Fusconi M, Cassani F, Govoni M, Caselli A, Farabegoli F, Lenzi M, Ballardini G, Zauli D, Bianchi F B
Cattedra di Semeiotica Medica, University of Bologna, Italy.
Clin Exp Immunol. 1991 Feb;83(2):291-7. doi: 10.1111/j.1365-2249.1991.tb05630.x.
The specificities of anti-nuclear antibodies (ANA) reacting with multiple nuclear dots (MND-ANA) present in about 15% primary biliary cirrhosis sera were studied by Western blot analysis with nuclear fractions from a human cell line. Reactivity with two broad bands of 78-92 kD and 96-100 kD of the insoluble fraction was present exclusively in MND-ANA-positive sera. Antibodies eluted from these proteins specifically retained the immunofluorescence reactivity of MND-ANA. Immunomorphological analysis by a pre-embedding technique revealed that the antibody specifically binds to nuclear regions resembling in size and number nuclear bodies. Since these structures are absent in immature rate endometrial cell and can be induced by diethylstilbestrol, we tested MND-ANA by immunofluorescence on cryostat sections of uteri from hormone-treated and untreated immature rats. A strong reaction of nuclear dots was observed predominantly in endometrial cells of hormone treated rats. We thus conclude that MND-ANA present in primary biliary cirrhosis sera are directed against 78-92-kD and 96-100-kD nuclear proteins located in nuclear bodies.
采用来自人细胞系的细胞核组分进行蛋白质印迹分析,研究了约15%原发性胆汁性肝硬化血清中存在的与多个核点反应的抗核抗体(ANA)(MND-ANA)的特异性。仅在MND-ANA阳性血清中存在与不溶性组分中78 - 92 kD和96 - 100 kD两条宽条带的反应性。从这些蛋白质上洗脱的抗体特异性保留了MND-ANA的免疫荧光反应性。采用包埋前技术进行免疫形态学分析显示,该抗体特异性结合大小和数量与核体相似的核区域。由于这些结构在未成熟大鼠子宫内膜细胞中不存在,且可由己烯雌酚诱导产生,因此我们通过免疫荧光对激素处理和未处理的未成熟大鼠子宫冰冻切片进行了MND-ANA检测。主要在激素处理大鼠的子宫内膜细胞中观察到核点的强烈反应。因此我们得出结论,原发性胆汁性肝硬化血清中存在的MND-ANA针对位于核体中的78 - 92 kD和96 - 100 kD核蛋白。