Fig 1: HC69.5. HIV (GFP+) were treated with/without 100 μg/ml poly IC, 1 μM of Δ(9)-THC or CBD. Supernatants were collected after 24 h of treatment and analyzed by ELISA MAX™ Deluxe Set Human IL-1ß (Biolegend; cat# 437004). GraphPad Prism 5 software was used to graph and conduct statistical analysis. Normality distribution was checked by the Kolmogorov–Smirnov test. Then, statistical significances (p ≤ 0.05) were calculated by ANOVA followed by Dunn’s test as post-hoc test.
Fig 2: IL-1 and IFN signaling are necessary and sufficient to potentiate the response of WA1mRNA.a, Schematic showing C57BL/6J wild-type mice immunized i.m. with saline, WA1mRNA or WA1mRNA+MA on D0 (prime) and on D14 (boost) and treated with anakinra (Ana) (i.p.) or VX765 (V765) (i.p., 25 mg kg−1) 1 day before prime or boost, at prime or boost and 1 day after prime or boost, followed by serum and dLN collection at D56. b, WA1 spike-specific IgG at D56 in WA1mRNA or WA1mRNA+MA mice treated with saline (Sal; n = 5 per group), Ana (n = 7 per group) or V765 (n = 6 per group) as in a. c, Absolute numbers of CD19+CD95+GL7+ BGC cells (left) and CD4+CXCR5+CD278+ TFH cells (right) in the dLNs of mice as in b. d, Serum nAb titers against pseudovirus expressing the spike of WA1 (left), BA.5 (middle) or XBB.1.5 (right) in mice as in b. e, Absolute numbers of WA1 (left), BA.4/BA.5/BA.5.2 (middle) or XBB.1.5 (right) spike+ BGC cells quantified by flow cytometry in mice as in b. f, Schematic showing C57BL/6J wild-type mice immunized i.m. with saline, WA1mRNA or WA1mRNA+MA on D0 (prime) and on D14 (boost) and injected intravenously with IFNAR1 Ab+IFNγ Ab and the corresponding isotype (Iso), 200 μg per mouse, 1 day before prime or boost, at prime or boost and 3 days after prime or boost, followed by serum and dLN collection at D56. g, WA1 spike-specific IgG at D56 in WA1mRNA or WA1mRNA+MA mice treated with Iso (n = 5 per group) or IFNAR1 Ab+IFNγ Ab (n = 7 per group) as in f. h, Absolute numbers of BGC cells (left) and TFH cells (right) in the dLNs of mice as in g. i, Serum nAb titers against pseudovirus expressing the spike of WA1 (left), BA.5 (middle) or XBB.1.5 (right) on D56 in mice as in g. j, Absolute numbers of WA1 (left), BA.4/BA.5/BA.5.2 (middle) or XBB.1.5 (right) spike+ BGC cells quantified by flow cytometry in mice as in g. k, Percentage of CD19−CD38+CD138+ LLPCs as assessed by flow cytometry in the bone marrow of mice as in g. l, WA1 spike-specific IgG titer in BALB/c wild-type mice immunized with WA1mRNA or WA1mRNA+MA (n = 6 mice per group) and BALB/c Il1rn−/− mice immunized with WA1mRNA (n = 6) on D0 (prime) and on D14 (boost), followed by serum and dLN collection at D56. m, Absolute numbers of spike+ BGC cells of WA1 (left), BA.4/BA.5/BA.5.2 (middle) or XBB.1.5 (right) (n = 5 mice per group) quantified by flow cytometry in the dLNs of mice as in l. n, Serum nAbs against pseudoviruses expressing the spike protein of WA1 (left), BA.5 (middle) or XBB.1.5 (right) at D56 in mice as in l. Significance was determined using two-way ANOVA with Tukey’s multiple comparisons test (b−e,g−n). ‘n’ indicates the number of biologically independent samples. Box plots depict the median as the center line, the interquartile range as the box and the range with whiskers extending to the minimum and maximum values (b−e,g−n). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Ab, antibody; D, day; neut., neutralization; NS, not significant; WT, wild-type.
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