副肿瘤性天疱疮伴发Castleman肿瘤分泌致病相关抗体的初步研究

A Preliminary Study on the Antibody Secreted in Castleman's Disease Associated with Paraneoplastic Pemphigus

目的Castleman肿瘤组织细胞培养及培养细胞上清液中抗体成分性质的鉴定。方法分离副肿瘤性天疱疮患者的肿瘤淋巴细胞进行培养,将培养细胞上清液超滤离心获得浓缩的IgG抗体为一抗,分别以鼠膀胱冰冻切片为底物行间接免疫荧光检查,以正常皮肤蛋白提取物为底物进行免疫印迹试验。RT-PCR,克隆测序分析培养肿瘤B细胞免疫球蛋白重链基因并与其他6例该病患者的基因相比较。结果培养细胞上清液中的IgG抗体能够在鼠膀胱上皮细胞间沉积;并且这些抗体能够识别皮肤中分子量分别为210000和190000的两种抗原成分。扩增培养肿瘤B细胞免疫球蛋白重链基因得到单一锐利条带,克隆测序发现该基因与另外6例患者的基因序列具有高度一致性。结论患者肿瘤组织能够分泌抗体,并且分泌抗体的性质与血液中抗体性质基本相同。

Objective To identify features of antibodies in the supernatants of cultured Castleman's disease cells.Methods Lymphocytes of Castleman's disease were isolated and cultured.Immunofluorescence and immunoblot assays were performed with IgG extracted from culture supernatants.The immunoglobulin heavy chaingene of cultured tumor B cells was analyzed by RT-PCR,cloning and sequencing.ResultsIg Gextracted from culture supernatant scouldattachtotheepithelialcellsurfacesofmousebladdertissues.Theantibodycouldalsoidentifytwoantigencomponents,210000and190000,ofnormalhumanepidermaltis-sues.ThesequencesimilaritywasfoundinimmunoglobulinheavychaingeneofculturedtumorBcellscom-paredwiththatof6patientswithCastleman'sdiseasepreviouslyreported.Conclusions Castleman's tumor associated with paraneoplastic pemphigus can secret autoantibody with similar features to that found in patients'sera.