Proteomic analysis of extracellular vesicles from Trypanosoma cruzi trypomastigotes revealed that contain a large amount of proteins involved in pathogenesis.

CAEIRO, L; TEKIEL, V

Buenos Aires

Congreso; Reunión Conjunta de Sociedades de Biociencias.; 2017

SAIB-SAP-SAIC-SAi, etc

PROTEOMIC ANALYSIS OF EXTRACELLULAR VESICLES FROM Trypanosoma cruzi TRYPOMASTIGOTES REVEALED THAT CONTAIN A LARGE AMOUNT OF PROTEINS INVOLVED IN PATHOGENESISThe trypomastigote is the circulating bloodstream and non-replicative stage of Trypanosoma cruzi, the causative agent of Chagas disease. Many of the surface proteins of the trypomastigote, involved in cell invasion and/or in the infection process, are also secreted. We have previously found that trypomastigotes secrete into extracellular vesicles (EVs) virulence factors that should be hidden from the immune system. Data currently available from exoproteomes were derived from total secreted material (free and vesicle-contained, Y strain) or from a mixture of EVs from parasites and host cells (Tulahuen strain). Here we present a proteomic analysis of trypomastigote EVs (reference CL Brener strain), to search for putative virulence factors secreted into trypomastigotes EVs. To isolate EVs, conditioned media from cell-derived trypomastigotes were sequentially ultracentrifugated to obtain, after 2 and 16 hs, EVs fractions V2 and V16, respectively. Both fractions were analyzed in a Q Exactive HESI-Orbitrap coupled to a nano HPLC Easy-nLC 1000. MS/MS data were used to search T. cruzi database (Tritrypdb.org, v.30). Each EV subpopulation presented a differential set of major proteins (V2: n=270; V16: n=228), and a minor core of 142 common proteins. GO Analysis (UniprotKB) revealed that V2 and V16 had mostly proteins involved in pathogenesis (37,8%; 56,7%) and cellular process (22,1%; 12%). We also found known virulence factors like KMP-11, several members of multigene families (TS, TcTASV) and immune system modulators (i.e. TcHMGB), some of them were among the common core like TcTASV-C, TS group I and TolT. This data demonstrate that EVs contain a large number of key proteins involved in multiple regulatory and pathogenic processes. Characterization of trypomastigote EVs cargo may be helpful to identify novel vaccine or drug targets for the control of Chagas disease.Supported by PICT 2014-1151; PIP-2015-186.Keywords: Trypanosoma cruzi, trypomastigote, extracellular vesicles, proteome, pathogenesis