Vpliv veziklov mikro RNA na eksocitozo citotoksičnih limfocitov T pri diabetesu tipa 1

Horvat, Monika (author), Ihan, Alojz (mentor)

Diabetes tipa 1 je avtoimunska bolezen, ki se razvije kot posledica uničenja celic β,ki ga vršijo citotoksični limfociti T. Natančen razvoj bolezni je danes še vedno neznan. Namen magistrske naloge je bil raziskati vpliv veziklov z mikro RNA (veziklov miRNA) na eksocitozo citotoksičnih limfocitov T pri bolnikih z diabetesom tipa 1. Dokazane miRNA, ki sprožijo eksocitozo bi lahko uporabili kot biomarker, ki bi napovedoval razvoj diabetesa. Izvedli smo transfekcijo perifernih mononuklearnih celic z vezikli miRNA s transfekcijskim reagentom DOTAP. S testom citotoksičnosti CD107a smo dokazovali vpliv veziklov miRNA. V študijo je bilo vključenih 8 bolnikov z diabetesom tipa 1 in 8 zdravih posameznikov. Ugotovili smo, da miR-375 poveča vpliv na eksocitozo citotoksičnih limfocitov pri obeh preiskovanih skupinah. Dokazali smo, da ni razlike v vplivu miR-375 na celice bolnikov in zdravih posameznikov. Na treh bolnikih in treh zdravih posameznikih, smo dokazovali vpliv miRNA na sekrecijo TNF-α, IL-1β in IL-6. Citokine smo detektirali s komercialnim kompletom za detekcijo zunajceličnih citokinov. Ugotovili smo, da ima miR-375 pri posamezni preiskovani skupini večji vpliv na sekrecijo preiskanih citokinov kot RNA41; vendar ni značilne razlike med skupino bolnikov in zdravih posameznikov.

• Language: Slovenian
• Keywords: diabetes tipa 1, mikro RNA, eksocitoza, citotoksičnost, test citotoksičnosti CD107a, limfociti
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [M. Horvat]
• Year: 2018
• PID: 20.500.12556/RUL-102664
• UDC: 577.27.083.36:616.379-008.64
• COBISS.SI-ID: 4937080
• Publication date in RUL: 06.09.2018

Secondary language

• Language: English
• Title: The effect of microRNA vesicles on exocytosis of cytotoxic lymphocytes T in type 1 Diabetes

Abstract

Type 1 Diabetes is an autoimmune disease, in which insulin secreting β cells are destroyed by cytotoxic lymphocytes T. Clear onset of disease is still unknown. The aim of the Master thesis is to investigate the effect of microRNA carried by vesicles on exocytosis of cytotoxic lymphocytes in type 1 Diabetes in patients and to compare the effect to the healthy individuals. Possible miRNA could serve as a biomarker in development of the Diabetes. We transfected isolated peripheral blood mononuclear cells with DOTAP-miRNA vesicles. The effect of vesicles microRNA on exocytosis of cytotoxic lymphocytes was measured with a test of cytotoxicity CD107a. Eight healthy individuals and eight patients with diabetes were included in our study. We found, that miR-375 has a higher effect on exocytosis of cytotoxic lymphocytes in both investigated groups. We proved, that there is no difference in effect of miR-375 on cells between patient and healthy individuals. We measured the secretion of cytokines TNF-α, IL-1β and IL-6 with a commercial kit for detection of extracellular cytokines in three healthy individuals and three patients. We wanted to show the effect of vesicle miRNA on secretion of cytokines. We found, that miR-375 has a higher effect on secretion of cytokines in comparison with RNA41 in both investigated groups, but the difference in effect of miR-375 between patients and healthy individuals was not statistically significant.

• Keywords: type 1 Diabetes, microRNA, exocytosys, cytotoxicity, test of cytotoxicity CD107a, lymphocytes