1. Impact of ONX 0914 on abundance and dissemination of monocytes/macrophages during CVB3 infection
Impact of ONX 0914 on abundance and dissemination of monocytes/macrophages during CVB3 infection A, BNaive A/J mice were treated solely with ONX 0914 or vehicle daily for 9 days (A) (dark blue vs. gray bars) or with a single shot (B) (18 h; light blue vs. white bars/dots). Subsequently, blood and spleen (A) were isolated and analyzed using flow cytometry. Plotted are means of total numbers of the indicated myeloid (sub‐)populations + SEM. Unpaired t‐tests (+ Welch correction) were performed (one experiment; vehicle n = 4, ONX n = 6). Splenocytes from ONX 0914‐ or vehicle‐treated mice were first labeled with fluorochrome‐conjugated antibodies against the surface markers CD11b and F4/80 (B). To visualize and monitor phagocytosis, cells were incubated with fluorophore‐labeled particles for the indicated time and analyzed using flow cytometry. After gating CD11b+/F4/80+ macrophages, amount of phagocytosed labeled particles was determined by MFI per 105 cells (means + SEM) (one representative experiment out of two is shown; vehicle n = 4, ONX n = 4; two‐way ANOVA followed by Bonferroni's multiple comparisons test). P‐values are indicated in each graph.C, DA/J mice were infected with CVB3. ONX 0914 or vehicle treatment was carried out daily, starting one day prior to virus inoculation. 3 (C) and 8 days (D) after infection, mice were sacrificed, spleen was isolated (1 (D) or 4 (C) separate experiments were carried out, respectively; d3: vehicle n = 9–12, ONX n = 10–12; d8: E3 vehicle n = 3, ONX n = 8), and the number of different myeloid cell types was determined by flow cytometry. A detailed description of the gating strategy for identification of the different immune cell type is provided in Fig EV2. Means + SEM are shown, and unpaired t‐tests (+ Welch correction) were performed. P‐values are indicated in each graph.
Nadine Althof et al. EMBO Mol Med. 2018;emmm
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