{"id":2247,"date":"2021-09-20T07:59:25","date_gmt":"2021-09-20T05:59:25","guid":{"rendered":"https:\/\/www.newslab.sk\/?p=2247"},"modified":"2021-09-20T08:21:50","modified_gmt":"2021-09-20T06:21:50","slug":"contribution-of-immunohistochemistry-to-histomorphological-diagnostics-of-covid-19-pneumonia","status":"publish","type":"post","link":"https:\/\/www.newslab.sk\/en\/contribution-of-immunohistochemistry-to-histomorphological-diagnostics-of-covid-19-pneumonia\/","title":{"rendered":"Contribution of immunohistochemistry to histomorphological diagnostics of COVID-19 pneumonia"},"content":{"rendered":"

*A rare case of autochthonous human dirofilariasis with the manifestation of pseudotumor of the epididymis caused by helminth Dirofilaria repens<\/strong><\/span><\/p>\n

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\u00davod<\/strong><\/p>\n

Ochorenie COVID-19 sa za\u010dalo \u0161\u00edri\u0165 koncom roka 2019 z \u010d\u00ednskeho mesta Wu-chan v provincii Chu-pej. Ku koncu marca 2021 bolo celosvetovo zaznamenan\u00fdch viac ako 132 mili\u00f3nov potvrden\u00fdch pr\u00edpadov n\u00e1kazy, pri\u010dom na ochorenie COVID-19 zomreli viac ako 3 mili\u00f3ny \u013eud\u00ed(1). Prejavy ochorenia var\u00edruj\u00fa od asymptomatick\u00fdch pr\u00edpadov cez pacientov s typick\u00fdmi chr\u00edpkov\u00fdmi pr\u00edznakmi a\u017e po v\u00e1\u017ene pr\u00edpady z\u00e1palu p\u013e\u00fac, neraz s tragick\u00fdm koncom. Priebeh ochorenia nemo\u017eno spo\u013eahlivo predv\u00edda\u0165, ale medzi fat\u00e1lnymi pr\u00edpadmi dominuj\u00fa pacienti s vy\u0161\u0161\u00edm vekom, mu\u017esk\u00e9ho pohlavia a s komorbiditami ako arteri\u00e1lna hypertenzia, diabetes mellitus, obezita, chronick\u00e9 ochorenie p\u013e\u00fac, srdca a in\u00fdch org\u00e1nov, n\u00e1dorov\u00e9 ochorenie a klinicky v\u00fdznamn\u00e1 imunodeficiencia(2).<\/p>\n

Pre spr\u00e1vny terapeutick\u00fd pr\u00edstup pri v\u00e1\u017enych pr\u00edpadoch je nevyhnutn\u00e9 pozna\u0165 patogen\u00e9zu ochorenia. Klinick\u00e9 \u00fadaje dop\u013a\u0148a, upres\u0148uje aj objas\u0148uje patologickoanatomick\u00fd n\u00e1lez. Jeho nenahradite\u013en\u00e1 \u00faloha sa u\u017e potvrdila aj pri in\u00fdch infek\u010dn\u00fdch ochoreniach v minulosti(9). V pr\u00edpade COVID-19 najz\u00e1va\u017enej\u0161ie d\u00f4sledky vypl\u00fdvaj\u00fa z po\u0161kodenia p\u013e\u00facneho parench\u00fdmu s rozvojom interstici\u00e1lneho, v\u00edrusov\u00e9ho z\u00e1palu p\u013e\u00fac. Skor\u00e9 i neskor\u00e9 zmeny maj\u00fa typick\u00fd makroskopick\u00fd aj mikroskopick\u00fd obraz.<\/p>\n

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Patologickoanatomick\u00fd n\u00e1lez pri COVID-19 pneum\u00f3nii<\/h2>\n

Makroskopick\u00fd n\u00e1lez<\/strong><\/h3>\n

Makroskopicky s\u00fa p\u013e\u00faca vo v\u0161eobecnosti \u0165a\u017ek\u00e9 a edemat\u00f3zne, s n\u00e1lezom dif\u00faznej alebo lo\u017eiskovej konsolid\u00e1cie(3,10). V pr\u00edpade COVID-19 je \u010dast\u00fd n\u00e1lez lo\u017e\u00edsk krv\u00e1cania a infarktov parench\u00fdmu, nezriedka s n\u00e1lezom tromb\u00f3zy pr\u00edvodn\u00fdch ciev(11). To sa prejav\u00ed tmavofialov\u00fdm sfarben\u00edm na pozad\u00ed sivo\u010derven\u00e9ho elastick\u00e9ho z\u00e1palovo postihnut\u00e9ho tkaniva (obr\u00e1zok 1)<\/strong>. N\u00e1lez pleur\u00e1lneho v\u00fdpotku nie je pre CO- VID-19 typick\u00fd(11).<\/p>\n

Tracheobronchit\u00edda sa vyskytuje u 88 % pacientov s let\u00e1lnym priebehom COVID-19 nez\u00e1visle od intub\u00e1cie a superinfekcie. Makroskopick\u00fdm korel\u00e1tom s\u00fa drobn\u00e9 sivobelav\u00e9 aft\u00f3zne ulcer\u00e1cie s priemerom 2-3 mm, v niektor\u00fdch pr\u00edpadoch iba fok\u00e1lne(10).<\/p>\n

Nezvy\u010dajne \u010dast\u00fdm n\u00e1lezom je mas\u00edvna p\u013e\u00facna emboliz\u00e1cia z hlbok\u00fdch \u017e\u00edl doln\u00fdch kon\u010dat\u00edn, ale aj \u010derstv\u00e1 \u017eilov\u00e1 tromb\u00f3za v tejto oblasti bez emboliz\u00e1cie do p\u013e\u00fac(3,5).<\/p>\n

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Mikroskopick\u00fd n\u00e1lez<\/h3>\n

Mikroskopicky sa v\u00edrusov\u00e9 pneum\u00f3nie prejavuj\u00fa r\u00f4znymi morfologick\u00fdmi zmenami, obvykle ako dif\u00fazne alveol\u00e1rne po\u0161kodenie (DAD, diffuse alveolar damage). Ide o naj\u010dastej\u0161\u00ed histologick\u00fd korel\u00e1t klinickej diagn\u00f3zy syndr\u00f3mu ak\u00fatnej respira\u010dnej tiesne (ARDS, acute respiratory distress syndrome). Tak ako klinicky, aj mikroskopicky ide o ne\u0161pecifick\u00fd obraz, ktor\u00fd m\u00f4\u017ee by\u0165 sp\u00f4soben\u00fd napr\u00edklad aj toxick\u00fdmi vplyvmi, bakt\u00e9riami alebo hubami(6,11). Na v\u00edrusov\u00fd p\u00f4vod DAD poukazuje n\u00e1lez cytopatick\u00e9ho efektu v\u00edrusov, a to najm\u00e4 intranukle\u00e1rnych, intracytoplazmov\u00fdch inkl\u00fazi\u00ed a mnohojadrov\u00fdch buniek. Tieto s\u00fa pri niektor\u00fdch v\u00edrusov\u00fdch pneum\u00f3ni\u00e1ch vyvolan\u00fdch p\u00f4vodcami ako cytomegalov\u00edrus alebo respira\u010dn\u00fd syncyci\u00e1lny v\u00edrus ve\u013emi v\u00fdrazn\u00e9, ale pri mnoh\u00fdch in\u00fdch, ako napr\u00edklad chr\u00edpka alebo SARS, ich nenach\u00e1dzame(6). In\u00fdmi typick\u00fdmi n\u00e1lezmi s\u00fa ak\u00fatna bronchit\u00edda, organizuj\u00faca sa pneum\u00f3nia a dif\u00fazna interstici\u00e1lna pneum\u00f3nia(6).<\/p>\n

V pr\u00edpade COVID-19 s\u00fa typick\u00fdm n\u00e1lezom najm\u00e4 znaky exsudat\u00edvnej a proliferat\u00edvnej f\u00e1zy DAD v r\u00f4znom \u0161t\u00e1diu s po\u0161koden\u00edm bronchi\u00e1lneho epitelu, \u010dasto s progresiou do fibr\u00f3zy(3). Rozvoju DAD v skor\u00fdch \u0161t\u00e1di\u00e1ch predch\u00e1dza n\u00e1lez ak\u00fatnej alveol\u00e1rnej kapilarit\u00eddy sprev\u00e1dzanej zmie\u0161anou z\u00e1palovou infiltr\u00e1ciou alveol\u00e1rnych priehradiek lymfocytmi a neutrofilmi (obr\u00e1zky 2, 3C)<\/strong>(11), a to aj bez n\u00e1padnej\u0161ieho po\u0161kodenia mimop\u013e\u00facnych ciev(10). Histomorfologick\u00e1 diagnostika COVID-19 pneum\u00f3nie v inici\u00e1lnom \u0161t\u00e1diu v\u00fdvoja je \u0165a\u017ek\u00e1 a v pr\u00edpade, \u017ee pitva bola vykonan\u00e1 do 30 hod\u00edn po smrti, imunohistochemick\u00e1 detekcia SARS-CoV-2 v\u00edrusovej infekcie m\u00f4\u017ee by\u0165 k\u013e\u00fa\u010dov\u00e1 pre stanovenie spr\u00e1vneho diagnostick\u00e9ho z\u00e1veru (obr\u00e1zok 2)<\/strong>.<\/p>\n

N\u00e1lezy typick\u00e9 pre exsudat\u00edvnu f\u00e1zu DAD, op\u00edsan\u00e9 pri ochoren\u00ed COVID-19, zah\u0155\u0148aj\u00fa kongesciu mal\u00fdch ciev, zhrubnutie alveol\u00e1rnych sept, intraalveol\u00e1rny ed\u00e9m(3,8,10), pri\u010dom osobitne op\u00edsan\u00fd je prote\u00ednov\u00fd exsud\u00e1t v alveol\u00e1rnych priestoroch s ojedinel\u00fdmi ve\u013ek\u00fdmi prote\u00ednov\u00fdmi globulami a intraalveol\u00e1rny fibr\u00edn(5). Typick\u00e9 je formovanie hyal\u00ednov\u00fdch membr\u00e1n, perivaskul\u00e1rna z\u00e1palov\u00e1 infiltr\u00e1cia lymfocytmi a plazmatick\u00fdmi bunkami a lo\u017eisk\u00e1 intraalveol\u00e1rneho krv\u00e1cania(3,8,10) (obr\u00e1zok 3)<\/strong>. Charakteristick\u00fdm n\u00e1lezom je aj deskvam\u00e1cia epitelu a aktivovan\u00e9 pneumocyty II. typu s v\u00fdrazne<\/p>\n

zv\u00e4\u010d\u0161en\u00fdm jadrom a prominentn\u00fdm jadierkom a ojedinel\u00e9 mnohojadrov\u00e9 syncyci\u00e1lne bunky vznikaj\u00face ich f\u00faziou(5,8,10) (obr\u00e1zky 3A, 4)<\/strong>. Niektor\u00ed autori op\u00edsali n\u00e1lez intracytoplazmov\u00fdch inkl\u00fazi\u00ed, ich v\u00edrusov\u00fd p\u00f4vod v\u0161ak nebol jednozna\u010dne potvrden\u00fd(8,10). Nukle\u00e1rne inkl\u00fazie op\u00edsan\u00e9 neboli(10), zriedkavo v\u0161ak mo\u017eno n\u00e1js\u0165 ve\u013ek\u00e9 epitelov\u00e9 bunky s n\u00e1padne ve\u013ek\u00fdm jadierkom, \u010do by mohlo by\u0165 v\u00fdsledkom v\u00edrusov\u00e9ho cytopatick\u00e9ho \u00fa\u010dinku vzh\u013eadom na imunohistochemicky potvrden\u00fa pr\u00edtomnos\u0165 SARS-CoV-2 v\u00edrusovej infekcie v tak\u00fdchto bunk\u00e1ch (obr\u00e1zok 4)<\/strong>.<\/p>\n

\u010cast\u00fdm n\u00e1lezom s\u00fa fibr\u00ednov\u00e9 mikrotromby v p\u013e\u00facnych kapil\u00e1rach aj vo v\u00e4\u010d\u0161\u00edch cievach(10) (obr\u00e1zok 3B)<\/strong>. Mikrotromby nie s\u00fa raritou ani pri in\u00fdch v\u00edrusov\u00fdch pneum\u00f3ni\u00e1ch, ako to bolo napr\u00edklad pri SARS, omnoho zriedkavej\u0161ie pri chr\u00edpke(11,14). V pr\u00edpade COVID-19 je ich v\u00fdskyt je prekvapivo \u010dast\u00fd aj u pacientov, ktor\u00ed dost\u00e1vali antikoagula\u010dn\u00fa terapiu(14). Endotelov\u00e9 bunky s\u00fa \u010dasto napu\u010dan\u00e9 a\u017e s tvorbou vakuol a so zdvojen\u00edm baz\u00e1lnej membr\u00e1ny. V niektor\u00fdch pr\u00edpadoch nach\u00e1dzame kapil\u00e1rne po\u0161kodenie s nekr\u00f3zou kapil\u00e1r, prolifer\u00e1ciou kapil\u00e1r a z\u00e1palovou infiltr\u00e1ciou(10).<\/p>\n

Pre proliferat\u00edvnu f\u00e1zu je typick\u00e9 pretrv\u00e1vanie reakt\u00edvnej hyperpl\u00e1zie pneumocytov II. typu a prolifer\u00e1cia fibroblastov sprev\u00e1dzan\u00e1 expanziou stien alveolov s n\u00e1sledn\u00fdm prestupom do alveol(3-6,10) (obr\u00e1zok 3D)<\/strong>. M\u00f4\u017ee by\u0165 pr\u00edtomn\u00e1 aj skvam\u00f3zna metapl\u00e1zia(5,11). \u010cast\u00fdmi n\u00e1lezmi, ktor\u00e9 predstavuj\u00fa komplik\u00e1cie DAD s\u00fa fibr\u00f3za p\u013e\u00fac a sekund\u00e1rna bakteri\u00e1lna, menej \u010dasto hubov\u00e1 infekcia(3,10).<\/p>\n

Distrib\u00facia DAD vo v\u0161etk\u00fdch f\u00e1zach v pr\u00edpade COVID-19 nie je uniformn\u00e1, s\u00fa\u010dasne nach\u00e1dzame zmeny v r\u00f4znych \u0161t\u00e1di\u00e1ch, hlavne v stredn\u00fdch a doln\u00fdch lalokoch p\u013e\u00fac(5). Lo\u017eisk\u00e1 postihnut\u00e9ho parench\u00fdmu s\u00fa rozlo\u017een\u00e9 nerovnomerne(10). Polak a spol. rozdelili n\u00e1lez pri COVID-19 pneum\u00f3nii odli\u0161ne, a to na 3 hlavn\u00e9 histologick\u00e9 vzory: 1. epitelov\u00fd s reakt\u00edvnymi zmenami epitelu, pr\u00edtomn\u00fd u 85 % pacientov, 2. vaskul\u00e1rny s mikrovaskul\u00e1rnym po\u0161koden\u00edm, mikrotrombami a ak\u00fatnou fibrin\u00f3znou a organizuj\u00facou sa pneum\u00f3niou (59 % pacientov) a 3. fibrotick\u00fd s fibr\u00f3zou interst\u00edcia (22 % pacientov). Jednotliv\u00e9 vzory sa aj prekr\u00fdvali, pri\u010dom fibr\u00f3za sa vyskytuje od 3. t\u00fd\u017ed\u0148a(4).<\/p>\n

Existuj\u00fa ind\u00edcie, \u017ee COVID-19 m\u00f4\u017ee minim\u00e1lne u \u010dasti pacientov ako prim\u00e1rnu odpove\u010f vyvol\u00e1va\u0165 organizuj\u00facu sa pneum\u00f3niu, pri\u010dom k rozvoju znakov DAD doch\u00e1dza a\u017e nesk\u00f4r v d\u00f4sledku in\u00fdch udalost\u00ed, napr\u00edklad p\u013e\u00facnej ventil\u00e1cie alebo trombotickej mikroangiopatie(7,10). Organizuj\u00faca sa pneum\u00f3nia vyvolan\u00e1 v\u00edrusmi bola op\u00edsan\u00e1 v minulosti napr\u00edklad v s\u00favislosti so SARS, MERS aj H1N1(7,12). Na presk\u00famanie tejto mo\u017enosti v pr\u00edpade COVID-19 v\u0161ak nie je dostupn\u00e9 dostato\u010dn\u00e9 mno\u017estvo pr\u00e1c opisuj\u00facich n\u00e1lez na p\u013e\u00facach vo ve\u013emi skorom \u0161t\u00e1diu ochorenia(7).<\/p>\n

Z na\u0161ich pozorovan\u00ed pri anal\u00fdze po\u010detn\u00fdch pitvan\u00fdch pr\u00edpadov \u00famrt\u00ed na COVID-19 m\u00f4\u017eeme pri mnoh\u00fdch kon\u0161tatova\u0165 pr\u00edtomnos\u0165 v\u0161etk\u00fdch op\u00edsan\u00fdch histologick\u00fdch n\u00e1lezov, nezriedka s\u00fa\u010dasne na rozli\u010dn\u00fdch miestach p\u013e\u00fac. T\u00e1to skuto\u010dnos\u0165 poukazuje na n\u00e1stup infekcie v r\u00f4znych \u010dastiach p\u013e\u00fac s odli\u0161n\u00fdm \u010dasov\u00fdm odstupom. Tak\u00e9to paralelne sa vyskytuj\u00face n\u00e1lezy COVID-19 pneum\u00f3nie s\u00fa ove\u013ea \u010dastej\u0161ie u pacientov z 2. vlny pand\u00e9mie, na rozdiel od 1. vlny (na Slovensku do konca j\u00fana 2020), kde spravidla dominovalo mas\u00edvne a uniformn\u00e9 postihnutie cel\u00e9ho tkaniva p\u013e\u00fac.<\/p>\n

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N\u00e1lez v in\u00fdch org\u00e1noch<\/h3>\n

Maiese a spol. vo svojej preh\u013eadovej \u0161t\u00fadii analyzuj\u00facej histologick\u00fd a pitevn\u00fd n\u00e1lez v pr\u00e1cach publikovan\u00fdch do j\u00fana roku 2020 nena\u0161li d\u00f4kaz \u0161pecifick\u00fdch zmien v in\u00fdch org\u00e1noch okrem p\u013e\u00fac. Medzi mimop\u013e\u00facnymi n\u00e1lezmi dominuj\u00fa mikrotromby a endotelit\u00edda r\u00f4znych org\u00e1nov, \u0161okov\u00e9 zmeny pe\u010dene, obli\u010diek a \u010driev, hemofagocyt\u00f3za v kostnej dreni a slezine(3).<\/p>\n

Zrejme ani kardi\u00e1lne komplik\u00e1cie pri ochoren\u00ed COVID-19 nie s\u00fa sp\u00f4soben\u00e9 priamym po\u0161koden\u00edm myokardu v\u00edrusom(3,13). Medzi frekventovan\u00e9 zmeny na srdci patria mikrotromby pretrv\u00e1vaj\u00face aj po kl\u00edrense v\u00edrusu a amyloid\u00f3za srdca. Myokardit\u00edda m\u00e1va v pr\u00edpade n\u00e1lezu ve\u013emi limitovan\u00fd rozsah(13).<\/p>\n

\u010cast\u00e9 neurologick\u00e9 pr\u00edznaky pri ochoren\u00ed COVID-19 poukazuj\u00fa na mo\u017en\u00fd neurotropizmus v\u00edrusu. Imunohistochemicky s\u00fa vzorky nervov\u00e9ho tkaniva v drvivej v\u00e4\u010d\u0161ine negat\u00edvne, okrem pr\u00edpadu n\u00e1lezu pozitivity v nervus vagus, nervus glossopharyngeus a ojedinel\u00fdch buniek v pred\u013a\u017eenej mieche celkovo u 16 pacientov. Vo v\u0161eobecnosti medzi naj\u010dastej\u0161ie n\u00e1lezy patr\u00ed intrakrani\u00e1lne krv\u00e1canie a encefalomal\u00e1cie r\u00f4z- nej lokaliz\u00e1cie a rozsahu a mierny, fok\u00e1lny, T-lymfocytov\u00fd infiltr\u00e1t v okol\u00ed ciev, v parench\u00fdme mozgu a leptomeningoch(16).<\/p>\n

S v\u00fdnimkou p\u013e\u00fac boli v\u00edrusov\u00e9 partikuly dok\u00e1zan\u00e9 najm\u00e4 v obli\u010dk\u00e1ch a \u010dreve(3,14). U pacientov s potvrden\u00fdm ochoren\u00edm COVID-19 s indikovan\u00fdm bioptick\u00fdm vy\u0161etren\u00edm boli naj\u010dastej\u0161\u00edmi n\u00e1lezmi ak\u00fatne tubul\u00e1rne po\u0161kodenie, fok\u00e1lna segment\u00e1lna glomeruloskler\u00f3za, po\u0161kodenie endotelu a trombotick\u00e1 mikroangiopatia(15). \u010cast\u00e9 gastrointestin\u00e1lne \u0165a\u017ekosti nekoreluj\u00fa s hor\u0161ou progn\u00f3zou pacientov. Mikroskopicky je pr\u00edtomn\u00e9 po\u0161kodenie endotelu ciev submuk\u00f3zy, lymfo-plazmocytov\u00fd z\u00e1palov\u00fd infiltr\u00e1t a ed\u00e9m lamina propria(14). V pr\u00edpade zn\u00e1mok po\u0161kodenia pe\u010dene existuj\u00fa ind\u00edcie, \u017ee u \u010dasti pacientov m\u00f4\u017eu by\u0165 zmeny sp\u00f4soben\u00e9 priamou infekciou v\u00edrusom(18).<\/p>\n

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Imunohistochemick\u00e1 detekcia SARS-CoV-2 infekcie<\/h3>\n

Mikroskopick\u00fd obraz v pr\u00edpade v\u00edrusov\u00fdch pneum\u00f3ni\u00ed vr\u00e1tane COVID-19 spravidla nie je dostato\u010dne \u0161pecifick\u00fd. V mnoh\u00fdch pr\u00edpadoch je potrebn\u00e9 etiologick\u00fd agens v nekroptick\u00fdch a bioptick\u00fdch vzork\u00e1ch dok\u00e1za\u0165 met\u00f3dami ako PCR, in situ hybridiz\u00e1cia, imunofluorescen\u010dn\u00fdmi met\u00f3dami alebo<\/p>\n

imunohistochemicky (IHC)(6). Najjednoduch\u0161ia a najlacnej\u0161ia je rutinne vyu\u017e\u00edvan\u00e1 imunohistoch\u00e9mia.<\/p>\n

V pr\u00edpade imunohistochemickej diagnostiky COVID-19 vznik\u00e1 nieko\u013eko probl\u00e9mov. Najz\u00e1va\u017enej\u0161\u00ed je, \u017ee IHC detekcia v\u00edrusu v p\u013e\u00facnom tkanive je spo\u013eahliv\u00e1 len v priebehu prv\u00fdch 2 t\u00fd\u017ed\u0148ov ochorenia, vz\u00e1cne mo\u017eno v\u00edrus dok\u00e1za\u0165 e\u0161te do 4 t\u00fd\u017ed\u0148ov(10). In\u00fdm z\u00e1va\u017en\u00fdm probl\u00e9mom v pr\u00edpade IHC d\u00f4kazu pr\u00edtomnosti SARS-CoV-2 vo vy\u0161etrovanom tkanive je \u010dasov\u00fd odstup vykonania pitvy po \u00famrt\u00ed. Na\u0161e sk\u00fasenosti ukazuj\u00fa, \u017ee imunohistochemick\u00e1 reaktivita zost\u00e1va zachovan\u00e1 pribli\u017ene do 30 hod\u00edn po smrti. Po tomto obdob\u00ed sa imunohistochemick\u00e1 detekcia v\u00edrusu st\u00e1va nespo\u013eahlivou a so zv\u00e4\u010d\u0161uj\u00facim sa \u010dasov\u00fdm intervalom v\u00fdkonu pitvy od \u00famrtia a\u017e nemo\u017enou. Dostupn\u00e9 pr\u00e1ce v\u0161ak v\u00e4\u010d\u0161inou \u010das pitvy neuv\u00e1dzaj\u00fa, preto nie je mo\u017en\u00e9 vyjadri\u0165 sa k zistenej spo\u013eahlivosti IHC(17).<\/p>\n

\u010eal\u0161ie \u0165a\u017ekosti vypl\u00fdvaj\u00fa z existencie mno\u017estva komer\u010dne dostupn\u00fdch, ale nedostato\u010dne overen\u00fdch protil\u00e1tok, \u010dasto s vysokou skr\u00ed\u017eenou reaktivitou v imunohistochemickej aplik\u00e1cii. Za pou\u017eitia t\u00fdchto protil\u00e1tok bolo v roku 2020 publikovan\u00fdch mno\u017estvo pr\u00e1c s \u00fadajn\u00fdm n\u00e1lezom priameho v\u00edruso- v\u00e9ho po\u0161kodenia r\u00f4znych tkan\u00edv.<\/p>\n

Z tohto d\u00f4vodu sme testovali protil\u00e1tky na tkanivov\u00fdch vzork\u00e1ch p\u013e\u00fac pacientov zosnul\u00fdch na n\u00e1sledky COVID-19 a na tkaniv\u00e1ch z obdobia pred r. 2020. Testovali sme na tkanivov\u00fdch rezoch bez antig\u00e9novej revitaliz\u00e1cie a s revitaliz\u00e1ciou 30 min. v citr\u00e1tovom pufri pH 5 alebo TRIS pufri pH 8, pri teplote varu. Negat\u00edvne kontroly boli jednak prepar\u00e1ty inkubovan\u00e9 bez prim\u00e1rnej protil\u00e1tky alebo inkubovan\u00e9 s my\u0161ac\u00edm alebo kr\u00e1li\u010d\u00edm s\u00e9rom pri rovnakom rieden\u00ed ako testovan\u00e1 protil\u00e1tka. S cie\u013eom dosiahnutia vy\u0161\u0161ej \u0161pecificity sme protil\u00e1tky testovali s inkub\u00e1ciou cez noc pri zv\u00fd\u0161en\u00fdch riedeniach. Okrem skr\u00ed\u017eenej reaktivity niektor\u00fdch protil\u00e1tok v tkaniv\u00e1ch z\u00edskan\u00fdch roky pred objaven\u00edm sa SARS-CoV-2 v\u00edrusu, viacer\u00e9 protil\u00e1tky sa uk\u00e1zali ako nevhodn\u00e9 pre imunohistochemick\u00fa aplik\u00e1ciu (tabu\u013eka 1)<\/strong>. V p\u013e\u00facnom tkanive sa uk\u00e1zala spo\u013eahliv\u00e1 imunohistochemick\u00e1 detekcia SARS-CoV-2 v\u00edrusu predov\u0161etk\u00fdm v skorom \u0161t\u00e1diu infekcie, v bronchiol\u00e1rnom, bronch\u00e1lnom epiteli a najm\u00e4 v odlupuj\u00facich sa v\u00edrusom na- padnut\u00fdch alveol\u00e1rnych epit\u00e9li\u00e1ch (obr\u00e1zok 4)<\/strong>. Zriedkavej\u0161ie sme pozorovali pozitivitu aj v hyperplastick\u00fdch alebo syncyci\u00e1lnych obrovsk\u00fdch alveol\u00e1rnych epitelov\u00fdch bunk\u00e1ch. Celkom ojedinelo bola pozitivita v cytoplazme ve\u013ek\u00fdch buniek s ve\u013ek\u00fdm mech\u00farikov\u00fdm jadrom a ve\u013ek\u00fdm jadierkom, pravdepodobne s v\u00edrusov\u00fdmi cytopatick\u00fdmi znakmi (obr\u00e1zok 4)<\/strong>.<\/p>\n

 <\/p>\n

Z\u00e1ver<\/span><\/h3>\n

Pand\u00e9mia sp\u00f4soben\u00e1 v\u00edrusom SARS-CoV-2, s n\u00e1hlym n\u00e1stupom a r\u00fdchlym roz\u0161\u00edren\u00edm po celej zemeguli, odhalila potrebu \u010do najr\u00fdchlej\u0161ieho a spo\u013eahliv\u00e9ho objasnenia patologickej podstaty ochorenia. Do popredia sa tak po rokoch zazn\u00e1vania a\u017e odmietania patologickoanatomickej pitvy dost\u00e1va posmrtn\u00e1 anal\u00fdza zosnul\u00fdch, ktor\u00e1 je zdrojom z\u00e1kladn\u00fdch inform\u00e1ci\u00ed o patogen\u00e9ze ochorenia a rozsahu po\u0161kodenia v r\u00f4znych tkaniv\u00e1ch. Anal\u00fdza pitevn\u00fdch n\u00e1lezov poskytla z\u00e1kladn\u00e9 poznatky o chorobe COVID-19 a prispela okrem pochopenia patol\u00f3gie aj k smerovaniu terapeutick\u00fdch strat\u00e9gi\u00ed pri lie\u010dbe tohto z\u00e1va\u017en\u00e9ho ochorenia. Imunohistochemick\u00fd d\u00f4kaz pr\u00edtomnosti infekcie SARS-CoV-2 vo vy\u0161etrovanom tkanive je r\u00fdchla a finan\u010dne nen\u00e1ro\u010dn\u00e1 met\u00f3da. Je v\u0161ak limitovan\u00e1 \u010dasov\u00fdm odstupom pitvy po smrti a dostupnos\u0165ou protil\u00e1tok s vysokou \u0161pecificitou a senzitivitou.<\/p>\n

 <\/p>\n

Oznam<\/em><\/strong><\/p>\n

T\u00e1to pr\u00e1ca bola podporen\u00e1 Agent\u00farou na podporu v\u00fdskumu a v\u00fdvoja na z\u00e1klade Zmluvy \u010d. PP-COVID-20-0051.<\/em><\/p>\n

 <\/p>\n

LITERAT\u00daRA<\/strong><\/p>\n

    \n
  1. WHO Coronavirus (COVID-19) Covid19.who.int. https:\/\/ covid19.who.int\/?gclid=CjwKCAiAm-2BBhANEiwAe7eyFK6TAzBmS- jQHhnOf7d3BMDSrF6vSN35KMeJRQnlWh_tG0uDaRpRhoCd7wQAvD_ BwE. Published 2021. Accessed April 8, 2021.<\/li>\n
  2. Gao Y, Ding M, Dong X, et al. Risk factors for severe and critically ill COVID\u201019 patients: A review. Allergy<\/em>. 2020;76(2):428-455. doi:10.1111\/ 14657<\/li>\n
  3. Maiese A, Manetti A, La Russa R, et al. Autopsy findings in COV- ID-19-related deaths: a literature review. Forensic Science, Medicine and <\/em>Pathology<\/em>. doi:10.1007\/s12024-020-00310-8<\/li>\n
  4. Polak S, Van Gool I, Cohen D, von der Th\u00fcsen J, van Paassen A systematic review of pathological findings in COVID-19: a pathophysio- logical timeline and possible mechanisms of disease progression. Modern Pathology<\/em>. 2020;33(11):2128-2138. doi:10.1038\/s41379-020-0603-3<\/li>\n
  5. Sessa F, Bertozzi G, Cipolloni L, et al. Clinical-Forensic Autopsy Findings to Defeat COVID-19 Disease: A Literature J Clin Med<\/em>. 2020;9(7):2026. doi:10.3390\/jcm9072026<\/li>\n
  6. Suster S, Moran Biopsy Interpretation Of The Lung<\/em>. 1st ed. Philadel- phia: Lippincott Williams & Wilkins; 2013:155-173.<\/li>\n
  7. Kory P, Kanne SARS-CoV-2 organising pneumonia: \u2018Has there been a widespread failure to identify and treat this prevalent condition in COV- ID-19?\u2019. BMJ Open Respir Res<\/em>. 2020;7(1):e000724. doi:10.1136\/bm- jresp-2020-000724<\/li>\n
  8. Al Nemer A. Histopathologic and Autopsy Findings in Patients Di- agnosed With Coronavirus Disease 2019 (COVID-19): What We Know So Far Based on Correlation With Clinical, Morphologic and Pathobi- ological Aspects. Adv Anat Pathol<\/em>. 2020;27(6):363-370. doi:10.1097\/ 0000000000000276<\/li>\n
  9. Salerno M, Sessa F, Piscopo A, et al. No Autopsies on COVID-19 Deaths: A Missed Opportunity and the Lockdown of Science. J Clin Med<\/em>. 2020;9(5):1472. doi:10.3390\/jcm9051472<\/li>\n
  10. Borczuk A, Salvatore S, Seshan S, et COVID-19 pulmonary pathology: a multi-institutional autopsy cohort from Italy and New York City. Modern Pathology<\/em>. 2020;33(11):2156-2168. doi:10.1038\/s41379-020-00661-1<\/li>\n
  11. B\u00f6sm\u00fcller H, Matter M, Fend F, Tzankov A. The pulmonary pathology of COVID-19. Virchows Archiv<\/em>. 2021;478(1):137-150. doi:10.1007\/ s00428-021-03053-1<\/li>\n
  12. Shieh W, Hsiao C, Paddock C, et al. Immunohistochemical, in si- tu hybridization, and ultrastructural localization of SARS-associated coronavirus in lung of a fatal case of severe acute respiratory syndrome in Taiwan. Hum Pathol<\/em>. 2005;36(3):303-309. doi:10.1016\/j. 2004.11.006<\/li>\n
  13. Bois MC, Boire NA, Layman AJ, et al. COVID-19-Associated Nonocclu- sive Fibrin Microthrombi in the Heart. Circulation<\/em>. 2021;143(3):230-243. doi:10.1161\/CIRCULATIONAHA.120.050754<\/li>\n
  14. Gupta A, Madhavan MV, Sehgal K, et Extrapulmonary manifesta- tions of COVID-19. Nat Med<\/em>. 2020;26(7):1017-1032. doi:10.1038\/s41591- 020-0968-3<\/li>\n
  15. Akilesh S, Nast CC, Yamashita M, et al. Multicenter Clinicopathologic Correlation of Kidney Biopsies Performed in COVID-19 Patients Presenting With Acute Kidney Injury or Proteinuria. Am J Kidney Dis<\/em>. 2021;77(1):82- e1. doi:10.1053\/j.ajkd.2020.10.001<\/li>\n
  16. Mukerji SS, Solomon What can we learn from brain autop- sies in COVID-19?. Neurosci Lett<\/em>. 2021;742:135528. doi:10.1016\/j.neu- let.2020.135528<\/li>\n
  17. Roden AC, Vrana JA, Koepplin JW, et Comparison of In Situ Hybrid- ization, Immunohistochemistry and Reverse Transcription-Droplet Digital Polymerase Chain Reaction for Severe Acute Respiratory Syndrome Coro- navirus 2 (SARSCoV-2)-Testing in Tissue [published online ahead of print, 2021 Mar 15]. Arch Pathol Lab Med<\/em>. 2021;10.5858\/arpa.2021-0008-SA. doi:10.5858\/arpa.2021-0008-SA<\/li>\n
  18. Wang Y, Liu S, Liu H, et al. SARS-CoV-2 infection of the liver directly contributes to hepatic impairment in patients with COVID-19. J Hepatol<\/em>. 2020;73(4):807-816. doi:10.1016\/j.jhep.2020.05.002<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

    *A rare case of autochthonous human dirofilariasis with the manifestation of pseudotumor of the epididymis caused by helminth Dirofilaria repens   \u00davod Ochorenie COVID-19 sa za\u010dalo \u0161\u00edri\u0165 koncom roka 2019 z \u010d\u00ednskeho mesta Wu-chan v provincii Chu-pej. Ku koncu marca 2021 bolo celosvetovo zaznamenan\u00fdch viac ako 132 mili\u00f3nov potvrden\u00fdch pr\u00edpadov n\u00e1kazy, pri\u010dom na ochorenie COVID-19<\/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":[297],"tags":[1729,1727,763,1728],"class_list":["post-2247","post","type-post","status-publish","format-standard","hentry","category-pathology","tag-autopsy","tag-covid-19","tag-immunohistochemistry","tag-viral-pneumonia","typ_clanku-original-work"],"acf":{"abstrakt":"

    During the COVID-19 pandemic, a need to quickly and reliably elucidate the pathogenesis of the disease emerged. Autopsy findings play an irreplaceable role in this process. Considering the clinical manifestation, current articles focus mainly on changes in the lungs, less in the other organs. The most common findings include various stages of diffuse alveolar damage, reactive and reparatory changes, and unusually frequent findings associated with coagulation impairment. As in many other types of viral pneumonia, the morphological changes are not specific enough. Special methods must be used to confirm the pathogen. Immunohistochemistry is the most available method in routine practice. In COVID-19, its use is limited particularly by the duration of the disease, the time over 30 hours from death to the autopsy, and the availability of antibodies with sufficient specificity and sensitivity.<\/p>\n

    Keywords: <\/strong>COVID-19, viral pneumonia, autopsy, immunohistochemistry<\/p>\n","casopis":[{"ID":2233,"post_author":"7","post_date":"2021-09-19 22:32:27","post_date_gmt":"2021-09-19 20:32:27","post_content":"We\u2019re bringing you this year\u2019s 1st<\/sup> issue of newslab, the scientific journal<\/strong>\r\n\r\n \r\n

      \r\n \t
    • Genomic variability of virus Sars-CoV-2<\/li>\r\n \t
    • Contribution of immunohistochemistry to histomorphological diagnostics of COVID-19 pneumonia<\/li>\r\n \t
    • Detection of microsatellite instability in Lynch syndrome-associated tumours<\/li>\r\n \t
    • Comparison of platelet glycoprotein analysis in the wholeh blood and plasma rich in platelets<\/li>\r\n \t
    • Tspan1 expression in prostatic adenocarcinoma<\/li>\r\n<\/ul>","post_title":"newslab","post_excerpt":"","post_status":"publish","comment_status":"closed","ping_status":"closed","post_password":"","post_name":"newslab-8","to_ping":"","pinged":"","post_modified":"2021-09-19 22:36:56","post_modified_gmt":"2021-09-19 20:36:56","post_content_filtered":"","post_parent":0,"guid":"https:\/\/www.newslab.sk\/casopis\/newslab-8\/","menu_order":0,"post_type":"casopis","post_mime_type":"","comment_count":"0","filter":"raw"}],"strana":"12 - 16","upload_clanok":{"ID":2248,"id":2248,"title":"NEWSLAB 1-2021_Krivosikova","filename":"NEWSLAB-1-2021_Krivosikova.pdf","filesize":1815276,"url":"https:\/\/www.newslab.sk\/wp-content\/uploads\/2021\/09\/NEWSLAB-1-2021_Krivosikova.pdf","link":"https:\/\/www.newslab.sk\/en\/contribution-of-immunohistochemistry-to-histomorphological-diagnostics-of-covid-19-pneumonia\/newslab-1-2021_krivosikova\/","alt":"","author":"7","description":"","caption":"","name":"newslab-1-2021_krivosikova","status":"inherit","uploaded_to":2247,"date":"2021-09-20 05:46:16","modified":"2021-09-20 05:46:16","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\/2247","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=2247"}],"version-history":[{"count":0,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/2247\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/media?parent=2247"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/categories?post=2247"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/tags?post=2247"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}