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Extracellular vesicles isolated from Toxoplasma gondii induce host immune response
Author(s) -
Silva Valeria Oliveira,
Maia Marta Marques,
Torrecilhas Ana Claudia,
Taniwaki Noemi Nosomi,
Namiyama Gislene Mitsue,
Oliveira Katia Cristina,
Ribeiro Kleber Silva,
Toledo Maytê dos Santos,
Xander Patricia,
PereiraChioccola Vera Lucia
Publication year - 2018
Publication title -
parasite immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.795
H-Index - 75
eISSN - 1365-3024
pISSN - 0141-9838
DOI - 10.1111/pim.12571
Subject(s) - toxoplasma gondii , microvesicles , biology , immune system , toxoplasmosis , extracellular , nanoparticle tracking analysis , vesicle , rna , secretion , extracellular vesicle , microbiology and biotechnology , virology , antibody , immunology , biochemistry , membrane , microrna , gene
Summary This study established a protocol to purify Toxoplasma gondii tachyzoite microvesicles and exosomes, called as extracellular vesicles ( EV s). In addition, the investigations were conducted to determine the kinetic of EV release by tachyzoites and whether EV proteins are able to modulate the host immune response. The particle size and concentration released by tachyzoites in culture medium at different incubation‐period were characterized by nanoparticle tracking analysis. Tachyzoites (1 × 10 6 ) released around 4.37 ± 0.81 × 10 8 EV s/ mL /h, with size varying between 138.2 and 171.9 nm. EV s released into the medium were purified by gel‐exclusion chromatography and screened by ELISA , using a pool of human positive sera for toxoplasmosis. EV ‐fractions contained high concentration of proteins, and EV s were analyzed by scanning and transmission electron microscopies. Tachyzoites released EV s into the culture medium throughout all membrane surface, and these vesicles contain small RNA s/mi RNA . Pooled sera from chronically infected human or mice (infected with 2 different T. gondii strains) recognized distinct EV electrophoretic patterns in immunoblotting. T. gondii EV s significantly induced IL ‐10, TNF ‐α and iNOS in murine macrophages. In conclusion, this study shows that T. gondii secrete/excrete EV s (microvesicles and exosomes) contain mi RNA and they were immunologically recognized by host immune response.