Fig. 7

CardAP-EVs attenuate anti-CD3 induced pro-inflammatory cytokine release only in the presence of CD14⁺ cells. CD3⁺ and CD14⁺ cells were isolated by MACS and cultured unstimulated for 48 h. Here, CD14⁺ cells were additionally treated with either unstimulated (EVs) or cytokine stimulated (EVs^(cyt)) EVs, PBS in equal volume of the EVs (PBS) or left untreated. After 2 days, CD14⁺ cells were co-cultured 1:5 with CD3⁺ cells and stimulated with anti-CD3. As a negative control, anti-CD3 stimulated T cells were treated with either CardAP-EV variant, PBS or left untreated. After 3 days, the supernatants were collected and cytokine concentrations were analysed by ELISA (IFNγ, active TGFβ) or Multiplex (IL-10, TNFα). Concentrations for all tested cytokines are presented for anti-CD3 stimulated monoculture of CD3⁺ cells (a) or co-cultures of CD3⁺ cells with CD14⁺ cells (b) as median with interquartile range (co-culture n = 6, five different CardAP donors, five different PBMC donors) (monoculture n = 8; four different CardAP donors, four different PBMC donors). Friedman Test with Dunn´s multiple comparison test: ***p < .001, **p < .01, *p < .05 or Wilcoxon matched-pairs signed rank test; ^###p < .001, ^##p < .01, ^#p < .05