Analysis of clinical parameters and P-gp expression in total PBMC, CD4+ T lymphocytes, and CD14+ monocytes from HIV+ patientsa
No. of group | Patient code | Therapy | Viremia (copies/ml) | No. of CD4 cells/mm3 | Treatment protocol | P-gp expression | ||
---|---|---|---|---|---|---|---|---|
PBMC | CD4+ | CD14+ | ||||||
1 | CM0612 | Naive | 2,900 | 322 | 1.05 | 1.23 | 2.00 | |
GR1411 | Naive | 2,400 | 629 | 1.10 | 1.21 | 6.21 | ||
AS1512 | Naive | 2,300 | 433 | 0.99 | 1.06 | 1.75 | ||
VC1512 | Naive | 190,000 | 267 | 1.01 | 1.22 | 1.74 | ||
BG2211 | Naive | 25,000 | 630 | 1.03 | 1.00 | 1.65 | ||
2 | DF0612 | Responder | <80 | 657 | 2 NRTI + 1 NNRTI | 0.99 | 0.96 | 1.45 |
D10612 | Responder | <80 | 719 | 2 NRTI + PI | 1.05 | 1.13 | 1.22 | |
MC0612 | Responder | <80 | 808 | 2 NRTI + PI | 1.08 | 1.00 | 1.95 | |
ER1411 | Responder | <80 | 356 | 2 NRTI + 1 NNRTI | 0.88 | 0.88 | 3.55 | |
AG1512 | Responder | <80 | 420 | 2 NRTI + PI | 1.03 | 1.27 | 1.34 | |
GW1512 | Responder | <80 | 867 | 2 NRTI + PI | 1.00 | 0.99 | 1.15 | |
BA2211 | Responder | <80 | 805 | 2 NRTI + 1 NNRTI | 1.03 | 0.96 | 2.74 | |
OA2211 | Responder | <80 | 309 | 2 NRTI + PI | 0.97 | 0.96 | 2.74 | |
3 | AG0612 | Nonresponder | 12,000 | 363 | 2 NRTI + PI | 1.07 | 1.07 | 2.02 |
FL1512 | Nonresponder | 84,000 | 20 | 2 NRTI + PI | 0.53 | 0.57 | 1.38 | |
PE1512 | Nonresponder | 13,000 | 373 | 2 NRTI + PI | 1.01 | 0.98 | 1.55 | |
DT2211 | Nonresponder | 46,000 | 299 | 2 NRTI + PI | 0.98 | 1.17 | 3.31 | |
CF0612 | Nonresponder | 98,000 | 64 | 2 NRTI + 1 NNRTI | 1.00 | 1.11 | 1.49 |
↵ a Cytometric analysis of surface antigen expression was performed as previously described (12) by using the following monoclonal antibodies: anti-CD14-phycoerythrin (clone M5E2; Becton Dickinson, Mountain View, Calif.), anti-CD4-phycoerythrin-Cy5 (clone RPA-T4; Becton Dickinson), and anti-P-gp-fluorescein isothiocyanate (clone 12.10), which was kindly provided by Dr. M. Cianfriglia (Rome, Italy). Briefly, 5 × 105 PBMC were washed in 1% phosphato-buffered saline, 1% bovine serum albumin, and 0.1% sodium azide and were incubated for 15 min at 4°C with monoclonal antibodies. Samples were fixed in 1% paraformaldehyde, immediately acquired by a FACScalibur flow cytometer (Becton Dickinson), and analyzed with CellQuest software (Becton Dickinson). P-gp expression was analyzed in different cellular subsets: total PBMC, CD4+ T lymphocytes, and CD14+ monocytes. Data are indicated as an intensity index, which is obtained by dividing the median fluorescence intensity seen with the P-gp-specific monoclonal antibody by that seen with the immunoglobulin G2a isotype control (p170/immunoglobulin G), as previously described (10). NRTI, nucleoside reverse transcriptase inhibitors; NNRTI, nonnucleoside reverse transcriptase inhibitor.