Figure 4
Figure 4. DCIR affects both trans- and cis-infection pathways. (A) IM-MDDCs (1 × 106 cells) were treated with oligofectamine and then either left untreated (control) or exposed to a nonspecific siRNA and a DCIR-specific siRNA. Next, cells were either left untreated or treated with EFV. IM-MDDCs (2 × 105 cells) were pulsed with NL4–3balenv (20 ng p24) for 60 minutes at 37°C. After 3 washes with PBS to eliminate unbound virus, IM-MDDCs were cocultured with autologous CD4+ T cells at a 1:3 ratio. Cell-free culture supernatants were collected at day 2 following initiation of the coculture and analyzed for the p24 content. Data shown represent the means plus or minus SD of triplicate samples from 2 different donors and are representative of 4 separate donors. Asterisks denote statistically significant data (*P < .05; **P < .01). (B) IM-MDDCs were treated with oligofectamine and then either left untreated (control) or exposed to a nonspecific siRNA and a DCIR-specific siRNA. Cells (2 × 105) were pulsed with NL4-3balenv (20 ng p24) for 60 minutes at 37°C. After 3 washes with PBS to eliminate unbound virus, IM-MDDCs were maintained in culture for 4 days. Finally, IM-MDDCs were cocultured with autologous CD4+ T cells at a 1:3 ratio. Cell-free culture supernatants were collected at different time points (ie, 2, 4, and 6 days) and analyzed for p24 contents. Data shown represent the means plus or minus SD of triplicate samples from 2 different donors and are representative of 3 separate donors. Asterisks denote statistically significant data (*P < .05). (C) IM-MDDCs were treated with oligofectamine and then either left untreated (control) or exposed to a nonspecific siRNA and a DCIR-specific siRNA. Cells (2 × 105) were pulsed with NL4–3balenv (20 ng p24) for 2 hours at 37°C. After 3 washes with PBS to eliminate unbound virus, IM-MDDCs were maintained in complete culture medium supplemented with GM-CSF and IL-4. Cell-free culture supernatants were collected at the indicated time points and analyzed for the p24 content. Data shown correspond to the means plus or minus SD of triplicate samples from 2 different donors and are representative of 4 separate donors. Asterisks denote statistically significant data (*P < .05; **P < .01).

DCIR affects both trans- and cis-infection pathways. (A) IM-MDDCs (1 × 106 cells) were treated with oligofectamine and then either left untreated (control) or exposed to a nonspecific siRNA and a DCIR-specific siRNA. Next, cells were either left untreated or treated with EFV. IM-MDDCs (2 × 105 cells) were pulsed with NL4–3balenv (20 ng p24) for 60 minutes at 37°C. After 3 washes with PBS to eliminate unbound virus, IM-MDDCs were cocultured with autologous CD4+ T cells at a 1:3 ratio. Cell-free culture supernatants were collected at day 2 following initiation of the coculture and analyzed for the p24 content. Data shown represent the means plus or minus SD of triplicate samples from 2 different donors and are representative of 4 separate donors. Asterisks denote statistically significant data (*P < .05; **P < .01). (B) IM-MDDCs were treated with oligofectamine and then either left untreated (control) or exposed to a nonspecific siRNA and a DCIR-specific siRNA. Cells (2 × 105) were pulsed with NL4-3balenv (20 ng p24) for 60 minutes at 37°C. After 3 washes with PBS to eliminate unbound virus, IM-MDDCs were maintained in culture for 4 days. Finally, IM-MDDCs were cocultured with autologous CD4+ T cells at a 1:3 ratio. Cell-free culture supernatants were collected at different time points (ie, 2, 4, and 6 days) and analyzed for p24 contents. Data shown represent the means plus or minus SD of triplicate samples from 2 different donors and are representative of 3 separate donors. Asterisks denote statistically significant data (*P < .05). (C) IM-MDDCs were treated with oligofectamine and then either left untreated (control) or exposed to a nonspecific siRNA and a DCIR-specific siRNA. Cells (2 × 105) were pulsed with NL4–3balenv (20 ng p24) for 2 hours at 37°C. After 3 washes with PBS to eliminate unbound virus, IM-MDDCs were maintained in complete culture medium supplemented with GM-CSF and IL-4. Cell-free culture supernatants were collected at the indicated time points and analyzed for the p24 content. Data shown correspond to the means plus or minus SD of triplicate samples from 2 different donors and are representative of 4 separate donors. Asterisks denote statistically significant data (*P < .05; **P < .01).

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