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. 2021 Jul;76(7):2276-2279.
doi: 10.1111/all.14792. Epub 2021 Mar 20.

Proteomic profile of extracellular vesicles in anaphylaxis and their role in vascular permeability

Emilio Nuñez-Borque  1 , Sergio Fernandez-Bravo  1 , Carlos Pastor-Vargas  1   2   3 , Gloria Alvarez-Llamas  4   5 , Maria Dolores Gutierrez-Blazquez  6 , Ebrahim Alwashali  6 , Jose Julio Laguna  3   7   8 , Javier Dionicio  3   7 , Diana Betancor  1 , Victoria Villalobos  1 , Jaime Tome-Amat  3   9 , Javier Cuesta-Herranz  1   3 , Alberto Benito-Martin  10   11 , Vanesa Esteban  1   3   8
Affiliations

Proteomic profile of extracellular vesicles in anaphylaxis and their role in vascular permeability

Emilio Nuñez-Borque et al. Allergy. 2021 Jul.
No abstract available

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Conflict of interest statement

There are no conflicts of interest.

Figures

FIGURE 1
FIGURE 1
Characterization, proteomic profiling, and differential analysis of AnEVs and BEVs. (A) Transmission electron microscopy images from representative circulating BEVs and AnEVs. Scale bar: 200 nm. Images depict representative EVs from both samples. (B) Seven different EV preparations from each experimental condition were analyzed by NTA. Representative NTA histograms showed average particle sizes of 200 nm for BEVs and AnEvs. (C) AnEVs were characterized by Western blot. Panels show immunoblots from three representative patients. Bona fide EV markers, such as CD63, TSG101, Syntenin‐1, and CD9 were detected. (D) Venn diagram of brute data obtained by mass spectrometry‐based quantitative proteomics representing the intersection between detected proteins in BEVs and AnEVs. (E) Volcano plot showing all proteins identified. Statistically significant differences (p > .05) in the AnEVs (right side) and in the BEVs (left side) appear in blue. Access number (UniProt) for proteins of interest (CDC42, Ficolin‐2, S100A9) are shown. Principal component analysis (F) and unsupervised hierarchical clustering (G) of plasma‐derived EVs
FIGURE 2
FIGURE 2
Functional protein association networks in AnEVs. (A) CDC42, S100A9, and Ficolin‐2 are increased in AnEVs. The graphics represent the abundance protein levels expressed as the quantified ratio (AnEVs/BEVs and BEVs/BEVs) of a larger group of anaphylactic plasma paired samples. CDC42 (*p = .0137, n = 15, MW=21 KDa), S100A9 (**p = .0017, n = 20, MW = 14 KDa), Ficolin‐2 (**p = .0092, n = 26, MW = 34 KDa). (B) Classification of the main cellular localization and function of the protein panel based on the UniProt database. (C) Top Canonical Pathways obtained by IPA and symbols of the molecules. (D–E) HMVEC‐Ls were incubated with 100 µg/ml of purified BEVs and AnEVs. The images reveal PKH67‐EVs around the nuclei (DAPI) and cytoskeletal fibers (F‐actin) from a representative paired EVs patient sample from 4 performed (D). (E) The graphic shows the change in TEER measurements after the addition of BEVs and AnEVs (n = 16 patients). Two‐way ANOVA followed by the Bonferroni test was performed: **p = .0033, ****p ≤ .0001 versus Vehicle (EBM + PBS); #p = .0285 versus BEVs)
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