Pei Fei, Zhao Ming-hui, Wang Ying-yu, Zhang Ying, Wang Hai-yan
Department of Nephrology, Peking University First Hospital, Beijing 100034, China.
Zhonghua Yi Xue Za Zhi. 2004 Feb 2;84(3):194-8.
Novel anti-mesangial cell antibodies have been identified in sera of majority patients with lupus nephritis in our previous study. The aims of current study were to investigate the association of anti-mesangial cell antibodies and anti-DNA antibodies, and whether the target antigens were on mesangial cell membrane.
Affinity parified anti-DNA antibodies and sera with depletion of anti-DNA antibodies from three patients with lupus nephritis were used in Western-blot analysis to identify antigens in culture human glomerular mesangial cell line. Cell membrane from in vitro cultured human mesagial cell line was separated by gradient centrifugation technique, and the target antigens were identified by Western-blot analysis using 1% Triton X-100 solublized membrane protein as antigens and four known anti-mesangial cell antibody positive sera from patients with lupus nephritis as probes.
Soluble proteins from in vitro cultured human mesangial cells could be blotted by both affinity purified anti-DNA antibodies (molecular weight at 63,000, 91,000 and 125,000) and sera with depletion of anti-DNA antibodies (molecular weight at 74,000 and 36,000). Proteins with molecular weight at 101,000, 91,000, 74,000 and 31,000 in mesangial cell membrane could be blotted by the four known anti-mesangial cell antibody positive sera.
Some anti-mesangial cell antibodies were not associated with anti-DNA antibodies and anti-mesiangial cell antibodies could direct interact with antigens located on mesangial cell membrane. Anti-mesangial cell antibodies may play an important role in the pathogenesis of lupus nephritis.
在我们之前的研究中,已在大多数狼疮性肾炎患者的血清中鉴定出新型抗系膜细胞抗体。本研究的目的是调查抗系膜细胞抗体与抗DNA抗体之间的关联,以及靶抗原是否位于系膜细胞膜上。
使用亲和纯化的抗DNA抗体以及来自三名狼疮性肾炎患者的去除抗DNA抗体的血清进行蛋白质印迹分析,以鉴定培养的人肾小球系膜细胞系中的抗原。通过梯度离心技术分离体外培养的人系膜细胞系的细胞膜,以1% Triton X-100溶解的膜蛋白作为抗原,使用四份已知的狼疮性肾炎患者抗系膜细胞抗体阳性血清作为探针,通过蛋白质印迹分析鉴定靶抗原。
体外培养的人系膜细胞的可溶性蛋白可被亲和纯化的抗DNA抗体(分子量分别为63,000、91,000和125,000)以及去除抗DNA抗体的血清(分子量分别为74,000和36,000)印迹。系膜细胞膜上分子量为101,000、91,000、74,000和31,000的蛋白质可被四份已知的抗系膜细胞抗体阳性血清印迹。
一些抗系膜细胞抗体与抗DNA抗体无关,抗系膜细胞抗体可直接与位于系膜细胞膜上的抗原相互作用。抗系膜细胞抗体可能在狼疮性肾炎的发病机制中起重要作用。