Figure 1.

Abortive HIV‐1 RNA induces pro‐IL‐1β processing. (A) Monocyte‐derived DCs were (co)stimulated with different TLR ligands (LTA, Poly‐I:C, LPS, or R848) and/or HIV‐1 Cap‐RNA₅₈. IL‐1β secretion in cell culture supernatant was measured after 24 h by ELISA. Responses induced by the combination of HIV‐1 Cap‐RNA₅₈ and each TLR ligand were set at 1. See also Supporting information Fig. S1 for IL‐1β secretion (pg/mL). (B) Similarly, DCs were costimulated with HIV‐1 Cap‐RNA₅₈ and TLR ligands, and mRNA was extracted every 2 h to analyze IL1B transcription by quantitative real‐time PCR. mRNA expression was relative to GAPDH. Responses induced by HIV‐1 Cap‐RNA₅₈ with TLR ligand at 8 h (LTA, R848), 4 h (poly‐I:C), and 2 h (LPS) were set at 1. (C, D) DCs were stimulated with LPS, ATP, or costimulated with LPS and HIV‐1 Cap‐RNA₅₈ for 24 h. ATP was added to the LPS alone condition for the final 4 h of stimulation. IL‐1β secretion in the supernatant was measured using ELISA with concentrations ranging from 50 to 1500 pg/mL, depending on donor variability (C). To statistically compare IL‐1β responses in different donors, IL‐1β secretion was shown as fold change compared to HIV‐1 Cap‐RNA₅₈‐induced responses (D). Data are representative of collated data of six (A) and four (B) donors, or nine donors (C, D), with each symbol representing a different donor (C) (mean ± S.D.). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, Student's t‐test.