FIGURE 1.

Characterization and quantization of AMR‐derived EV. (A) Quantization of EV isolated from plasma of AMR patients was assessed by nanoparticle tracking analysis (Nanosight™ technology, NTA EV/ml) after isolation by ultracentrifugation. Kruskal–Wallis test (p = .031) revealed a significant difference among all groups (Healthy EV n = 5, for TX Ctrl, AAMR and CAMR n = 9). Dunn’s multiple comparisons post hoc test confirmed a significant increase in the number of EV in the AAMR group compared with the CAMR (p = .02). (B) Size distribution of plasma EV by nanoparticle tracking analysis. (C) Representative transmission electron microscopy showing EV size of a TX Ctrl patient. (D) Multiplex bead‐based flow cytometry analysis was performed by MACsplex exosome capture beads containing a cocktail of 39 different markers. Experiments were performed with EV from four patients for TX Ctrl, AAMR, and CAMR groups, graph indicated values after normalization of the raw median fluorescence intensity (MFI). Raw MFI was subtracted with the MFI of the negative/blank control used in the same run experiment to avoid nonspecific signals. Values below the corresponding control were indicated as negative and were not showed. (TX CTRL, AAMR, and CAMR n = 4, data are expressed as mean ± SD; *p value < .05, Kruskal–Wallis test.) [Color figure can be viewed at wileyonlinelibrary.com]