电子显微学报
電子顯微學報
전자현미학보
JOURNAL OF CHINESE ELECTRON MICROSCOPY SOCIETY
2009年
6期
548-555
,共8页
黄晓峰%刘彦仿%尹文%王爽%亢君君%李永强
黃曉峰%劉彥倣%尹文%王爽%亢君君%李永彊
황효봉%류언방%윤문%왕상%항군군%리영강
肾综合征出血热%免疫复合物%免疫组织化学%激光扫描共聚焦显微镜技术%超微结构
腎綜閤徵齣血熱%免疫複閤物%免疫組織化學%激光掃描共聚焦顯微鏡技術%超微結構
신종합정출혈열%면역복합물%면역조직화학%격광소묘공취초현미경기술%초미결구
hemorrhagic fever with renal syndrome%immune complex%immunohistochemistry%laser scanning confocal microscopy%ultrastructure
目的:为确定肾综合征出血热(HFRS)不溶性免疫复合物(IC)在发病中的作用.方法:本研究采用HFRS病毒抗原和相应抗体在体外制备成特异性的不溶性IC,经尾静脉注射入小鼠体内,而后应用免疫荧光组织化学方法结合激光扫描共聚焦显微镜和电子显微镜观察了病毒抗原及IC在各主要脏器的分布及其致病变作用.结果:IC注射后24 h.主要见于血管系统如肝血窦和肾小球毛细血管之中.IC注射72 h后,IC主要存在于肾小球基底膜部位以及血管系膜中,呈细小或粗大的颗粒状.肾小球毛细血管中红细胞明确阳性.某些血管腔中仍充满荧光物质,有些荧光物质位于足细胞的胞质内.在肾小球周围的部分肾小管上皮细胞可有IC,包括抗原和C3.不溶性IC注射后168 h(7d),大多数肾小球结构接近正常,几乎看不到特异性的荧光.电镜观察发现病变以肾小球为主,发现系膜细胞伸出明显的胞质突起,并延伸至内皮和基膜之间.系膜细胞也有增生,可见沿细胞膜分布的中等数量的吞饮小泡以及胞浆中小的溶酶体颗粒.免疫沉积物可出现在肾小球系膜基质区,为不规则形,常聚集成团块状,呈散在分布,有时可见于内皮下、上皮下和/或基膜内.肾小球基膜呈不规则延伸、增厚,呈花斑状,并形成裂隙.IC注射后24 h,某些毛细血管袢管腔中有血小板聚集.注射后72 h,血小板聚集已不明显.某些毛细血管内皮细胞损伤严重,内皮细胞肿胀,部分区域内皮细胞突起脱落,基膜裸露.内皮细胞伸出许多绒毛状突起,互相交织融合形成拱状结构.胞浆中游离核糖体和粗面内质网增多.有些内皮细胞的胞质形成泡状结构.毛细血管管腔中充满一些电子致密的沉积物,并且这些电子致密物可通过内皮缺损与肾小球毛细血管基膜物质相连.肾小球囊壁层基膜外侧裸露,有胶原纤维形成.足细胞肿胀,突起融合,胞质中有溶酶体和吞饮(吞噬)小泡,吞饮小泡中舍有电子致密物.肾小球病变虽然比较明显,但肾小管病变相对较轻,仅肾小球周围的部分肾小管(主要是近曲小管)可出现溶酶体增生.所有对照动物未见发病,组织学检查未见明显病变.免疫荧光组织化学染色对照为阴性,单纯注射PAP的动物,经显色后,发现阳性反应都位于血管系统之中,各器官实质细胞未见阳性反应.说明出血热IC引起的致病变作用是特异性的.结论:不溶性IC主要沉积于肾小球的内皮下、基膜内、上皮下和毛细血管系膜内.HFRS的发病中,肾小球肾炎的改变可能是不溶性IC引起的.
目的:為確定腎綜閤徵齣血熱(HFRS)不溶性免疫複閤物(IC)在髮病中的作用.方法:本研究採用HFRS病毒抗原和相應抗體在體外製備成特異性的不溶性IC,經尾靜脈註射入小鼠體內,而後應用免疫熒光組織化學方法結閤激光掃描共聚焦顯微鏡和電子顯微鏡觀察瞭病毒抗原及IC在各主要髒器的分佈及其緻病變作用.結果:IC註射後24 h.主要見于血管繫統如肝血竇和腎小毬毛細血管之中.IC註射72 h後,IC主要存在于腎小毬基底膜部位以及血管繫膜中,呈細小或粗大的顆粒狀.腎小毬毛細血管中紅細胞明確暘性.某些血管腔中仍充滿熒光物質,有些熒光物質位于足細胞的胞質內.在腎小毬週圍的部分腎小管上皮細胞可有IC,包括抗原和C3.不溶性IC註射後168 h(7d),大多數腎小毬結構接近正常,幾乎看不到特異性的熒光.電鏡觀察髮現病變以腎小毬為主,髮現繫膜細胞伸齣明顯的胞質突起,併延伸至內皮和基膜之間.繫膜細胞也有增生,可見沿細胞膜分佈的中等數量的吞飲小泡以及胞漿中小的溶酶體顆粒.免疫沉積物可齣現在腎小毬繫膜基質區,為不規則形,常聚集成糰塊狀,呈散在分佈,有時可見于內皮下、上皮下和/或基膜內.腎小毬基膜呈不規則延伸、增厚,呈花斑狀,併形成裂隙.IC註射後24 h,某些毛細血管袢管腔中有血小闆聚集.註射後72 h,血小闆聚集已不明顯.某些毛細血管內皮細胞損傷嚴重,內皮細胞腫脹,部分區域內皮細胞突起脫落,基膜裸露.內皮細胞伸齣許多絨毛狀突起,互相交織融閤形成拱狀結構.胞漿中遊離覈糖體和粗麵內質網增多.有些內皮細胞的胞質形成泡狀結構.毛細血管管腔中充滿一些電子緻密的沉積物,併且這些電子緻密物可通過內皮缺損與腎小毬毛細血管基膜物質相連.腎小毬囊壁層基膜外側裸露,有膠原纖維形成.足細胞腫脹,突起融閤,胞質中有溶酶體和吞飲(吞噬)小泡,吞飲小泡中捨有電子緻密物.腎小毬病變雖然比較明顯,但腎小管病變相對較輕,僅腎小毬週圍的部分腎小管(主要是近麯小管)可齣現溶酶體增生.所有對照動物未見髮病,組織學檢查未見明顯病變.免疫熒光組織化學染色對照為陰性,單純註射PAP的動物,經顯色後,髮現暘性反應都位于血管繫統之中,各器官實質細胞未見暘性反應.說明齣血熱IC引起的緻病變作用是特異性的.結論:不溶性IC主要沉積于腎小毬的內皮下、基膜內、上皮下和毛細血管繫膜內.HFRS的髮病中,腎小毬腎炎的改變可能是不溶性IC引起的.
목적:위학정신종합정출혈열(HFRS)불용성면역복합물(IC)재발병중적작용.방법:본연구채용HFRS병독항원화상응항체재체외제비성특이성적불용성IC,경미정맥주사입소서체내,이후응용면역형광조직화학방법결합격광소묘공취초현미경화전자현미경관찰료병독항원급IC재각주요장기적분포급기치병변작용.결과:IC주사후24 h.주요견우혈관계통여간혈두화신소구모세혈관지중.IC주사72 h후,IC주요존재우신소구기저막부위이급혈관계막중,정세소혹조대적과립상.신소구모세혈관중홍세포명학양성.모사혈관강중잉충만형광물질,유사형광물질위우족세포적포질내.재신소구주위적부분신소관상피세포가유IC,포괄항원화C3.불용성IC주사후168 h(7d),대다수신소구결구접근정상,궤호간불도특이성적형광.전경관찰발현병변이신소구위주,발현계막세포신출명현적포질돌기,병연신지내피화기막지간.계막세포야유증생,가견연세포막분포적중등수량적탄음소포이급포장중소적용매체과립.면역침적물가출현재신소구계막기질구,위불규칙형,상취집성단괴상,정산재분포,유시가견우내피하、상피하화/혹기막내.신소구기막정불규칙연신、증후,정화반상,병형성렬극.IC주사후24 h,모사모세혈관번관강중유혈소판취집.주사후72 h,혈소판취집이불명현.모사모세혈관내피세포손상엄중,내피세포종창,부분구역내피세포돌기탈락,기막라로.내피세포신출허다융모상돌기,호상교직융합형성공상결구.포장중유리핵당체화조면내질망증다.유사내피세포적포질형성포상결구.모세혈관관강중충만일사전자치밀적침적물,병차저사전자치밀물가통과내피결손여신소구모세혈관기막물질상련.신소구낭벽층기막외측라로,유효원섬유형성.족세포종창,돌기융합,포질중유용매체화탄음(탄서)소포,탄음소포중사유전자치밀물.신소구병변수연비교명현,단신소관병변상대교경,부신소구주위적부분신소관(주요시근곡소관)가출현용매체증생.소유대조동물미견발병,조직학검사미견명현병변.면역형광조직화학염색대조위음성,단순주사PAP적동물,경현색후,발현양성반응도위우혈관계통지중,각기관실질세포미견양성반응.설명출혈열IC인기적치병변작용시특이성적.결론:불용성IC주요침적우신소구적내피하、기막내、상피하화모세혈관계막내.HFRS적발병중,신소구신염적개변가능시불용성IC인기적.
Objective: In order to determine the effect of the immunocomplexes on the pathogenesis of hemorrhagic fever with syndrome (HFRS) . Method:The insoluble antigen-antibody complexes was prepared in vitro, and was intravenously injected into BALB/c mice. The distribution of antigen, antibody and ICs was confirmed by fluorescence immunohistochemical method, combined laser scanning confocal microscopy and electron microscopy. Results: The results showed that IC was primarily distributed in blood vessels and sinusoids within 24 hours of the injection. From 72 hours after injection, the depositions of IC were found mainly on the basement membrane and in the spaces between mesangial cells in the kidney and on the wall of sinuses of the liver . The red blood cells within or without the blood vessels were IC positive. In the cytoplasm of podocytes and the tubular epithelia, IC fluorescence can be seen. Both the antigen, antibody and the C3 were foundin the same places. Under electron microscope, the glomeruli showed the main changes. The masangial cells proliferated with its processes extended between the endothelial cells and the hasement membrane. Deposits were found on the basement membranes, under the endothelia or under the epithelia. Irregular rarefied spaces could be seen in the deposits. Twenty-four hours after injection, blood platelets' aggregation within the glomerular capillaries could be discovered . After 72 hours, there were sloughing of endothelia with exposure of basement membrane. There was partial fusion of elongated villi to form arch structure. On the surfaces of red blood cells, there was evenly distributed electron dense particles. Conclusion: The results of this experiment showed clearly that: The insouble IC formed by antibody in excess, deposited principally on the basement membrane and was rarely seen in human cases.
