Extracellular Vesicles in Allergic Rhinitis and Asthma and Laboratory Possibilities for Their Assessment

Abstract

Currently, extracellular vesicles (EVs) have been implicated in the etiopathogenesis of many diseases, including lung disorders, with the possibility of diagnostic and therapeutic applications. The analysis of EV in respiratory tract diseases faces many obstacles, including material collection from airways, standardization of isolation techniques, detection methods, the analysis of their content, etc. This review focuses on the role of extracellular vesicles in the pathogenesis of atopic respiratory diseases, especially asthma, with a special focus on their clinical applicability as a diagnostic tool. We also summarize available laboratory techniques that enable the detection of EVs in various biological materials, with particular emphasis on flow cytometry. The opportunities and limitations of detecting EV in bronchoalveolar lavage fluid (BALF) were also described.

Keywords: allergy; asthma; extracellular vesicles; flow cytometry; laboratory techniques.

Figure 1

Example of dot plots and histogram of megamix-Plus SSC polystyrene beads using DIVa 8.0.1 software and FACSCantoII flow cytometer (Becton, Dickinson and Company) (own results, unpublished).

Figure 2

Example of isotypic control and platelets-, endothelial-, leukocytes- EVs with anti-CD42b-APC, anti-CD63-PE, anti-CD45-PE-Cy7 antibodies respectively in Tru-count tube with LOW Tru-count control beads using DIVa 8.0.1 software and FACSCantoII flow cytometer (Becton, Dickinson and Company) (own results, unpublished). As an internal control of the assessments of the absolute number of particles, manufacturer controls were used (LOW control beads were used because the number of detected particles was small).

Figure 2

Example of isotypic control and platelets-, endothelial-, leukocytes- EVs with anti-CD42b-APC, anti-CD63-PE, anti-CD45-PE-Cy7 antibodies respectively in Tru-count tube with LOW Tru-count control beads using DIVa 8.0.1 software and FACSCantoII flow cytometer (Becton, Dickinson and Company) (own results, unpublished). As an internal control of the assessments of the absolute number of particles, manufacturer controls were used (LOW control beads were used because the number of detected particles was small).