Figure 3.

Co-cultured C11 cells mature MDDCs and promote TNF-α secretion for reactivating HIV-1. (A) Phenotype characterization of MDDCs. immature MDDCs or LPS-treated MDDCs were co-cultured with or without C11 cells as above, and MDDCs were distinguished with CD11c⁺ immunostaining and the expressions of CD83, CD86 and HLA-DR were detected with flow cytometry. The percentage of positive cells was labeled. (B) Assay for TNF-α expression. MDDCs were co-cultured with or without C11 cells for 6 h, and C11 cells stimulated with or without TNF-α (20 ng/ml) for 6 h were also prepared. TNF-α expression was detected at mRNA level by qRT-PCR and normalized with β-actin. Data were presented as mean ± standard deviation (SD). (C, D) The addition of anti-TNF-α neutralizing antibodies during C11-MDDCs co-culture abolishes HIV-1 reactivation. 20 μg/ml anti-TNF-α neutralizing antibodies or IgG control were added during C11-MDDCs co-culture for 24 h (C), and viral reactivation was detected with flow cytometry (D). One representative result from at least 3 independent repeats was shown. * P < 0.05 and *** P < 0.001 were considered as significant difference in ANOVA analysis.