E-resources
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Junec, Franjo; Jurenec, Franjo
2016Web Resource
Antitimocitni globulini su poliklona protutijela koja dovode do smanjenja broja T stanica. Često se koriste za imunosupresiju primatelja tijekom transplantacije bubrega. Timoglobulin su IgG protutijela dobivena imunizacijom zečeva timocitima ljudskog fetusa. Timoglobulin dovodi do smanjenja broja T-limfocita putem razgradnje stanica mehanizmom ovisnim o komplementu te apoptozom. Da bismo dokazali direktan učinak timoglobulina na stanice, koristili smo ljudske embrionalne stanice porijeklom iz bubrega (HEK-293) u staničnoj kulturi. Mjerili smo membranski potencijal stanice metodom prikovanih potencijala. Depolarizacijski učinak timoglobulina na membranski potencijal HEK-293 stanica je ovisan o koncentraciji timoglobulina te o početnom membranskom potencijalu stanica. Učinak timoglobulina u hipoksično-reoksigenacijskih uvjetima utvrđen je određivanjem stanične smrtnosti te stanične migracije testom grebanja. Timoglobulin je smanjio staničnu smrtnost u hipoksijskim uvjetima nakon kojih je slijedila reoksigenacija. Ovo je prvo istraživanje koje pokazuje direktan učinak timoglobulina na HEK-293 stanice. Rezultati ovog istraživanja potiču nova istraživanja o djelovanju timoglobulina na stanice transplantiranih bubrega. Antithymocyte globulines are polyclonal T cell-depleting immunoglobulins used in induction of immunosuppression in kidney transplant recipients. Thymoglobuline, is purified rabbit IgG, obtained by immunization of rabbits with fetal human thymi, which depletes T lymphocytes by complement-dependent lysis and apoptosis. To determine possible direct effects of Thymoglobuline on the cells, we used Human Embryonic Kidney cell line (HEK-293) in culture. We measured membrane potential of the cells using slow whole patch clamp technique. Depolarizations of HEK-293 cells caused by Thymoglobuline were concentration- and membrane potential- dependent, showing direct effect of Thymoglobuline on the HEK-293 cells. The effects of Thymoglobuline in hypoxia/reoxigenation were detected by calculating cell death and determining the cell migration using scratch test. Thymoglobuline prevented the cell death induced by hypoxia and reoxigenation conditions. This is first study showing direct effect of thymoglobulin on HEK-293 cells. The results of this study encourage the research on epithelial cells in transplanted kidneys.
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