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A Pikkelysömör Klinikai, Biokémiai, Immunológiai És Genetikai Jellegzetességeinek Vizsgálata

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A Pikkelysömör Klinikai, Biokémiai, Immunológiai És Genetikai Jellegzetességeinek Vizsgálata dc_1289_16 MTA DOKTORI ÉRTEKEZÉS A PIKKELYSÖMÖR KLINIKAI, BIOKÉMIAI, IMMUNOLÓGIAI ÉS GENETIKAI JELLEGZETESSÉGEINEK VIZSGÁLATA Dr. Gyulai Rolland Pécsi Tudományegyetem, ÁOK Bőr-, Nemikórtani és Onkodermatológiai Klinika PÉCS 2016 Powered by TCPDF (www.tcpdf.org) dc_1289_16 Mottó: A csodálkozás a tudás magja Francis Bacon 2 Powered by TCPDF (www.tcpdf.org) dc_1289_16 TARTALOMJEGYZÉK RÖVIDÍTÉSEK JEGYZÉKE ..................................................................................................................... 5 1. ELŐSZÓ ......................................................................................................................................... 7 2. IRODALMI ÁTTEKINTÉS ............................................................................................................... 8 2.1. Bevezetés, a pikkelysömör jelentősége, a betegség által okozott terhek ............................... 8 2.2. A pikkelysömör és a pikkelysömör kezelésének történeti áttekintése ..................................... 9 2.3. A pikkelysömör klinikai jellemzői, tünetei ................................................................................ 9 2.3.1. A pikkelysömör bőrtünetei, klinikai megjelenési formái ............................................................... 10 2.3.2. A pikkelysömör körömtünetei ...................................................................................................... 12 2.3.3. A pikkelysömörhöz társuló komorbiditások .................................................................................. 12 2.3.4. A pikkelysömör provokáló faktorai ............................................................................................... 15 2.3.5. A pikkelysömör súlyosságának meghatározása .......................................................................... 16 2.4. A pikkelysömör genetikai háttere ........................................................................................... 17 2.4.1. Familiáris halmozódás és öröklésmenet...................................................................................... 17 2.4.2. Psoriasisra hajlamosító gének ..................................................................................................... 17 2.4.3. Arthritis psoriatica kialakulására hajlamosító gének .................................................................... 19 2.5. A pikkelysömör immunpatogenezise ..................................................................................... 19 2.5.1. A pikkelysömörös bőr immunhisztológiai jellegzetességei .......................................................... 19 2.5.2. A veleszületett immunrendszer szerepe a tünetek iniciációjában ................................................ 20 2.5.3. A T-sejtek szerepe a plakk fenntartásában ................................................................................. 21 2.5.4. A hám barrier és a bőr funkcionális károsodásának jelentősége ................................................. 21 2.5.5. A pikkelysömör patogenezisének kompex modellje .................................................................... 22 2.6. A pikkelysömör kezelése ....................................................................................................... 22 2.6.1. A psoriasis kezelésében alkalmazott lokális készítmények ......................................................... 22 2.6.2. A psoriasis kezelésében alkalmazott fizikai eljárások – fényterápia ............................................ 23 2.6.3. A psoriasis kezelésében alkalmazott szisztémás készítmények ................................................. 24 3. CÉLKITŰZÉSEK ........................................................................................................................... 28 4. MÓDSZEREK ............................................................................................................................... 29 4.1. Betegek, kontrollok, kísérleti állatok ...................................................................................... 29 4.1.1. Humán vizsgálatok ...................................................................................................................... 29 4.1.2. TRPA1 KO és kontroll egerek ..................................................................................................... 32 4.2. Klinikai vizsgálatok és kérdőíves felmérések ........................................................................ 32 4.2.1. Kelet Európai psoriasisos betegeken végzett felmérés ............................................................... 32 4.2.2. A dohányzás szerepe a periodontitis kialakulásában psoriasisban ............................................. 32 4.2.3. A methotrexát kezelés klinikai gyakorlatának felmérése psoriasisban ........................................ 33 4.3. Sejtek szeparálása, sejtkultúra, proliferációs vizsgálatok ..................................................... 33 4.3.1. T-sejtek, antigén prezentáló sejtek, bőr sejtek szeparálása ........................................................ 33 4.3.2. T-sejt proliferációs kísérletek ....................................................................................................... 34 4.3.3. Sejtosztódási vizsgálatok ............................................................................................................ 34 4.4. Molekuláris biológiai vizsgálatok ........................................................................................... 35 4.4.1. RNS, DNS izolálás ...................................................................................................................... 35 4.4.2. PCR, RT-PCR ............................................................................................................................. 35 4.4.3. Polimorfizmus vizsgálatok ........................................................................................................... 35 4.5. Áramlási citometriás vizsgálatok ........................................................................................... 36 3 Powered by TCPDF (www.tcpdf.org) dc_1289_16 4.6. Gyengített teljes reflexiós Fourier transzformációs infravörös (ATR-FTIR) spektroszkópia . 37 4.7. ELISA vizsgálatok .................................................................................................................. 37 4.7.1. Citrullinált peptidek koncentrációjának meghatározása ............................................................... 37 4.7.2. II. típusú szolubilis IL-1 receptor (sIL-1R2) koncentráció meghatározása ................................... 37 4.7.3. TNF, TNF-inhibitor, anti-TNF-inhibitor antitest koncentráció meghatározása .............................. 37 4.8. Imiquimod kiváltotta psoriasis egér modell ............................................................................ 38 4.8.1. Psoriasiform bőrgyulladás kiváltása imiquimod segítségével ...................................................... 38 4.8.2. Hátbőr duzzadás (ödéma/infiltráció) mértékének meghatározása ............................................... 39 4.8.3. A bőrszövet perfúziójának mérése .............................................................................................. 39 4.8.4. Lépmegnagyobbodás vizsgálata ................................................................................................. 39 4.8.5. Szöveti citokin mRNS expressziós vizsgálatok ........................................................................... 39 4.8.6. Szöveti citokin fehérjék meghatározása ...................................................................................... 40 4.8.7. Szövettani vizsgálatok ................................................................................................................. 40 4.9. Statisztikai módszerek ........................................................................................................... 41 5. EREDMÉNYEK ÉS MEGBESZÉLÉS ........................................................................................... 42 5.1. A psoriasis genetikai hátterének vizsgálata .......................................................................... 42 5.1.1. A TNFSF15 gén polimorfizmusainak vizsgálata psoriasisban ..................................................... 42 5.1.2. Az ERAP1/HLA-C gének polimorfizmusainak vizsgálata psoriasisban ....................................... 45 5.2. A pikkelysömör immunpatogenezisének felderítésére irányuló vizsgálatok ......................... 51 5.2.1. A tünetmentes psoriasisos bőr fehérje szerkezetének vizsgálata ............................................... 51 5.2.2. A regulátor T-sejtek működésének vizsgálata psoriasisban ........................................................ 57 5.2.3. A humán Treg sejtek azonosítására és szeparálására alkalmas sejtfelszíni marker azonosítása72 5.2.4. Az IL-1R expresszió jelentőségének vizsgálata psoriasisos T-sejtek működésében .................. 75 5.2.5. A bőr TRPA1+ nociceptív neuronjainak szerepe az imiquimod-indukált psoriasiform bőrgyulladásban ........................................................................................................................................... 84 5.3. A psoriasis klinikai jellemzőinek vizsgálata ........................................................................... 94 5.3.1. A betegség súlyossága, az életminőség és a kezelési szokások felmérése kelet-közép-európai psoriasisos betegeken ................................................................................................................................
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