Fig. 3

Different protein coronae affect RSV infectivity and moDC activation. a, b RSV virions produced under serum-free conditions were pre-coated with different coronae via pre-incubation with different biological fluids prior to addition to HEp-2 cells in serum-free medium (diluted 10×) at an MOI of 1. The frequencies of GFP⁺ cells were quantified by flow cytometry 72 h post infection. Gating strategy and uninfected control is shown in Supplementary Fig. 5. a Representative dot blot graphs demonstrating GFP⁺ cell frequency from different corona conditions. b Flow cytometry quantification of the GFP⁺ cells presented as fold increase over RSV treatment in serum-free conditions (no corona). Data are presented as mean ± SEM of six replicates from two separate experiments. Significant differences in comparison with RSV (no corona) were assessed by non-parametric Kruskal–Wallis unpaired test followed by Mann–Whitney test. P-value: **P ≤ 0.01. c–f RSV virions produced under serum-free conditions were pre-coated with different coronae prior to the addition to primary moDCs in serum-free conditions at MOI of 20. CD86 expression and frequencies of GFP⁺ cells were quantified by flow cytometry 72 h post infection. c Representative histograms of CD86 expression in moDCs are shown. d CD86 mean fluorescence intensity (MFI) is shown as mean  ±SEM for six to eight individual donors from four separate experiments. Sucrose from the sucrose cushion used for RSV harvesting was also used as a control. e The frequency of GFP⁺ cells is shown as mean  ±SEM for six to eight individual donors from four separate experiments. Significant differences in comparison with RSV (no corona) were assessed by non-parametric Kruskal–Wallis unpaired test followed by Mann–Whitney test. P-value: **P ≤ 0.01 and *P ≤ 0.05. f Representative dot blot graphs demonstrating GFP⁺ moDCs frequency from different corona conditions