Fig. 5: In vitro IL1B activated NF-κB, increased HIV proviral transcription, and inhibited spreading infection.

a Effects of IL1B on NF-κB activity were assessed using A549 NF-κB reporter cells. Cultures were treated with IL1B, TNFα, or LPS and infected with VSV-pseudotyped NL4-3, CH058, or Mock control. After 24 hrs the Alkaline Phosphatase Blue Microwell assay was performed with OD650 values relative to the corresponding no treatment control (NT) reflecting NF-κB expression which is shown on the y-axis. The data presents an average of n = 3 independent experiments ±SEM. Blue: IL1B, teal: TNFα, yellow: LPS. b PBMC from n = 3 participants were treated with IL1B or TNFα and examined for IκBα phosphorylation as described in the methods section. Graphs present the protein expression from these n = 3 participants as mean ± SEM. Corresponding representative uncropped images are provided in the Source Data file. White: no treatment, light blue: IL1B 10 ng/ml, medium blue: IL1B 50 ng/ml, dark blue: TNFα. c Effects of IL1B in vitro when HIV was quantified after a single round of infection. Plots show the relative proportions of pMorpheus-V5 latently (blue) or productively (orange) infected PBMC in cultures treated with IL1B prior to, simultaneously, or after transduction with Env viral particles carrying the indicated Env protein. The background obtained from uninfected cells (Mock) was subtracted from the values obtained for cells infected with the pMorpheus HIV-1 construct pseudo-typed with the indicated envelope proteins. The data represent n = 6 (X4) or n = 3 (R5 and R5X4) individual healthy participants, with error bars representing the average ±SEM. Corresponding gating strategies and representative plots are shown in Supplementary Fig. 7. d Effects of IL1B on spreading HIV-1 infection in cell culture. Using HIV-1 YU-2, bar plots display the relative p24-positive cell fractions after pre-treatment with increasing concentrations of IL1B (from 0.01-10.0 ng/ml, 10-fold increments) across n = 4 different participants. Significance was calculated using the Mann-Whitney U two-sided test. Orange: no treatment, yellow: different concentrations of IL1B treatment. e, f Bar plots display the average infectious virus yields (e) and p24 antigen levels (f) of n = 5 different participants at 4 days post-infection relative to the no IL1B treatment controls normalized to 100% ±SEM. Corresponding replication curves are shown in Supplementary Fig. 9. For all bar graphs shown, bar height and error bars represent mean values +/- SEM. Significant differences in all panels except (d), were determined using two-sided unpaired t-test analyses. Asterisks indicate statistical significance (*P < 0.05, **P < 0.01, *** P < 0.001, ****P < 0.0001). Source data and exact P values are provided in the Source Data file.