially T-2635-resistant virus 098-FIres (open symbols) was determined in an antiviral assay using replication-competent virus. D. Synchronized viral infection experiment. Cells were infected at 4°C, washed and warmed to 37°C. T-2635 (), CCR5mAb (□) or the BFFI (■) were added at indicated time points. E. Antiviral activity of BFFI against X4 viruses is dependent on CCR5 expression. Antiviral activity of BFFI against virus particles pseudotyped with the envelope of the X4-tropic virus NL4-3 was determined using MAGI cells expressing CXCR4 () or CXCR4 and CCR5 (). F. Antiviral activity of BFFI against X4 virus particles is dependent on CCR5 binding. Cells were pre-incubated for 45 minutes with the CCR5 antagonist maraviroc (MVC, □), the CCR5mAb () or medium (■), washed and then infected with virus particles pseudotyped with the envelope of the X4-tropic virus NL4-3 in presence of BFFI. G. BFFI protects CCR5-expressing CD4(+) T-cells from NL4-3 virus cytopathic effect. PBMC were infected with NL4-3 virus in presence of either T-2636 or BFFI. After 5 days of incubation, depletion of total CD4(+) T-cells, CCR5+ CD4 T-cells and CCR5- CD4 T-cells was measured by flow cytometry and displayed as the ratio of CD4 to CD8 T-cells.

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