Cell fusion activity of MDMs in the presence of conditioned media from cytokine-treated MDMs
| Treatment . | Antibodies4-151 . | No. of syncytia4-150 . | |
|---|---|---|---|
| IIIB . | JR-FL (% of control) . | ||
| None | None | 4 ± 1 | 228 ± 3 (100) |
| Control CM‡ | None | 3 ± 1 | 226 ± 9 (99) |
| IFN-γ CM | None | 34 ± 4 | 155 ± 12 (67) |
| IL-6 CM | None | 31 ± 3 | 162 ± 4 (71) |
| IFN-γ CM | a-MIP-1α; a-MIP-1β | 4 ± 1 | 261 ± 33 (114) |
| IL-6 CM | a-MIP-1α; a-MIP-1β | 8 ± 1 | 221 ± 14 (96) |
| MIP-1α; MIP-1β (10 ng/mL) | None | 56 ± 2 | 114 ± 18 (50) |
| MIP-1α; MIP-1β (1 ng/mL) | None | 37 ± 3 | 182 ± 11 (80) |
| MIP-1α; MIP-1β (10 ng/mL) | a-MIP-1α; a-MIP-1β | 2 ± 1 | 229 ± 20 (100) |
| MIP-1α; MIP-1β (1 ng/mL) | a-MIP-1α; a-MIP-1β | 1 ± 1 | 230 ± 6 (100) |
| MCP 3 | None | 2 ± 2 | 225 ± 21 (98) |
| Treatment . | Antibodies4-151 . | No. of syncytia4-150 . | |
|---|---|---|---|
| IIIB . | JR-FL (% of control) . | ||
| None | None | 4 ± 1 | 228 ± 3 (100) |
| Control CM‡ | None | 3 ± 1 | 226 ± 9 (99) |
| IFN-γ CM | None | 34 ± 4 | 155 ± 12 (67) |
| IL-6 CM | None | 31 ± 3 | 162 ± 4 (71) |
| IFN-γ CM | a-MIP-1α; a-MIP-1β | 4 ± 1 | 261 ± 33 (114) |
| IL-6 CM | a-MIP-1α; a-MIP-1β | 8 ± 1 | 221 ± 14 (96) |
| MIP-1α; MIP-1β (10 ng/mL) | None | 56 ± 2 | 114 ± 18 (50) |
| MIP-1α; MIP-1β (1 ng/mL) | None | 37 ± 3 | 182 ± 11 (80) |
| MIP-1α; MIP-1β (10 ng/mL) | a-MIP-1α; a-MIP-1β | 2 ± 1 | 229 ± 20 (100) |
| MIP-1α; MIP-1β (1 ng/mL) | a-MIP-1α; a-MIP-1β | 1 ± 1 | 230 ± 6 (100) |
| MCP 3 | None | 2 ± 2 | 225 ± 21 (98) |
MDMs indicate monocyte-derived macrophages; IFN-γ, interferon γ; CM, conditioned media; IL-6, interleukin 6; MIP, macrophage inflammatory protein.
Syncytia were scored after 18 hours of coculture of the macrophages and the effector cells: TF228 cells expressing IIIB envelope or 12E1 cells infected with recombinant vaccinia virus expressing R5 envelope (vCB28, JR-FL). Data represent 3 experiments.
Anti–MIP-1α and anti–MIP-1β antibodies were used at 2 μg/mL. Antibodies were incubated with the conditioned media or with MIP-1α and MIP-1β mixtures for 1 hour at room temperature.
CM were collected from the cultures of untreated MDMs (control CM) or from the cultures of cytokine-treated MDMs (IFN-γ CM and IL-6 CM) and were added to untreated autologous MDMs 1 hour prior to the addition of effector cells expressing X4 or R5 envelopes.