FIG. 3.

HIV-1 infection directly primes MDM for hyperresponsiveness to TLR4 and TLR7/8-mediated activation. The effects of IFNγ stimulation versus HIV-1 infection on LPS- and CL097-induced TNF production were compared in MDM. (A) Untreated (medium alone), IFNγ-primed (10 ng/mL for 18 h), or HIV-1 infected MDM (50–5,000 TCID₅₀ for 7 days) were left untreated or activated with 10 ng/mL LPS for 6 h. ELISA was used to measure supernatant TNF. Bars represent the mean ± SEM *p < .05 using two-way ANOVA with Bonferroni correction. (B) HIV-1-infected MDM were treated with medium alone or 10 ng/mL LPS for 4 h 7 days postinfection in the presence of 3 μg/mL Brefeldin A. Intracellular p24 and TNF were measured by flow cytometry. MFI of TNF in uninfected cells and HIV-1-infected MDM gated on either p24-negative or p24-positive populations in (B) to determine the effect of HIV-1 priming on activation by (C) LPS or (D) CL097. Bars in (C, D) represent the mean ± SEM MFI of TNF; *p < .05, ***p < .001 using two-way ANOVA with Bonferroni correction. MFI, mean fluorescence intensity; TLR, toll-like receptor; SEM, standard error of the mean.