Figure 2
Transducibility of ECs by retroviruses. pBullet-EGFP was used to generate amphotropic MMLV. HSVECs (105) were cultured until confluent in 24-well plates. HSVECs were left unstimulated (A) or activated with proinflammatory cytokines (80 ng/mL TNF-α, 80 ng/mL IFN-γ, and 80 ng/mL IL-1β) for 8 h (B and C). The cells (105) were treated with retroviral supernatants only (R), retroviral supernatant plus 8 µg/mL Polybrene (R + PB), retroviral supernatants in combination with liposomes (R + L), retroviral supernatants together with liposomes and antibodies [anti-CD71, OKT9 (R + O) (A), anti-CD62, 1.2B6 (R + E) (B), anti-CD106, 1G11 (R + V) (C), or isotype control (R + C) (A-C) mAbs]. For some experiments (as indicated), the cells were preincubated with 200 µg/mL OKT9 (+ anti-CD71) (A), 1.2B6 (+ anti-CD62) (B), 1G11 (+ anti-CD106) (C), or isotype control Ig (+ cIg) (A–C). The expression of EGFP after transduction was determined by flow cytometry. The results are expressed as percentage of EGFP-positive cells (transfection efficiency) ± SD of triplicates. These are representative of three independent experiments. Results were analyzed with ANOVA. *P < 0.05.