A sejtek közti kommunikáció újonnan azonosított mikrovezikulum-útjának vizsgálata = Analysis of cell-derived microvesicles that represent novel players in intercellular communication

Munkank soran az extracellularis vezikulak izolalasanak es detektalasanak szamos meghatarozo preanalitikai es analitikai parameterere hivtuk fel a figyelmet. Elsőkent mutattunk ra, hogy a mikrovezikulak es a feherje-aggregatumok biofizikai parameterei jelentős mertekben atfednek, es ez zavarhatja a mikrovezikulak mereset. Kidolgoztuk annak modszeret, hogy egyazon biologiai forrasbol szarmazo kulonboző vezikula populaciokat parhuzamosan, nagy mennyisegben, intakt formaban tudjunk izolalni. Osszehasonlito proteomikai elemzest vegeztunk thymus eredetű apoptotikus testek es mikrovezikulak eseteben. Szamos T sejt jelatvitelben es immunfolyamatokban szerepet jatszo feherjet es autoantigent azonositottunk. Igazoltuk, hogy T sejt eredetű citokinek es extracellularis vezikulak egyuttes hatasat monocitak genexpressziojara. Igazoltuk, hogy a mikrovezikulak onallo ionhaztartassal rendelkeznek. Kimutattuk, hogy thymocyta exoszomak nem tartalmaznak riboszomalis RNS-eket, azonban feldusulnak bennuk kis RNS-ek (pl. bizonyos miRNS-ek). Polymyositises betegekben emelkedett keringő mikrovezikula szamot mutattunk ki, mely korrelalt a betegseg bizonyos klinikai parametereivel. Vegul elsőkent igazoltuk, hogy egeszseges T sejt eredetű mikrovezikulak CD62P-CD161 kolcsonhatas reven specifikusan kotődnek monocitak felszinehez. | In our work we drove attention to several pre-analytical and analytical parameters affecting isolation and detection of work extracellular vesicles. We were the first to describe that microvesicles share biophysical parameters with protein aggregates which may confound microvesicle assessment by flow cytometry. We developed protocols for the isolation of large amounts of intact vesicle types secreted simultaneously by the same biological source. We carried comparative proteomic analysis of murine thymus derived apoptotic bodies and microvesicles. We identified large number of proteins involved in T cell signaling or immune functions as well as autoantigens within these structures. We provided evidence fro crosstalk between T cell derived extracellular vesicles and cytokines on the gene expression of monocytes. We have shown that microvesicles possess autonomous ion homeostasis. We found that thymocyte derived exosomes lacked the 18S and 28S ribosomal RNA molecules, while they were enriched in small RNA species (e.g. certain miRNAs). We described that patients with polyomyelitis were characterized by elevated levels of circulating microvesicle. Monocyte- and B cell-derived microvesicle numbers correlated with certain clinical parameters of the diseases. Finally, for the first time we showed that HLA-G+, trophoblast-derived microvesicles isolated from healthy pregnant blood plasma samples, bound specifically to T cells via CD62P-CD161 interaction, and induced STAT3 phosphorylation.