| Abstract | Cilj istraživanja: Cilj je istraživanja bio utvrditi fenotip i funkcionalni status sluznicama pridruženih invarijantnih T (MAIT) limfocita i gamma delta (γδ) T stanica u psorijazi, proučavajući njihove udjele u perifernoj krvi zajedno sa serumskim razinama IL-17, IL-23, IL- 18, CCL20 i CCL27 i transkripcijskim profilima gena u kontroli kemokinskih receptora (IL18R, CCR6, CCR10, SELPLG) i transkripcijskih čimbenika koji vjerojatno upravljaju razvojem urođenog (ZBTB16), citotoksičnog/tip 1 (RUNX3, EOMES, TBX21) i tip 17 (RORC) odgovora u sortiranim MAIT i γδ T stanicama zdravih i oboljelih ispitanika. Nacrt studije: opažajno istraživanje parova. Ispitanici: Istraživanje je obuhvatilo 21-og punoljetnog ispitanika oboljelog od vulgarne psorijaze (PV) i 22 zdrave, po dobi i spolu podudarne, nesrodne osobe koje su činile kontrolnu skupinu. Materijali i metode: Demografski, kliničko– antropometrijski, biokemijski te podatci postojanja infektivnih biljega prikupljeni su zajedno s uzorcima krvi i seruma. Težina kliničke slike određena je s pomoću PASI ljestvice i DLQI upitnika. Imunofenotipizacija MAIT i γδ T stanica učinjena je metodom višeparametrijske protočne citometrije, uključujući specifično obilježavanje MAIT stanica s MR1-5-OP-RU tetramerom. Luminex tehnologija korištena je za mjerenje serumskih razina citokina i kemokina, a ciljani qRT-PCR učinjen je u svrhu analize transkripcijskih profila sortiranih γδ i MAIT stanica Rezultati imunofenotipizacijskih i genotipizacijskih metoda analizirani su u odnosu na biokemijske, serološke, demografske i kliničke značajke ispitanika. Rezultati: Sastav perifernih MAIT i γδ T stanica značajno je varirao u odnosu na spol, anti-CMV titar, BMI, serumske razine CCL27 i IL-18, i klinički status ispitanika. Manja podskupina CD4+CD8+MR1-tet+TCRVα7.2+ i CD4+CD8- MR1-tet+TCRVα7.2+ MAIT stanica bila je značajno smanjena u perifernoj krvi muških PV ispitanika, u kojih je ujedno zabilježen veći udio Vδ2+γδT stanica s visokom ekspresijom γδTCR receptora, te Vδ1-δ2‐ γδ T stanica s manjom razinom γδTCR ekspresije, dijelom kao odraz razvoja teže kliničke slike. Razine transkripata ZBTB16, IL18R i RUNX3 bile su značajno smanjene u perifernim γδ T stanicama PV pacijenata, dok su MAIT stanice imale različite transkripcijske promjene gena RORC, EOMES i CCR6. Zaključak: Rezultati ovog istraživanja opisuju ranije neprepoznate promjene humanih, cirkulirajućih MAIT i γδ T stanica i otkrivaju njihove promjene u sastavu u ovisnosti o spolu, CMV infekciji, BMI statusu, razinama serumskih citokina te razini upale. Štoviše, zabilježeni su i promijenjeni, divergentni transkripcijski profili sortiranih MAIT i γδ T stanica te povezani s razvojem psorijaze. |
| Abstract (english) | Objectives: The study aimed to determine the phenotype and functional status of mucosa-associated invariant T (MAIT) lymphocytes and gamma delta (γδ) T cells in psoriasis, by studying their peripheral blood proportions, together with IL-17, IL-23, IL-18, CCL20 and CCL27 serum levels, gene expression profiles of chemokine/selectin receptors (IL18R, CCR6, CCR10, SELPLG) and transcription factors likely controlling innate (ZBTB16), cytotoxic/type 1 (RUNX3, EOMES, TBX21), and type 17 functions (RORC) in sorted MAIT and γδ T cells of healthy and affected examinees.. Study design: a case-control study. Participants: the study included 21 adult patients with psoriasis vulgaris (PV) and 22 age- and sex-matched, unrelated healthy controls. Materials and methods: Demographic, clinical – anthropometric, biochemical and data of infective load were collected along with blood and serum samples. Disease severity was determined according to PASI and DLQI questionnaire. Immunophenotyping of MAIT and γδ T cells was performed by multiparametric flow cytometry, including MAIT specific labeling with the 5-OP-RU loaded MR1-tetramer. Luminex technology was used for cytokine and chemokine serum level measurements, and targeted qRT- PCR was performed to determine transcription profile of flow-sorted γδ and MAIT cells. The results of immunophenotyping and genotyping methods were analyzed in relation to the biochemical, serological, demographic and clinical characteristics of the subjects Results: High interpersonal differences in γδ T and MAIT cell composition were observed in relation to sex, anti-CMV titer, BMI, CCL27, and IL-18 serum content and disease severity. A minor subset of canonical CD4+CD8+MR1-tet+TCRVα7.2+ and CD4+CD8-MR1-tet+TCRVα7.2+ T cells was the most significantly underrepresented community in male PV individuals, whereas Vδ2+ γδ T cells expressing high γδTCR and Vδ1-δ2- γδ T cells expressing intermediate γδTCR levels were selectively enriched in affected males, partly reflecting disease severity. The transcript levels of ZBTB16, IL18R, and RUNX3 were significantly reduced in peripheral γδ T cell compartments of PV patients, whereas MAIT cells exhibited differential transcriptional changes of RORC, EOMES, and CCR6. Conclusion: The result of this study demonstrates previously unrecognized skewing of human circulating MAIT and γδ T cell subsets, and reveals their compositional changes in relation to sex, CMV exposure, BMI, serum cytokine content, and inflammatory burden. Moreover, a divergent transcriptional profile of flow-sorted MAIT and γδ cells was described and related to early-stage psoriasis. |
