Midline 1 kao posrednik proupalne signalizacije receptora Fas u mijeloidnim stanicama mišje koštane srži

master's thesis

• Title (english): Midline 1 as a mediator of infalmmatory signaling of Fas receptor in murine myeloid bone marrow cells
• Author: Sara Priselac
• Mentor: Nataša Kovačić (mentor)
• Committee member: Danka Grčević (predsjednik povjerenstva)
• Committee member: Ivan Banovac (član povjerenstva)
• Committee member: Nataša Kovačić (član povjerenstva)
• Granter: University of Zagreb; School of Medicine; (Department of Anatomy and Clinical Anatomy); Zagreb
• Defense date and country: 2023-09-19, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Basic Medical Sciences; Anatomy

Abstract

Mijeloidne stanice pridonose pokretanju i održavanju upalnog odgovora, što uz nedostatnu regulaciju rezultira oštećenjem tkiva koje prati mnoge autoimunosne bolesti. Sustav Fas/ligand Fas može ograničiti upalni odgovor apoptozom imunosnih stanica, a može ga i potaknuti slabo istraženim proupalnim mehanizmima. Nedavno smo u mišjem modelu artritisa opisali proupalnu signalnu os receptora Fas putem bjelančevine Mid 1 u mijeloidnim stanicama. Cilj rada bio je istražiti učinak neapoptotske signalizacije receptora Fas na izražaj gena Mid 1 i proupalnih citokina u upalno potaknutim stanicama koštane srži, te važnost Mid 1 u njihovoj upalnoj aktivaciji.

Stanice su inkubirane in vitro s oligomeriziranim i različitim dozama neoligomeriziranog agonističkog protutijela anti-Fas, sa i bez upalnog podražaja inaktiviranom bakterijom M. tuberculosis. Udio apoptotičnih stanica određen je protočnom citometrijom na temelju vezanja aneksina V. Izražaj gena proupalnih citokina IL-1β i TNF-α te gena Mid 1 određen je postupkom kvantitativne lančane reakcije polimeraze u stvarnom vremenu.

Rezultati su pokazali da oligomerizirano agonističko protutijelo djeluje isključivo apoptotski, a neoligomerizirano agonističko protutijelo anti-Fas izaziva apoptozu malog udjela mijeloidnih stanica te u upalno potaknutim stanicama dodatno potiče izražaj gena proupalnih citokina i Mid 1. Inaktivacija gena Mid 1 nije u potpunosti zakočila porast izražaja gena proupalnih citokina u upalno potaknutim stanicama koštane srži.

Abstract (english)

Myeloid cells initiate and support inflammation, which damages tissues in the course of autoimmune diseases. Fas/Fas ligand system hamper the inflammatory response via immune cell apoptosis, but can also enhance it via less understood proinflammatory mechanisms. We recently described a proinflammatory Fas-Mid 1 signaling axis in myeloid cells during murine arthritis. The aim of this study was to investigate the effect of non-apoptotic Fas signaling on the expression of Mid 1 and proinflammatory cytokine genes in murine bone marrow cells, and the role of Mid 1 in their inflammatory response.

Cells were treated in vitro with oligomerized and various concentrations of non-oligomerized agonistic anti-Fas antibody, in the presence or absence of inactivated M. tuberculosis. The proportion of apoptotic cells was determined by annexin V binding using flow cytometry. Gene expression of IL-1β, TNF-α, and Mid 1 was assessed by quantitative real-time polymerase chain reaction.

Results revealed a sole apoptotic effect of the oligomerized agonistic antibody, while the non-oligomerized antibody had a weak apoptotic effect on myeloid cells, but upregulated Mid 1 and proinflammatory cytokine genes in inflammatory stimulated cells. Inactivation of Mid 1 gene did not completely abolish the upregulation of proinflammatory cytokine genes in inflammatory stimulated bone marrow cells.

• Keywords: Fas; Mid 1; apoptoza; upala; mijeloidne stanice
• Keywords (english): Fas; Mid 1; apoptosis; inflammation; myeloid cells
• Language: croatian
• URN:NBN: urn:nbn:hr:105:925526
• Study programme: Title: Medicine; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: doktor/doktorica medicine (doktor/doktorica medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2023-10-30 11:03:13