Vpliv visoko avidnih protiteles proti ß2-glikoproteinu I na izločanje vnetnih molekul iz človeških endotelijskih celic koronarne arterije : [magistrski študij laboratorijska biomedicina]

Koželj, Mateja (author), Božič, Borut (mentor), Sodin-Šemrl, Snežna (comentor)

Antifosfolipidni sindrom je avtoimunska bolezen, opredeljena s prisotnostjo antifosfolipidnih protiteles v plazmi bolnikov z venskimi ali arterijskimi trombozami in/ali ponavljajočimi zapleti v nosečnosti. Bolniki z antifosfolipidnim sindromom imajo večje tveganje za razvoj ateroskleroze, srčno žilne bolezni pa danes prištevamo med vodilne vzroke smrti po svetu. Naš namen je bil preveriti vpliv visoko avidnih protitelesa proti β2-glikoproteinu I na izločanje vnetnih molekul iz človeških endotelijskih celic koronarne arterije, gojenih in vitro. Protitelesa proti β2-glikoproteinu I, izolirana iz vzorca imunoadsorpcije bolnika z antifosfolipidnim sindromom, smo primerjali s protitelesno frakcijo, izolirano iz seruma zdravih krvodajalcev. Mirujoče in predhodno spodbujene celice z akutno-faznim proteinom serumskim amiloidom A smo izpostavili obema vzorcema protiteles. Po dvajset urni inkubaciji smo aktivacijo endotelijskih celic kvantitativno vrednotili z določanjem koncentracije citokinov, kemokinov in adhezivnih molekul na proteinskem nivoju z uporabo specifične encimsko-imunske metode. Človeške endotelijske celice koronarne arterije, spodbujene s protitelesi proti β2- glikoproteinu I bolnika z antifosfolipidnim sindromom, so v primerjavi s protitelesi razreda G zdravih krvodajalcev povečano izločale monocitni kemotaktični protein-1, koncentracija kemokina pa se je izrazito povišala po predhodni spodbuditvi celic s serumskim amiloidom A. Dodatek bolnikovih protiteles je povzročil tudi povišanje izločene koncentracije interlevkina-8 in interlevkina-6, pri čemer je koncentracija interlevkina-6 naraščala sorazmerno s koncentracijo predhodno dodanega serumskega amiloida A. Po dodajanju obeh vzorcev protiteles smo izmerili primerljive koncentracije medcelične adhezivne molekule-1. Pri izločanju žilnocelične adhezivne molekule-1 pa je dodatek protiteles proti β2-glikoproteinu povzročil zaviralni učinek, kar pa se ni ponovilo na celicah predhodno spodbujenih s serumskim amiloidom A. Ugotovili smo, da visoko avidna protitelesa proti β2-glikoproteinu I spodbujajo izločanje kemokinov, citokinov in adhezivnih molekul ter zavirajo izločanje žilnocelične adhezivne molekule-1. Učinek je intenzivnejši ob predhodno prisotnem vnetnem procesu. Visoko avidna protitelesa proti β2-glikoproteinu I preko spodbuditve izločanja vnetnih molekul vplivajo na razvoj ateroskleroze pri bolnikih z antifosfolipidnim sindromom.

• Language: Slovenian
• Keywords: antifosfolipidni sindrom ß2-glikoproteini ateroskleroza vnetje protitelesa izolacija
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: FFA - Faculty of Pharmacy
• Place of publishing: Ljubljana
• Publisher: [M. Koželj]
• Year: 2012
• Number of pages: VII, 59 f.
• PID: 20.500.12556/RUL-121144
• UDC: 616-097(043.3)
• COBISS.SI-ID: 3242097
• Publication date in RUL: 30.09.2020

KOŽELJ, Mateja, 2012, Vpliv visoko avidnih protiteles proti ß2-glikoproteinu I na izločanje vnetnih molekul iz človeških endotelijskih celic koronarne arterije : [magistrski študij laboratorijska biomedicina] [online]. Master’s thesis. Ljubljana : M. Koželj. [Accessed 27 March 2025]. Retrieved from: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=eng&id=121144

Secondary language

• Language: English
• Title: Effects of high avidity anti ß2-glycoprotein I antibodies on expression of inflammatory molecules from human coronary artery endothelial cells

Abstract

Antiphospholipid syndrome is an autoimmune disease defined by the presence of antiphospholipid antibodies in the plasma of patients with venous or arterial thrombosis and/or recurrent pregnancy complications. Patients with antiphospholipid syndrome suffer from increased risk of atherosclerosis and consequentially, cardiovascular diseases are now counted among the leading causes of death worldwide. Our aim was to examine the effect of high avidity antibodies to β2-glycoprotein I on the secretion of inflammatory molecules from human coronary artery endothelial cells cultured in vitro. Antibodies to β2-glycoprotein I were isolated from the imunoadsorption sample of a patient with antiphospholipid syndrome. Their activities were compared to the activities of an antibody fraction isolated from healthy blood donors. Resting and pre-stimulated cells with acute-phase protein Serum Amiloid A were exposed to the indicated samples of antibodies. After twenty hours, activation of endothelial cells was quantitatively evaluated by determining the released concentrations chemokines, cytokines and adhesion molecules using specific enzyme-linked immunosorbent assay. Human coronary artery endothelial cells that were stimulated with antibodies to β2- glycoprotein I from the patient with antiphospholipid syndrome were increasingly excreting Monocyte chemotactic protein-1 as compared with antibodies of class G of healthy blood donors. Also, the concentration of this chemokine was markedly increased by prior stimulation of the cells with Serum Amiloid A. Addition of patient antibodies led to higher concentrations of both Interleukin-8 and Interleukin-6, while only the concentration of Interleukin-6 increased in proportion to the applied concentrations of added Serum Amyloid A. Following the addition of the indicated samples of antibodies we have measured comparable concentrations of the Intercellular adhesion molecule-1. In regard to the release of Vascular cell adhesion molecule-1, the addition of anti-β2- glycoprotein antibodies from the patient, has caused a dampening effect, which was not observed when cells were pre-stimulated with Serum Amiloid A. We have found that high avidity β2-glycoprotein I antibodies stimulate secretion of chemokines, cytokines and adhesive molecules and inhibit the secretion of vascular cell adhesion molecule-1. The effect is more intense when the inflammatory process already existed. Antibodies to β2-glycoprotein I influence the development of atherosclerosis in patients with antiphospholipid syndrome via induced secretion of inflammatory molecules.