Analysis of clinical parameters and P-gp expression in total PBMC, CD4+ T lymphocytes, and CD14+ monocytes from HIV+ patientsa

No. of groupPatient codeTherapyViremia (copies/ml)No. of CD4 cells/mm3Treatment protocolP-gp expression
2DF0612Responder<806572 NRTI + 1 NNRTI0.990.961.45
D10612Responder<807192 NRTI + PI1.051.131.22
MC0612Responder<808082 NRTI + PI1.081.001.95
ER1411Responder<803562 NRTI + 1 NNRTI0.880.883.55
AG1512Responder<804202 NRTI + PI1.031.271.34
GW1512Responder<808672 NRTI + PI1.000.991.15
BA2211Responder<808052 NRTI + 1 NNRTI1.030.962.74
OA2211Responder<803092 NRTI + PI0.970.962.74
3AG0612Nonresponder12,0003632 NRTI + PI1.071.072.02
FL1512Nonresponder84,000202 NRTI + PI0.530.571.38
PE1512Nonresponder13,0003732 NRTI + PI1.010.981.55
DT2211Nonresponder46,0002992 NRTI + PI0.981.173.31
CF0612Nonresponder98,000642 NRTI + 1 NNRTI1.001.111.49
  • 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.