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Figure 5 | Retrovirology

Figure 5

From: Endogenous CCL2 neutralization restricts HIV-1 replication in primary human macrophages by inhibiting viral DNA accumulation

Figure 5

Endogenous CCL2 neutralization induces A3A expression in MDM. (A-B) MDM were treated with anti-CCL2 or control Ab (2.5 μg/ml). After 20 h, total RNA was extracted and A3 expression was analyzed by qPCR and expressed as 2ΔΔCt values. In A, the result from 1 representative donor of 3 tested is shown. In B, data represent mean values (+SE) of the results obtained with 6 donors. **p< 0.01 (anti-CCL2 Ab vs. nil or control Ab). (C-D) MDM were treated as in A, and A3A protein expression was detected by western blot and compared to those of freshly isolated monocytes from the same donor. In C, the result from 1 representative donor of 5 analyzed is shown. In D, the ratio of A3A to actin was determined by densitometry, and graph shows the mean of the fold relative to monocytes (+SE) of the 5 donors tested. *p< 0.05 (anti-CCL2 Ab vs. nil); ***p< 0.001 (nil vs. monocyte). (E-F) MDM were treated as in A in the presence or absence of IFN-α (1:750) or IFN-β (1:250) neutralizing serum, and A3A protein expression was detected by western blot. In E, the result from 1 representative donor of 5 tested is shown. In F, the ratio of A3A to actin was determined by densitometry, and graph shows the mean of the fold relative to nil (+SE) of the 5 donors tested. (G) MDM were treated with anti-CCL2, anti-CCL3, anti-CCL4, anti-CCL5 and control Ab (2.5 μg/ml). After 20 h, cells were lysed and A3A protein expression was detected by western blot. The result from 1 representative donor of 6 tested is shown. (H) MDM were treated as in A and then infected with HIV-1BaL. After 14 days, A3A protein was detected by western blot. The result from 1 representative donor of 6 tested is shown.

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