Figure 6
Inhibition by pomegranate juice (PJ) and PJ-S21, respectively, of gp120 IIIB-CD4 complex binding to cells expressing CXCR4 coreceptors. HIV-1 IIIB gp120 (5 μg) and biotinyl-CD4 (2.5 μg) were added to 100 μl phosphate buffered saline (PBS) containing 100 μg/ml bovine serum albumin (BSA) (PBS-BSA) and PJ (final 3-fold dilution) or PJ-S21 (67 mg; corresponding to 212 μg solids from PJ adsorbed onto starch). After 1 h at 20°C, the respective mixtures were added to 106 MT-2 cells. After 30 min, the cells were washed 3 times with PBS-BSA and PE-streptavidin (a fluorescent label specific for biotin; 0.1 μg) was added. After 20 min, the cells were washed and fixed by 1% formaldehyde in PBS. Flow cytometry analysis was performed in a FACSCalibur flow cytometer (Becton Dickinson Immunocytometric Systems, San Jose, CA). The median relative fluorescence values for cells exposed to gp120-CD4; gp120-CD4 + PJ; gp120-CD4 + PJ-S21; and control cells were: 13.7; 4.0; 4.3; and 2.1, respectively.