Characterization of Extracellular Vesicles Secreted by a Clinical Isolate of Naegleria fowleri and Identification of Immunogenic Components within Their Protein Cargo
Metadatos
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MDPI
Materia
Extracellular vesicles Naegleria fowleri Characterization Immunogenic Proteases Proteome
Fecha
2022-06-29Referencia bibliográfica
Retana Moreira, L.; Steller Espinoza, M.F.; Chacón, Camacho, N.; Cornet-Gomez, A.; Sáenz-Arce, G.; Osuna, A.; Lomonte, B.; Abrahams Sandí, E. Characterization of Extracellular Vesicles Secreted by a Clinical Isolate of Naegleria fowleri and Identification of Immunogenic Components within Their Protein Cargo. Biology 2022, 11, 983. [https://doi.org/10.3390/biology11070983]
Patrocinador
Vicerrectoría de Investigación” of the Universidad de Costa Rica by supporting the research projects C-1061: “Caracterización de antígenos de excreción/ secreción y antígenos somáticos en amebas de vida libre mediante empleo de anticuerpos policlonales producidos en roedores” and C-2600: “Secreción de vesículas extracelulares por Naegleria fowleri y evaluación de su potencial rol inmunomodulador en un modelo in vitro”Resumen
Extracellular vesicles (EVs) are small lipid vesicles released by both prokaryotic and
eukaryotic cells, involved in intercellular communication, immunomodulation and pathogenesis.
In this study, we performed a characterization of the EVs produced by trophozoites of a clinical
isolate of the free-living amoeba Naegleria fowleri (N. fowleri). Size distribution, zeta potential, protein
profile and protease activity were analyzed. Under our incubation conditions, EVs of different sizes
were observed, with a predominant population ranging from 206 to 227 nm. SDS-PAGE revealed
protein bands of 25 to 260 KDa. The presence of antigenic proteins was confirmed byWestern blot,
which evidenced strongest recognition by rat polyclonal antibodies raised against N. fowleri in the
region close to 80 KDa and included peptidases, as revealed by zymography. Proteins in selected
immunorecognized bands were further identified using nano-ESI-MS/MS. A preliminary proteomic
profile of the EVs identified at least 184 proteins as part of the vesicles’ cargo. Protease activity assays,
in combination with the use of inhibitors, revealed the predominance of serine proteases. The present
characterization uncovers the complexity of EVs produced by N. fowleri, suggesting their potential
relevance in the release of virulence factors involved in pathogenicity. Owing to their cargo’s diversity,
further research on EVs could reveal new therapeutic targets or biomarkers for developing rapid and
accurate diagnostic tools for lethal infections such as the one caused by this amoeba.