NEOANTIGEN OF THE POLYMERIZED 9TH COMPONENT OF COMPLEMENT - CHARACTERIZATION OF A MONOCLONAL-ANTIBODY AND IMMUNOHISTOCHEMICAL LOCALIZATION IN RENAL-DISEASE
NEOANTIGEN OF THE POLYMERIZED 9TH COMPONENT OF COMPLEMENT - CHARACTERIZATION OF A MONOCLONAL-ANTIBODY AND IMMUNOHISTOCHEMICAL LOCALIZATION IN RENAL-DISEASE
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DOI：
10.1172/jci111004
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发表时间：
1983-01-01
影响因子：
15.9
通讯作者：
MICHAEL, AF
中科院分区：
文献类型：
--
作者：
FALK, RJ;DALMASSO, AP;MICHAEL, AF
研制了抗人补体膜攻击复合体(MAC)新抗原C9[补体成分9]的单抗，并对其进行了鉴定。用间接免疫荧光显微镜观察这种新抗原在肾炎性和非肾炎性肾脏疾病中的分布。该抗体(Poly C9-MA)与补体激活血清中的一个在正常人血清(NHS)中检测不到的决定簇在酶联免疫吸附试验(ELISA)中发生反应。在NHS中孵育的酵母多糖颗粒与Poly C9-MA呈阳性免疫荧光染色，而在缺乏单个补体成分C3、C5、C6、C7、C8或C9的血清中孵育的酵母多糖颗粒与Poly C9-MA没有结合。用纯化的C9重建C9缺乏的血清，用Poly C9-MA恢复了荧光。Poly C9-MA与NHS处理的抗体包被的绵羊红细胞膜上的纯化的MC5b-9呈剂量依赖性阳性反应，但不与中间补体复合体反应。Poly C9-MA还与聚合C9(37度)在ELISA法和放射免疫分析中以剂量依赖的方式反应。C，h)(聚C9)，但不与单体C9共聚。增加未标记的聚C9或纯化的MC5b-9以相同的方式抑制125I-聚C9的RIA。显然，Poly C9-MA识别MAC和Poly C9共有的C9新抗原。免疫荧光显示Poly C9-MA在肾小球旁、系膜柄和血管壁与正常肾组织反应最弱。Poly C9-MA染色肾小球肾炎患者的肾组织，其模式与多克隆抗人C3相似。在非肾炎性肾病、糖尿病、高血压和梗阻性尿路病变患者的组织中，Poly C9-MA在系膜柄和肾小球旁区域、肾小管基底膜和血管壁呈强阳性反应。Poly C9-MA结合在晚期组织损伤区域尤为突出。Poly C9-MA经常在C3最少存在或不存在的位置染色。这些研究为补体激活提供了强有力的证据，不仅在肾炎性疾病中，而且在非肾炎性肾脏疾病中也是如此。
A monoclonal antibody to a neoantigen of the C9 [complement component 9] portion of the membrane attack complex (MAC) of human complement was developed and characterized. The distribution of this neoantigen was assessed by indirect immunofluorescence microscopy in nephritic and nonnephritic renal diseases. The antibody (Poly C9-MA) reacted on enzyme-linked immunosorbent assay (ELISA) with a determinant in complement-activated serum that was undetectable in normal human serum (NHS). Zymosan particles incubated in NHS had positive immunofluorescent staining with Poly C9-MA; binding of Poly C9-MA was not observed with zymosan particles incubated in sera deficient in individual complement components C3, C5, C6, C7, C8 or C9. Reconstitution of C9-deficient sera with purified C9 restored the fluorescence with Poly C9-MA. Poly C9-MA reacted positively by ELISA in a dose-dependent manner with purified MC5b-9 solubilized from membranes of antibody-coated sheep erythrocytes treated with NHS but not with intermediate complement complexes. Poly C9-MA also reacted in a dose-dependent manner on ELISA and in a radioimmunoassay with polymerized C9 (37.degree. C, 64 h) (poly C9) but not with monomeric C9. Increasing amounts of either unlabeled poly C9 or purified MC5b-9 inhibited the 125I-poly C9 RIA in an identical manner. Evidently, Poly C9-MA recognizes a neoantigen of C9 common to both the MAC and to poly C9. By immunofluorescence, Poly C9-MA reacted minimally with normal kidney tissue in juxtaglomerular loci, the mesangial stalk and vessel walls. Poly C9-MA stained kidney tissue from patients with glomerulonephritis in a pattern similar to that seen with polyclonal anti-human C3. In tissue from patients with nonnephritic renal disease, diabetes, hypertension and obstructive uropathy, Poly C9-MA was strongly reactive in the mesangial stalk and juxtaglomerular regions, tubular basement membranes and vascular walls. Poly C9-MA binding was especially prominent in areas of advanced tissue injury. Poly C9-MA frequently stained loci where C3 was either minimally present or absent. These studies provide strong evidence for complement activation not only in nephritic but also in nonnephritic renal diseases.