Figure 2
Endogenous CCL2 neutralization does not affect HIV-1 entry in MDM. (A) MDM were treated with anti-CCL2 or control Ab (2.5 μg/ml) for 20 h and then challenged with Null-VLPs, VSV-G-VLPs or Ada-VLPs (1 μg of CAp24 equivalent per 105 cells) as described in Methods. After 2 and 4 h, the percentage of GFP+ cells was assessed by flow cytometry. Data represent mean values (+SE) of the results obtained with MDM from 4 different donors. (B) MDM were treated as in A and then challenged with VSV-G-VLPs or Ada-VLPs either in the presence or in the absence of soluble LDLR (5 μg/ml) or T20 (1 μg/ml). The results obtained with 1 out of 2 different donors analyzed are shown. (C) Fluorescence microscope analysis of MDM 2 hours after the challenge with Null-VLPs, VSV-G-VLPs or Ada-VLPs. Phase-contrast micrographs of the same fields are shown on the left. Bars mark 100 μm. (D) Confocal microscope analysis of MDM 2 hours after the challenge with Null-VLPs, VSV-G-VLPs or Ada-VLPs. Images represent a Z-projection of 5 optical sections taken near the middle of the cell nucleus. VLP signal is shown in green, while nuclei are stained with DAPI (blue). Bars mark 10 μm.