{"id":1268,"date":"2017-09-28T07:22:10","date_gmt":"2017-09-28T05:22:10","guid":{"rendered":"http:\/\/www.newslab.sk\/2017\/09\/28\/vyuzitie-nepriamej-imunofluorescencie-pri-detekcii-autoprotilatok-asociovanych-so-systemovou-sklerozou\/"},"modified":"2017-10-04T14:23:01","modified_gmt":"2017-10-04T12:23:01","slug":"use-of-indirect-immunofluorescence-in-the-detection-of-autoantibodies-associated-with-systemic-sclerosis","status":"publish","type":"post","link":"https:\/\/www.newslab.sk\/en\/use-of-indirect-immunofluorescence-in-the-detection-of-autoantibodies-associated-with-systemic-sclerosis\/","title":{"rendered":"Use of indirect immunofluorescence in the detection of autoantibodies associated with systemic sclerosis"},"content":{"rendered":"<pre><strong><span style=\"color: #ff0000;\">*All tables, charts, graphs and pictures that are featured in this article can be found in the .pdf \r\nattachment at the end of the paper.<\/span> \r\n<\/strong><\/pre>\n<p>Syst\u00e9mov\u00e1 skler\u00f3za (SSc) \u00a0je zriedkav\u00e9, chronick\u00e9 autoimunitn\u00e9 ochorenie spojivov\u00e9ho tkaniva \u00a0s nezn\u00e1mou pr\u00ed\u010dinou \u00a0vzniku. Charakterizuje ho v\u00fdrazn\u00e1 fibr\u00f3za \u00a0ko\u017ee, \u00a0degenerat\u00edvne zmeny \u00a0a vaskul\u00e1rne abnormality, ktor\u00e9 \u00a0postihuj\u00fa ko\u017eu, k\u013aby a vn\u00fatorn\u00e9 org\u00e1ny \u00a0(najm\u00e4 pa\u017eer\u00e1k, gastrointestin\u00e1lny trakt, p\u013e\u00faca, srdce a obli\u010dky)(1). Na celom svete trp\u00ed t\u00fdmto ochoren\u00edm pribli\u017ene 2 \u2013 50 zo 100\u2009000 \u013eud\u00ed, pri\u010dom \u00a0postihuje 3-4x \u010dastej\u0161ie \u017eeny ako \u00a0mu\u017eov (obvykle medzi \u00a044. a\u017e\u00a0<span style=\"font-size: inherit;\">rokom \u017eivota).\u00a0<\/span>Vzh\u013eadom \u00a0na \u00a0to, \u00a0\u017ee \u00a0SSc \u00a0sa prejavuje r\u00f4znymi \u00a0formami a na r\u00f4znych \u00a0\u010dastiach tela, pri\u010dom \u00a0sa dokonca m\u00f4\u017ee \u00a0dosta\u0165 aj do pokojovej f\u00e1zy, klinick\u00e1 diagn\u00f3za je pomerne n\u00e1ro\u010dn\u00e1(2).<\/p>\n<p>&nbsp;<\/p>\n<p><strong>F<\/strong><strong>ormy SSc:<\/strong><\/p>\n<ol>\n<li><strong> Dif\u00fazna sklerodermia (dcSSc) \u2013 <\/strong>zhrubnutie ko\u017ee na tv\u00e1ri, trupe a na horn\u00fdch i doln\u00fdch kon\u010datin\u00e1ch.<\/li>\n<li><strong>Limitovan\u00e1 sklerodermia (lcSSc) \u2013 <\/strong>zhrubnutie ko\u017ee na obmedzen\u00fdch miestach dist\u00e1lne od \u00a0lak\u0165ov a kolien, ale postihuje tie\u017e tv\u00e1r a Synonymom je CREST syndr\u00f3m (C \u2013 podko\u017en\u00e1 kalcifik\u00e1cia, \u00a0R \u2013 Raynaudov fenom\u00e9n, E \u2013 porucha motility pa\u017eer\u00e1ka, S \u2013 sklerodakt\u00fdlia, T \u2013 teleangiekt\u00e1zia).<\/li>\n<li><strong> Bez ko\u017enej sklerodermie \u2013 <\/strong>bez zjavn\u00fdch \u00a0klinick\u00fdch ko\u017en\u00fdch prejavov (s v\u00fdnimkou tv\u00e1re), ale s charakteristick\u00fdmi zmenami na vn\u00fatorn\u00fdch org\u00e1noch, cievnymi prejavmi a so s\u00e9rologick\u00fdmi n\u00e1lezmi.<\/li>\n<li><strong> Prekr\u00fdvaj\u00faci sa syndr\u00f3m \u2013 <\/strong>SSc sp\u013a\u0148aj\u00faca z\u00e1rove\u0148 aj krit\u00e9ri\u00e1 syst\u00e9mov\u00e9ho lupus erythematosus (SLE), reumatoidnej artrit\u00eddy (RA) alebo polymyozit\u00eddy (PM).<\/li>\n<li><strong> Nediferencovan\u00e9 ochorenie spojivov\u00e9ho tkaniva\/OVER<\/strong><strong>&#8211; <\/strong><strong>L<\/strong><strong>AP <\/strong><strong>syndr\u00f3m \u2013 <\/strong>Raynaudov fenom\u00e9n s klinick\u00fdmi a\/alebo laborat\u00f3rnymi prejavmi SSc (anticentrom\u00e9rov\u00e9 protil\u00e1tky, zmeny \u00a0mikrovaskulat\u00fary, ischemick\u00e9 defekty, ed\u00e9m a ulcer\u00e1cie prstov), \u00a0ale bez n\u00e1lezu \u00a0zhrubnutia ko\u017ee a org\u00e1nov\u00fdch prejavov syst\u00e9movej sklerodermie(6,7).<\/li>\n<\/ol>\n<p>Pr\u00edtomnos\u0165 autoprotil\u00e1tok je charakteristickou \u010drtou \u00a0SSc, ale aj in\u00fdch autoimunitn\u00fdch ochoren\u00ed. Stanovenie autoprotil\u00e1tok v s\u00e9rach pacientov s SSc je zvl\u00e1\u0161\u0165 d\u00f4le\u017eit\u00e9 \u2013 napom\u00e1ha pri sledovan\u00ed priebehu ochorenia, progn\u00f3ze a diferenci\u00e1lnej diagnostike (najm\u00e4 odl\u00ed\u0161enie od in\u00fdch kolagen\u00f3z \u2013 napr. SLE, PM, Sharpov \u00a0syndr\u00f3m, Sj\u04e7grenov syndr\u00f3m)(2,3,4).<\/p>\n<p>Pribli\u017ene v 95 % SSc pr\u00edpadov \u00a0mo\u017eno n\u00e1js\u0165 v s\u00e9re \u00a0pacientov antinukle\u00e1rne protil\u00e1tky (ANA). ANA s\u00fa detegovan\u00e9 met\u00f3dou \u00a0nepriamej imunofluorescencie (NIF) na kultivovan\u00fdch Hep-2 bunk\u00e1ch v kombin\u00e1cii so \u00a0substr\u00e1tom opi\u010dej \u00a0pe\u010dene. T\u00e1to met\u00f3da sl\u00fa\u017ei ako skr\u00edningov\u00fd \u00a0test na detekciu autoprotil\u00e1tok, preto\u017ee je vysokosenzit\u00edvna a poskytuje z\u00e1kladn\u00e9 inform\u00e1cie o detegovanej \u0161trukt\u00fare. Na Hep-2 bunk\u00e1ch mo\u017eno vo v\u0161eobecnosti pop\u00edsa\u0165 fluorescen\u010dn\u00e9 obrazy, napr.\u00a0 homog\u00e9nne jadro, jemne \u00a0zrnit\u00e9\/zrnit\u00e9\/hrubo zrnit\u00e9 jadro, jadierka a in\u00e9 \u0161trukt\u00fary, ktor\u00fdch pr\u00edtomnos\u0165 je sp\u00f4soben\u00e1 r\u00f4znymi antig\u00e9nmi. \u00a0Na potvrdenie konkr\u00e9tneho antig\u00e9nu je indikovan\u00fd imuno\u0161pecifick\u00fd test (ELISA, imunoblot a pod.). \u00a0Imunoblot je met\u00f3da detekcie, pri ktorej s\u00fa na stripoch fixovan\u00e9 \u00a0v\u0161etky v s\u00fa\u010dasnosti zn\u00e1me autoantig\u00e9ny charakteristick\u00e9 pre SSc, ktor\u00e9 sa rozde\u013euj\u00fa do dvoch skup\u00edn: SSc \u0161pecifick\u00e9 (topoizomer\u00e1za I, centrom\u00e9ra-A, centrom\u00e9ra-B, RNA polymer\u00e1za III-RP11, -RP155) a SSc asociovan\u00e9 (fibrilar\u00edn, NOR90, Th\/To, PM-Scl100, PM-Scl75, Ku, PDGFR, Ro-52). Prevalencia autoprotil\u00e1tok proti SSc antig\u00e9nom je uveden\u00e1 v <strong><em>tabu\u013eke 1<\/em><\/strong>(2,3,4,5).<\/p>\n<p>&nbsp;<\/p>\n<p><strong>A<\/strong><strong>utoprotil\u00e1tky proti centrom\u00e9ram \u00a0(anti-CENP)<\/strong><\/p>\n<p>Anti-CENP rozozn\u00e1vaj\u00fa 4 prote\u00edny, naj\u010dastej\u0161ie CENP-A,-B, -C, -D. Centrom\u00e9rov\u00e9 prote\u00edny umo\u017e\u0148uj\u00fa v\u00e4zbu kinetoch\u00f3rov na centrom\u00e9rov\u00fa oblas\u0165 chromoz\u00f3mov a interakciu s vretienkom \u00a0mitotick\u00e9ho apar\u00e1tu po\u010das mit\u00f3zy(5,8).<\/p>\n<p>Anti-CENP sa \u010dastej\u0161ie vyskytuj\u00fa pri lcSSc alebo pri CREST syndr\u00f3me. Pr\u00edtomnos\u0165 t\u00fdchto \u00a0autoprotil\u00e1tok je asociovan\u00e1 s lep\u0161ou progn\u00f3zou a oneskoren\u00fdm n\u00e1stupom postihnutia vn\u00fatorn\u00fdch org\u00e1nov. Tak\u00edto\u00a0 pacienti maj\u00fa \u00a0limitovan\u00e9 ko\u017en\u00e9 ochorenie bez \u00a0postihnutia srdca \u010di obli\u010diek \u00a0a zriedkavo z\u00e1va\u017en\u00fa interstici\u00e1lnu fibr\u00f3zu p\u013e\u00fac (IF). Napriek tomu u viac ako polovice pacientov, ktor\u00ed zomieraj\u00fa v s\u00favislosti s SSc, je pr\u00ed\u010dinou \u00famrtia pokro\u010dil\u00e1 p\u013e\u00facna hypertenzia (PHT). U pacientov s anti-CENP protil\u00e1tkami sa nevyvinie dcSSc \u00a0a v takom \u00a0pr\u00edpade nemusia by\u0165 monitorovan\u00ed pre pr\u00edpadn\u00fa ren\u00e1lnu \u00a0kr\u00edzu a interstici\u00e1lnu fibr\u00f3zu(2,3,4). \u00a0Anti-CENP protil\u00e1tky \u00a0boli n\u00e1jden\u00e9 aj v s\u00e9rach pacientov s SLE, prim\u00e1rnou bili\u00e1rnou cirh\u00f3zou (PBC), RA a so Sj\u04e7grenov\u00fdm syndr\u00f3mom (SS)(4).<\/p>\n<p><strong>Mikroskopick\u00fd obraz NIF ANA: <\/strong>40 \u2013 60 bodiek v interf\u00e1zovom jadre, bodkovan\u00e1 fluorescencia chromat\u00ednovej platni\u010dky po\u010das mit\u00f3zy <strong><em>(obr\u00e1zok 1)<\/em><\/strong>(8). Vzh\u013eadom na to, \u017ee anti-CENP vykazuje charakteristick\u00fd fluorescen\u010dn\u00fd obraz, nie je potrebn\u00e9 potvrdenie imuno\u0161pecifick\u00fdm testom.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>A<\/strong><strong>utoprotil\u00e1tky proti topoizomer\u00e1ze I (anti-Scl70)<\/strong><\/p>\n<p>Scl70 \u00a0(70kDa)\u00a0 je degrada\u010dn\u00fdm produktom topoizomer\u00e1zy-I (topo I). Topo I je enz\u00fdm lokalizovan\u00fd v nukleoplazme, jadierkach a v oblasti organiz\u00e1tora jadierka. \u00a0Jeho funkciou \u00a0je katalyzova\u0165 odv\u00edjanie \u00a0dsDNA pred \u00a0synt\u00e9zou dc\u00e9rskych vl\u00e1ken DNA po\u010das replik\u00e1cie(8).<\/p>\n<p>Henault \u00a0a kol. odhalili priame patog\u00e9nne \u00fa\u010dinky autoprotil\u00e1tok n\u00e1jden\u00fdch v SSc. Vo svojej \u0161t\u00fadii uviedli, \u017ee autoantig\u00e9n \u00a0topo \u00a0I uvo\u013enen\u00fd\u00a0 z apoptotick\u00fdch endoteli\u00e1lnych buniek sa \u0161pecificky via\u017ee na fibroblasty, kde ho u pacientov s SSc rozpozn\u00e1 protil\u00e1tka anti-Scl70. T\u00e1to v\u00e4zba \u00a0n\u00e1sledne indukuje adh\u00e9ziu a aktiv\u00e1ciu monocytov, ktor\u00e1 vedie k v\u00fdvoju SSc(10).<\/p>\n<p>Protil\u00e1tky anti-Scl70 \u00a0s\u00fa u pacientov s SSc pova\u017eovan\u00e9 za marker \u00a0zlej progn\u00f3zy. Ich pr\u00edtomnos\u0165 je asociovan\u00e1 s dcSSc a \u00a0so skor\u00fdm n\u00e1stupom postihnutia vn\u00fatorn\u00fdch org\u00e1nov. U pacientov s t\u00fdmto \u00a0typom \u00a0autoprotil\u00e1tok je vy\u0161\u0161ia \u00a0pravdepodobnos\u0165 v\u00fdskytu IF v skor\u00fdch \u0161t\u00e1di\u00e1ch, perif\u00e9rnej vaskul\u00e1rnej choroby, postihnutia srdca, ren\u00e1lnej \u00a0kr\u00edzy a malign\u00edt. R\u00f4zne \u0161t\u00fadie opisuj\u00fa \u00a0asoci\u00e1ciu protil\u00e1tok anti-Scl70 \u00a0s vysokou \u00famrtnos\u0165ou, pri\u010dom \u00a0pacienti s IF zomieraj\u00fa na komplik\u00e1cie priemerne do 10 rokov po diagnostikovan\u00ed SSc(4,5). Anti-Scl70 sa zvy\u010dajne nevyskytuje u zdrav\u00fdch \u00a0jedincov, ani u pacientov s Raynaudov\u00fdm syndr\u00f3mom alebo\u00a0 s in\u00fdmi ochoreniami spojivov\u00e9ho tkaniva. \u00a0Pravdepodobnos\u0165 v\u00fdskytu anti-Scl70 \u00a0a anti-CENP s\u00fa\u010dasne je ve\u013emi n\u00edzka a vyskytuje sa len u 0,5 % pacientov(3).<\/p>\n<p><strong>Mikroskopick\u00fd obraz NIF ANA: <\/strong>Jemne zrnit\u00e1 a\u017e homog\u00e9nna \u00a0fluorescencia jadra \u00a0v interf\u00e1ze, pozit\u00edvna \u00a0fluorescencia chromat\u00ednovej platni\u010dky \u00a0po\u010das mit\u00f3zy, \u010dasto s pozit\u00edvnymi jadierkami <strong><em>(obr\u00e1zok \u00a02)<\/em><\/strong>(8). Vzh\u013eadom \u00a0na to, \u017ee fluorescen\u010dn\u00fd obraz \u00a0anti-Scl70 \u00a0nie je v\u017edy jednozna\u010dn\u00fd, je potrebn\u00e9 potvrdenie imuno\u0161pecifick\u00fdm testom.<\/p>\n<p><strong>A<\/strong><strong>utoprotil\u00e1tky proti <\/strong><strong>RNA <\/strong><strong>polyme<\/strong><strong>r<\/strong><strong>\u00e1z<\/strong><strong>e<\/strong> <strong>III <\/strong><strong>(anti-RNAP <\/strong><strong>III<\/strong><strong>) <\/strong>RNA polymer\u00e1za III je enz\u00fdm, ktor\u00fd katalyzuje prepis tRNA, \u010das\u0165 rRNA a niektor\u00fdch mal\u00fdch RNA. Ide o komplex prote\u00ednov pozost\u00e1vaj\u00faci z &gt; 12 podjednotiek, napr. RP11 a RP155, ktor\u00e9 s\u00fa asociovan\u00e9 s SSc(11).<\/p>\n<p>Anti-RNAP III sa vyskytuj\u00fa u pacientov s SSc v 5 \u2013 20 % a s\u00fa asociovan\u00e9 s dcSSc \u00a0a progres\u00edvnym zhrubnut\u00edm ko\u017ee, vysok\u00fdm \u00a0rizikom pre v\u00fdvoj ren\u00e1lnej kr\u00edzy, progres\u00edvnej\u0161\u00edm priebehom ochorenia, malignitami, bez \u00a0pr\u00edtomnosti IF. Pre\u017e\u00edvanie pacientov s anti-RNAP III je v porovnan\u00ed s anti-Scl70 lep\u0161ie, preto\u017ee ren\u00e1lna kr\u00edza je v s\u00fa\u010dasnosti \u013eah\u0161ie lie\u010dite\u013en\u00e1 ako IF. Pr\u00edtomnos\u0165 protil\u00e1tok anti-RNAP III u pacientov s SSc m\u00f4\u017ee \u00a0by\u0165 pova\u017eovan\u00e1 za marker \u00a0rozvoja ren\u00e1lnej \u00a0kr\u00edzy(4,5,9,11).<\/p>\n<p><strong>Mikroskopick\u00fd obraz NIF ANA: <\/strong>Jemne zrnit\u00e1 fluorescencia \u00a0jadra \u00a0v interf\u00e1ze bunky, \u00a0chromozom\u00e1lna oblas\u0165 \u00a0po\u010das mit\u00f3zy je negat\u00edvna <strong><em>(obr\u00e1zok \u00a02)<\/em><\/strong>(5,12). Fluorescen\u010dn\u00fd obraz \u00a0pri anti-RNAP III nie je jednozna\u010dne odl\u00ed\u0161ite\u013en\u00fd od in\u00fdch autoprotil\u00e1tok s podobnou fluorescenciou, preto je potrebn\u00e9 potvrdenie imuno\u0161pecifick\u00fdm testom.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>A<\/strong><strong>utoprotil\u00e1tky proti Th\/To<\/strong><\/p>\n<p>Protil\u00e1tky anti-Th\/To \u00a0rozozn\u00e1vaj\u00fa 40kDa prote\u00edn\u00a0 spolo\u010dn\u00fd pre dva mal\u00e9 \u00a0ribonukleoprote\u00edny obsahuj\u00face \u00a07-2 RNA (RN\u00e1-za MRP \u2013 RNA zlo\u017eka mitochondri\u00e1lnej RNA endoribonukle\u00e1zy) a 8-2 RNA (RNA\u00e1za P \u2013 ribonukle\u00e1za P)(8).<\/p>\n<p>Anti-Th\/To \u00a0protil\u00e1tky \u00a0s\u00fa asociovan\u00e9 preva\u017ene s lcSSc formou, \u00a0hoci \u00a0celkov\u00e1 \u00a0frekvencia u pacientov s SSc \u00a0je iba 2 \u2013 5 %. Pacienti s protil\u00e1tkou anti-Th\/To maj\u00fa \u00a0sklon \u00a0ku krat\u0161iemu trvaniu Raynaudov\u00e9ho syndr\u00f3mu pred\u00a0 n\u00e1stupom in\u00fdch sympt\u00f3mov, ako je napr. opuch\u00a0 r\u00fak. Z\u00e1va\u017en\u00e9 ischemick\u00e9 komplik\u00e1cie s\u00fa zriedkav\u00e9, \u00a0av\u0161ak \u00a0pr\u00edtomnos\u0165 tejto protil\u00e1tky sa sp\u00e1ja s IF a PHT, ktor\u00e9 \u00a0sa \u010dasto vyskytuj\u00fa \u00a0u\u017e v skorom \u0161t\u00e1diu ochorenia. T\u00e1to zv\u00fd\u0161en\u00e1 frekvencia a z\u00e1va\u017enos\u0165 p\u013e\u00facnych komplik\u00e1ci\u00ed \u00a0ved\u00fa k hor\u0161iemu pre\u017e\u00edvaniu \u00a0v porovnan\u00ed s pacientmi s lcSSc bez pr\u00edtomnosti tejto protil\u00e1tky(3,4,5).<\/p>\n<p><strong>Mikroskopick\u00fd obraz NIF ANA: <\/strong>Bodkovan\u00e1 fluorescencia jadierok v interf\u00e1ze bunky, chromozom\u00e1lna oblas\u0165 po\u010das mit\u00f3zy je negat\u00edvna(3). Fluorescen\u010dn\u00fd obraz \u00a0pri anti-Th\/To \u00a0nie je jednozna\u010dne odl\u00ed\u0161ite\u013en\u00fd \u00a0od in\u00fdch autoprotil\u00e1tok s podobnou fluorescenciou, preto \u00a0je potrebn\u00e9 potvrdenie imuno\u0161pecifick\u00fdm testom.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>A<\/strong><strong>utoprotil\u00e1tky proti fibrilar\u00ednu\/U3RNP<\/strong><\/p>\n<p>Hlavn\u00fdm \u00a0autoantig\u00e9nom anti-U3RNP je fibrilar\u00edn, 34kDa prote\u00edn, zlo\u017eka U3-ribonukleoprote\u00ednov\u00e9ho komplexu(3,8). Prevalencia antifibrilar\u00ednu \u00a0je u pacientov s SSc 4 \u2013 10 %(5). Anti-U3RNP protil\u00e1tky s\u00fa v\u0161eobecne pova\u017eovan\u00e9 za protil\u00e1tky \u0161pecifick\u00e9 pre\u00a0 SSc \u00a0a vz\u00e1jomne sa vylu\u010duj\u00fa s protil\u00e1tkami anti-CENP, anti-Scl70 a anti-RNAP, boli v\u0161ak \u00a0op\u00edsan\u00e9 aj u pacientov s SLE, nediferencovan\u00fdm ochoren\u00edm spojivov\u00e9ho tkaniva (UCTD) a Raynaudov\u00fdm syndr\u00f3mom. Niektor\u00e9 klinick\u00e9 pr\u00edznaky s\u00fa ovplyvnen\u00e9 \u00a0etnicitou. Belosi, Afroameri\u010dania a Japonci s anti-U3RNP trpia preva\u017ene dcSSc, perif\u00e9rnou vaskulopatiou, vredmi a gangr\u00e9nou. U belochov a Afroameri\u010danov \u00a0je v porovnan\u00ed s Japoncami op\u00edsan\u00e9 \u010dastej\u0161ie postihnutie org\u00e1nov, IF, PHT \u010di ren\u00e1lna kr\u00edza a s nimi s\u00favisiaca hor\u0161ia progn\u00f3za. Pr\u00edtomnos\u0165 anti-U3RNP u pacientov s lcSSc vylu\u010duje rozvoj PHT(3,13).<\/p>\n<p><strong>Mi<\/strong><strong>k<\/strong><strong>r<\/strong><strong>o<\/strong><strong>s<\/strong><strong>ko<\/strong><strong>pic<\/strong><strong>k<\/strong><strong>\u00fd \u00a0obraz \u00a0NIF ANA: <\/strong>Chuchvalcovit\u00e1 \u00a0fluorescencia jadierok\u00a0 v \u00a0interf\u00e1ze, bunky \u00a0v \u00a0mit\u00f3ze vykazuj\u00fa perichromozom\u00e1lnu fluorescenciu (14). Fluorescen\u010dn\u00fd obraz pri anti-U3RNP nie je jednozna\u010dne odl\u00ed\u0161ite\u013en\u00fd od in\u00fdch autoprotil\u00e1tok s podobnou fluorescenciou, preto \u00a0je potrebn\u00e9 potvrdenie imuno\u0161pecifick\u00fdm testom.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>A<\/strong><strong>utoprotil\u00e1tky proti NOR90<\/strong><\/p>\n<p>Protil\u00e1tky anti-NOR90 s\u00fa namieren\u00e9 proti oblasti organiz\u00e1cie jadierka \u00a0(NOR), kde sa po mit\u00f3ze formuj\u00fa \u00a0jadierka. \u00a0\u0160t\u00fadie preuk\u00e1zali, \u017ee autoantig\u00e9nom pre protil\u00e1tku anti-NOR90 je transkrip\u010dn\u00fd faktor RNA polymer\u00e1zy I via\u017euci sa proti smeru transkripcie (hUBF)(5,8).<\/p>\n<p>Anti-NOR90 nepatria medzi \u00a0SSc \u0161pecifick\u00e9 protil\u00e1tky, av\u0161ak boli detegovan\u00e9 u pacientov s ochoreniami spojivov\u00e9ho tkaniva \u00a0ako Raynaudov syndr\u00f3m, RA, SLE, SS a u pacientov s malignitami. \u010coraz \u010dastej\u0161ie sa spom\u00edna asoci\u00e1cia medzi protil\u00e1tkami anti-NOR90 a lcSSc, miernou \u00a0formou \u00a0postihnutia ko\u017ee a priaznivou \u00a0progn\u00f3zou (3,4).<\/p>\n<p><strong>Mikroskopick\u00fd obraz NIF ANA: <\/strong>Zrnit\u00e1 fluorescencia jadierok po\u010das interf\u00e1zy. \u00a0V chromat\u00ednovej oblasti mitotickej bunky pozorovan\u00e9 bodky, ktor\u00e9 \u00a0s\u00fa \u00a0pova\u017eovan\u00e9 za oblas\u0165 \u00a0organiz\u00e1cie jadierka(8). Fluorescen\u010dn\u00fd obraz \u00a0pri anti-NOR90 nie je jednozna\u010dne odl\u00ed\u0161ite\u013en\u00fd \u00a0od in\u00fdch autoprotil\u00e1tok s podobnou fluorescenciou, preto \u00a0je potrebn\u00e9 potvrdenie imuno\u0161pecifick\u00fdm testom.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>A<\/strong><strong>utoprotil\u00e1tky proti PM-Scl75, \u00a0PM-Scl100<\/strong><\/p>\n<p>Autoantig\u00e9n PM-Scl je makromolekulov\u00fd komplex 11 \u2013 16 polypeptidov, z ktor\u00fdch najd\u00f4le\u017eitej\u0161ie maj\u00fa molekulov\u00fa hmotnos\u0165 75kDa a 100kDa:\u00a0 anti-PM-Scl75, \u00a0anti-PM-Scl100. V bunkovom jadre \u00a0sa z\u00fa\u010dast\u0148uje na \u0161tiepen\u00ed prekurzorovej rRNA a v cytoplazme napom\u00e1ha degradova\u0165 zrel\u00fa mRNA(15).<\/p>\n<p>Zatia\u013e \u010do autoprotil\u00e1tky proti CENP a Scl70 s\u00fa typick\u00e9 pre SSc, o autoprotil\u00e1tkach proti PM-Scl75 a PM-Scl100 sa uva\u017euje \u00a0ako \u00a0o s\u00e9rologick\u00fdch markeroch pre \u00a0overlap \u00a0syndr\u00f3m, pri\u010dom \u00a0PM-Scl75 by mohol \u00a0p\u00f4sobi\u0165 \u00a0ako hlavn\u00fd cie\u013eov\u00fd antig\u00e9n pri dcSSc. Anti-PM-Scl protil\u00e1tky \u00a0sa naj\u010dastej\u0161ie vyskytuj\u00fa u pacientov s prekr\u00fdvaj\u00facim sa syndr\u00f3mom PM a SSc (31 %), vz\u00e1cnej\u0161ie pri samotnej myozit\u00edde (8 \u2013 11 %) alebo SSc (2 %)(15,19). Pacienti s t\u00fdmto \u00a0typom autoprotil\u00e1tok \u010dasto trpia \u00a0subak\u00fatnou myozit\u00eddou, \u00a0Raynaudov\u00fdm fenom\u00e9nom a sklerodermiou ko\u017ee(5). \u00a0D\u00f4kaz t\u00fdchto \u00a0autoprotil\u00e1tok je spojen\u00fd s lep\u0161ou progn\u00f3zou ochorenia(15).<\/p>\n<p><strong>Mik<\/strong><strong>r<\/strong><strong>o<\/strong><strong>s<\/strong><strong>k<\/strong><strong>o<\/strong><strong>pi<\/strong><strong>c<\/strong><strong>k<\/strong><strong>\u00fd obraz NIF ANA: <\/strong>Homog\u00e9nna fluorescencia\u00a0 jadierok \u00a0a s\u00fa\u010dasne slab\u0161ia, jemne \u00a0zrnit\u00e1 \u00a0fluorescencia nukleoplazmy v interf\u00e1ze, negat\u00edvna fluorescencia chromat\u00ednovej platni\u010dky po\u010das mit\u00f3zy(14). \u00a0Fluorescen\u010dn\u00fd obraz \u00a0pri anti-PM-Scl75, -100 nie je jednozna\u010dne odl\u00ed\u0161ite\u013en\u00fd od in\u00fdch autoprotil\u00e1tok s podobnou fluorescenciou, preto \u00a0je potrebn\u00e9 potvrdenie imuno\u0161pecifick\u00fdm testom.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>A<\/strong><strong>utoprotil\u00e1tky proti Ku<\/strong><\/p>\n<p>Anti-Ku protil\u00e1tky boli op\u00edsan\u00e9 ako marker overlap syndr\u00f3mu sklerodermie\/PM, av\u0161ak \u00a0zaznamenan\u00fd bol aj ich v\u00fdskyt pri in\u00fdch autoimunitn\u00fdch ochoreniach. Autoantig\u00e9nom pre anti-Ku je prote\u00edn \u00a0Ku s dvoma \u00a0podjednotkami, via\u017euci DNA, z\u00fa\u010dast\u0148uj\u00faci sa na oprave \u00a0dsDNA. Prevalencia anti-Ku protil\u00e1tok u SSc pacientov je 1,5 \u2013 16 %(3,16,17).<\/p>\n<p>Anti-Ku je prim\u00e1rne detegovan\u00e9 u SSc pacientov s overlap syndr\u00f3mom. V\u00e4\u010d\u0161ina pacientov m\u00e1 typick\u00e9 pr\u00edznaky ako Raynaudov fenom\u00e9n, sklerodermia ko\u017ee a obvykle aj lcSSc(5).<\/p>\n<p>Anti-Ku sa zriedkavo vyskytuje aj u pacientov s SLE bez znakov SSc, pri\u010dom \u00a0\u010fal\u0161ie autoprotil\u00e1tky asociovan\u00e9 s SLE, napr. anti-dsDNA, s\u00fa v\u017edy pozit\u00edvne. Menej \u010dasto sa vyskytuje postihnutie vn\u00fatorn\u00fdch org\u00e1nov, ktor\u00e9 je v\u0161ak mierne, zatia\u013e \u010do artrit\u00edda \u00a0je be\u017en\u00e1. Anti-Ku protil\u00e1tka je taktie\u017e \u00a0asociovan\u00e1 s men\u0161\u00edm po\u010dtom vaskul\u00e1rnych prejavov, ako je napr. teleangiekt\u00e1zia(18).<\/p>\n<p><strong>Mik<\/strong><strong>r<\/strong><strong>oskopick<\/strong><strong>\u00fd obraz NIF ANA: <\/strong>Jemne zrnit\u00e1 \u00a0a\u017e \u00a0\u0161kvrnit\u00e1 fluorescencia jadra v interf\u00e1ze, chromozom\u00e1lna oblas\u0165 po\u010das mit\u00f3zy \u00a0je negat\u00edvna. Anti-Ku mo\u017eno odl\u00ed\u0161i\u0165 od in\u00fdch autoprotil\u00e1tok na\u00a0 z\u00e1klade substr\u00e1tu opi\u010dej \u00a0pe\u010dene, na\u00a0 kto- rom \u00a0vykazuje \u00a0perinukle\u00e1rnu \u00a0a\u017e \u00a0chum\u00e1\u010dikovit\u00fa fluorescenciu(8). Vzh\u013eadom \u00a0na to, \u017ee fluorescen\u010dn\u00fd obraz \u00a0anti-Ku nie je v\u017edy jednozna\u010dn\u00fd, je potrebn\u00e9 potvrdenie imuno\u0161pecifick\u00fdm testom.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>A<\/strong><strong>utoprotil\u00e1tky proti PDGFR (platelet derived growth factor \u2013 rastov\u00fd faktor odvoden\u00fd od trombocytov)<\/strong><\/p>\n<p>Anti-PDGFR patria medzi protil\u00e1tky asociovan\u00e9 s SSc a boli detegovan\u00e9 aj pri in\u00fdch ochoreniach: SLE, SS, RA a IF(4). Jedn\u00fdm z prv\u00fdch prejavov v patogen\u00e9ze SSc je mikrovaskulat\u00f3rne po\u0161kodenie a akumul\u00e1cia buniek exprimuj\u00facich receptor PDGFR. Autoprotil\u00e1tky anti-PDGFR hraj\u00fa \u00falohu v patogen\u00e9ze SSc, preto\u017ee konvertuj\u00fa \u00a0norm\u00e1lne fibroblasty <em>in vitro <\/em>na bunky podobn\u00e9 SSc, ktor\u00e9 s\u00fa schopn\u00e9 indukova\u0165 fibr\u00f3zu <em>in vivo. <\/em>\u010eal\u0161ie \u0161t\u00fadie t\u00fdchto \u00a0protil\u00e1tok m\u00f4\u017eu pom\u00f4c\u0165 \u00a0v prevencii \u00a0z\u00e1va\u017en\u00fdch \u00a0komplik\u00e1ci\u00ed, \u00a0ako s\u00fa\u00a0 ulcer\u00e1cie, PHT a ren\u00e1lna kr\u00edza, ktor\u00e9 s\u00fa zodpovedn\u00e9 za podstatn\u00e9 skr\u00e1tenie \u010dasu pre\u017e\u00edvania a zhor\u0161enie kvality \u017eivota(20).<\/p>\n<p><strong>Mik<\/strong><strong>r<\/strong><strong>oskopick<\/strong><strong>\u00fd obraz NIF ANA: <\/strong>Nie je op\u00edsan\u00fd \u0161pecifick\u00fd fluorescen\u010dn\u00fd obraz, \u00a0preto \u00a0jedin\u00e1 \u00a0mo\u017enos\u0165 d\u00f4kazu anti-PDGFR je imuno\u0161pecifick\u00fd test.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>Z\u00e1ver<\/strong><\/p>\n<p>Autoprotil\u00e1tky \u00a0detegovan\u00e9 pri SSc \u00a0s\u00fa ve\u013emi \u0161pecifick\u00e9 a asociovan\u00e9 s r\u00f4znymi klinick\u00fdmi prejavmi pacienta. Ich pr\u00edtomnos\u0165 by mala by\u0165 zv\u00e1\u017een\u00e1 pri monitorovan\u00ed pacienta, stanoven\u00ed lie\u010dby a \u010fal\u0161ej \u00a0progn\u00f3ze. Objasnenie \u0161pecificity SSc protil\u00e1tok a s nimi s\u00favisiacich odli\u0161n\u00fdch klinick\u00fdch prejavov by mohlo napom\u00f4c\u0165 lep\u0161ie \u00a0pochopi\u0165 \u00a0patogen\u00e9zu SSc.\u00a0 V\u00fdznam autoprotil\u00e1tok pri SSc ochoren\u00ed je st\u00e1le pomerne nejasn\u00fd, aj ke\u010f \u010doraz viac \u0161t\u00fadi\u00ed nasved\u010duje tomu, \u017ee tieto protil\u00e1tky nie s\u00fa len diagnostick\u00fdmi markermi ochorenia, ale taktie\u017e vedia \u00a0predpoveda\u0165 v\u00fdvoj ochorenia \u010di pr\u00edpadn\u00e9 postihnutie org\u00e1nov.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>L<\/strong><strong>I<\/strong><strong>TER<\/strong><strong>A<\/strong><strong>T\u00daRA<\/strong><\/p>\n<ol>\n<li>Rovensk\u00fd J, Va\u0161\u00e1kov\u00e1 \u00a0M. Sine syndr\u00f3m \u00a0pri syst\u00e9movej skler\u00f3ze. Via practica. 2010; 7(6): 262-265.<\/li>\n<li>EUROIMMUN: Protil\u00e1tky proti specifick\u00fdm antig\u00e9num syst\u00e9mov\u00e9 skler\u00f3zy. N\u00e1vod na proveden\u00ed \u00a0testu Systemic Sclerosis (Nucleoli) Profile EUROLINE (IgG).<\/li>\n<li>Hamaguchi Y. Autoantibody\u00a0 profiles in systemic sclerosis: Predictive value for clinical evaluation \u00a0and prognosis. Journal of Dermatology 2010; 37: 42-53.<\/li>\n<\/ol>\n<ol start=\"4\">\n<li>Wielosz E, Dryglewska M, Majdan \u00a0M. Serological \u00a0profile of patients with \u00a0systemic sclerosis. \u00a0Postepy Hig Med \u00a0Dosw \u00a0(online) \u00a02014; \u00a068: 987-991. e-ISSN 1732-2693.<\/li>\n<\/ol>\n<ol start=\"5\">\n<li>Kuwana M. Circulating anti-nuclear antibodies in Systemic sclerosi: utility in diagnosis and disease subsetting. J Nippon Med Sch 2017; 84(2): <span style=\"font-size: inherit;\">56-63.<\/span><\/li>\n<\/ol>\n<ol start=\"6\">\n<li>Be\u010dv\u00e1\u0159 R. Sou\u010dasn\u00fd pohled na diagnostiku a l\u00e9\u010dbu syst\u00e9mov\u00e9 sklerodermie a Sj\u00f6grenova syndromu. Med Pro Praxi 2008; 5(3): 109-112.<\/li>\n<\/ol>\n<ol start=\"7\">\n<li>Shoenfeld Y, Meroni PL. The General Practice Guide to Autoimmune Diseases. Pabst Science Publishers 2012: ISBN 978-3-89967-770-6<\/li>\n<\/ol>\n<ol start=\"8\">\n<li>Bradwell AR, Hughes \u00a0RG. Atlas of HEp-2 patterns. The Binding Site Ltd. 2007: ISBN 0704425955.<\/li>\n<\/ol>\n<ol start=\"10\">\n<li>Steen \u00a0VD. Autoantibodies \u00a0in Systemic Sclerosis. \u00a0Semin \u00a0Arthritis Rheum 2005; 35: 35-42.<\/li>\n<li>Henault J, Robitaille G, Senecal JL, a spol. DNA topoisomerase I binding to fibroblasts induces monocyte adhesion and activation in the presence of antiopoisomerase I autoantibodies from systemic sclerosis patients. \u00a0<em>A<\/em>rthritis Rheum 2006; 54: 963-973.<\/li>\n<li>Kuwana M, Kimura K, Kawakami Y. 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Arthritis Research\u2008&amp;\u2008therapy 2003; 5(2): 80-93.<\/li>\n<\/ol>\n<ol start=\"14\">\n<li>https:\/<a href=\"http:\/\/www.euroimmun.com\/documents\/Indications\/Autoimmuni-\">\/w<\/a>w<a href=\"http:\/\/www.euroimmun.com\/documents\/Indications\/Autoimmuni-\">w.euroimmun.com\/documents\/Indications\/Autoimmuni<\/a>ty\/Rheumatology\/ANA\/FA_1510_I_UK_B.pdf<\/li>\n<\/ol>\n<ol start=\"15\">\n<li>Ple\u0161tilov\u00e1 L, \u00a0Vencovsk\u00fd \u00a0J. \u00a0Anti-PM-Scl \u00a0protil\u00e1tky \u00a0u myositid. \u00a0\u010ces Revmatol 2009; 17(4): 200-204.<\/li>\n<\/ol>\n<ol start=\"16\">\n<li>Hoa S, Hudson M, Troyanov Y, a spol. \u00a0Single-specificity \u00a0anti-Ku antibodies \u00a0in an international cohort \u00a0of 2140 \u00a0systemic sclerosis subjects: clinical associations. Medicine 2016; 95: 35.<\/li>\n<li>Mimori T, Akizuki M, Yamagata H, a spol. Characterization of a high molecular weight acidic nuclear protein recognized by autoantibodies in sera from patients with polymyositis-scleroderma overlap. <em>J Clin Investig.\u00a0<\/em>1981; 68: 611-620.<\/li>\n<\/ol>\n<ol start=\"18\">\n<li>Rozman B, Cucnik S, Sodin-Semrl S, a spol. Prevalence and clinical associations of anti-Ku antibodies in patients with systemic sclerosis: a European EUSTAR-initiated multi-centre case-control study. Ann Rheum Dis 2008; 67: 1282-1286.<\/li>\n<\/ol>\n<ol start=\"19\">\n<li><strong>\u00a0<\/strong><a href=\"http:\/\/arthritis-research.biomedcentral.com\/articles\/10.1186\/ar2614\">http:\/\/arthritis-researbiomedcentral.com\/articles\/10.1186\/ar2614<\/a><\/li>\n<li>Svegliati S, Amico D, Spadoni T, a spol. Agonistic Anti-PDGFR receptor autoantibodies from patients with systemic sclerosis impact \u00a0human pulmonary \u00a0artery smooth muscle cells function \u00a0<em>in vitro. <\/em>Frontiers \u00a0in Im- munology 2017; 8: 75.<\/li>\n<li>Arch\u00edv Imunol\u00f3gia, Medirex.<\/li>\n<li>https:\/<a href=\"http:\/\/www.researchgate.net\/figure\/45281682_fig7_Fig-8-Fine-\">\/ww<\/a>w<a href=\"http:\/\/www.researchgate.net\/figure\/45281682_fig7_Fig-8-Fine-\">.researchgate.net\/figure\/45281682_fig7_Fig-8-Fine- <\/a>grainy-nucleoplasmic-staining-Scl-70-like<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"<p>*All tables, charts, graphs and pictures that are featured in this article can be found in the .pdf attachment at the end of the paper. Syst\u00e9mov\u00e1 skler\u00f3za (SSc) \u00a0je zriedkav\u00e9, chronick\u00e9 autoimunitn\u00e9 ochorenie spojivov\u00e9ho tkaniva \u00a0s nezn\u00e1mou pr\u00ed\u010dinou \u00a0vzniku. Charakterizuje ho v\u00fdrazn\u00e1 fibr\u00f3za \u00a0ko\u017ee, \u00a0degenerat\u00edvne zmeny \u00a0a vaskul\u00e1rne abnormality, ktor\u00e9 \u00a0postihuj\u00fa ko\u017eu, k\u013aby a vn\u00fatorn\u00e9<\/p>\n","protected":false},"author":7,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_mi_skip_tracking":false,"footnotes":""},"categories":[291,287],"tags":[537,535,538],"class_list":["post-1268","post","type-post","status-publish","format-standard","hentry","category-imunology","category-uncategorized","tag-autoantibodies","tag-autoantigen-en","tag-systemic-sclerosis","typ_clanku-review-article"],"acf":{"abstrakt":"<p>Systemic sclerosis is an autoimmune disease characterised by the presence of antinuclear antibodies against various extractable antigens. In patient\u2019s sera, \u00a0we can detect autoantibodies specific for systemic sclerosis (au- toantibodies against topoisomerase I, centromere-A, centromere-B, RNA polymerase III -RP11, RP155) \u00a0and au- toantibodies associated with systemic sclerosis (autoantibodies against fibrillarin, NOR90, Th\/To, \u00a0PM-Scl100, PM-Scl75, \u00a0Ku, PDGFR, Ro-52). Individual types of autoantibodies may be associated with various clinical mani- festations of the patient, which may help in the diagnosis and management of the disease.<\/p>\n<p><strong>K<\/strong><strong>e<\/strong><strong>ywords: <\/strong>autoantibodies, autoantigen, systemic sclerosis<\/p>\n","casopis":[{"ID":1223,"post_author":"7","post_date":"2017-09-26 14:15:30","post_date_gmt":"2017-09-26 12:15:30","post_content":"<ul>\r\n \t<li>Fertility disorders: immunological causes and possible curative impact<\/li>\r\n \t<li>The first results of galactose-deficient IgA1 measurement in diagnosis and monitoring of patients with IgA nephropathy<\/li>\r\n \t<li>Application of autovaccines in the treatment of chronic and recurrent colpitises<\/li>\r\n \t<li>Infections of the hip endoprostheses<\/li>\r\n \t<li>Non-invasive markers of liver fibrosis<\/li>\r\n<\/ul>","post_title":"Newslab","post_excerpt":"","post_status":"publish","comment_status":"closed","ping_status":"closed","post_password":"","post_name":"newslab-2017-2","to_ping":"","pinged":"","post_modified":"2017-09-26 14:19:35","post_modified_gmt":"2017-09-26 12:19:35","post_content_filtered":"","post_parent":0,"guid":"http:\/\/www.newslab.sk\/casopis\/newslab-2017-2\/","menu_order":0,"post_type":"casopis","post_mime_type":"","comment_count":"0","filter":"raw"}],"strana":"94","upload_clanok":{"ID":1270,"id":1270,"title":"NEWSLAB 2-2017_Durankov\u00e1 PP","filename":"NEWSLAB-2-2017_Durankov\u00e1-PP.pdf","filesize":851119,"url":"https:\/\/www.newslab.sk\/wp-content\/uploads\/2017\/09\/NEWSLAB-2-2017_Durankov\u00e1-PP.pdf","link":"https:\/\/www.newslab.sk\/en\/use-of-indirect-immunofluorescence-in-the-detection-of-autoantibodies-associated-with-systemic-sclerosis\/newslab-2-2017_durankova-pp\/","alt":"","author":"7","description":"","caption":"","name":"newslab-2-2017_durankova-pp","status":"inherit","uploaded_to":1268,"date":"2017-09-28 06:11:35","modified":"2017-09-28 06:11:35","menu_order":0,"mime_type":"application\/pdf","type":"application","subtype":"pdf","icon":"https:\/\/www.newslab.sk\/wp-includes\/images\/media\/document.png"}},"_links":{"self":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1268","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/users\/7"}],"replies":[{"embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/comments?post=1268"}],"version-history":[{"count":0,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1268\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/media?parent=1268"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/categories?post=1268"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/tags?post=1268"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}