Uvod: Oboljenja iz grupe pemfigusa - pemfigus vulgaris (PV) i pemfigus foliaceus (PF) su autiomunska bulozna oboljenja kože i sluznica. U patogenezi su od značaja autoantitela na dezmogleine i druge molekule koja dovode do akantolize i formiranja intraepidermalnog rascepa. Za aktivaciju autoreaktivnih T-ćelija u pemfigusu neophodno je prepoznavanja dezmogleinskih peptida i angažovanje CD28 molekula, koji je najznačajniji kostimulatorni receptor u aktivaciji naivnih T ćelija. CD28 molekul se vezuje za B7 familiju molekula kao što su CD80 i CD86. Pored toga, B7 molekuli mogu da aktiviraju inhibitorni receptor na T-ćelijama, CTLA-4, koji kontroliše mehanizam održavanja tolerancije na sopstvene peptide. Na značaj genetskih faktora u patogenezi pemfigusa ukazuju prethodna istraživanja kojima su definisani određeni genetski faktori kao što su i polimorfizmi pojedinačnih nukleotida u genima molekula od značaja za patogenezu pemfigusa.
Ciljevi istraživanja: Odrediti distribuciju alela u genima za molekule CD86, CTLA-4, TNF i IL-10 kod pacijenata sa PV i PF, kao i kod zdravih osoba (dobrovoljni davaoci krvi) u Srbiji, ispitati da li je neki od ispitivanih polimorfizama faktor rizika za nastanak oboljenja (PV i PF) i uporediti ih sa podacima iz literature koji su dobijeni ispitivanjem istih polimorfizama kod geografski i etnički različitih populacija.
Pacijenti i metode: Istraživanje je obuhvatilo 61 bolesnika sa pemfigusom od kojih je PV imalo 48, a PF 13 bolesnika i 486 zdravih osoba-kontrola (dobrovoljni davaoci krvi). Dijagnoza pacijenata sa pemfigusom postavljena je na osnovu kliničke slike i testa direktne imunofluorescencije (DIF test). Za detekciju i analizu polimorfizama gena za CD86, CTLA-4, TNF i IL-10 korišćene su komercijalne optimizovane smeše specifičnih oligonukleotida i TaqMan proba obeleženih FAM i VIC fluorohromima i to: CD86 rs1129055, CTLA4 rs733618 i rs5742909, TNF rs1800629 i rs361525 i IL-10 rs1800896 i rs1800871 (PE, Applied Biosystems). Amplifikacija fragmenata DNK Real-time PCR metodom vršena je pomoću navedenih smeša komercijalnih oligonukleotida i
Maxima™ Probe qPCR 2X Master Mix, (Fermentas) uzimajući u obzir preporuke proizvođača reagenasa...Introduction: Pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are rare autoimmune blistering diseases with presumed T cell dependent pathology. Activation of naïve T cells is dependent on antigen recognition, subsequent signaling through the T cell receptor complex (signal 1), and various other interactions of T cells with antigen presenting cells that may be collectively designated as signal 2, which is unconditionally required for T cell activation both in response to infection and to autoantigens. Among the best described interactions contributing to signal 2 are those mediated by B7 family molecules such as CD80 and CD86 with their ligands, CD28, providing activation signals, and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), conferring inhibition. Single nucleotide polymorphisms (SNPs) within genes encoding those molecules may alter the signaling process. It is not known whether functional genetic polymorphisms within genes encoding the aforementioned proteins may increase risk for developing PV and PF and, if so, whether they might serve as biomarkers for susceptibility to these diseases. Aims of the investigation: To examine functional single nucleotide polymorphisms within CD86 (rs1129055), CTLA4 (rs733618 and rs5742909), TNF (rs1800629 andrs361525), and IL-10 (rs1800896 and rs1800871) genes in 61 pemphigus patients and 486 healthy controls, and to compare results in Serbian pemphigus patients and healthy controls with results obtained in different populations groups. Patients and methods: Our study included 61 pemphigus patients, among them 48 with PV and 13 with PF, treated at the Dermatovenereology Clinics of the Clinical Centre of Serbia. Patients were stratified according to clinical presentation of the disease as a PV or PF patients. Diagnosis in all patients was established by merging clinical findings, histopathologic analysis, and the results of direct immunofluorescence tests. Blood samples from 486 healthy blood donors were obtained from the National Blood
Transfusion Institute..
