Verheijen R, Wiik A, De Jong B A, Høier-Madsen M, Ullman S, Halberg P, Van Venrooij W J
Department of Biochemistry, University of Nijmegen, Netherlands.
J Immunol Methods. 1994 Mar 10;169(2):173-82. doi: 10.1016/0022-1759(94)90261-5.
In this study we report the detection of autoantibodies to the nucleolar U3- and Th(7-2) ribonucleoprotein (RNP) particles in sera from patients with connective tissue diseases. The method described employs radioactively labelled antisense U3- and Th RNA which are hybridized to immunoprecipitated U3- or Th RNA from a HeLa cell extract. Of the 66 sera that were screened with this method seven sera (11%) precipitated only Th RNP, 16 sera (24%) precipitated only U3 RNP and 4 sera (6%) precipitated both U3- and Th RNP. Both anti-U3 RNP and anti-Th RNP activity appeared to be mostly associated with scleroderma or scleroderma-associated diseases. Using this method we also showed that some of the Th RNP particles in a cell extract are associated with the La autoantigen. We conclude that for the identification of immunoprecipitated RNAs this method is very sensitive and provides unambiguous data.
在本研究中,我们报告了在结缔组织病患者血清中检测到针对核仁U3和Th(7-2)核糖核蛋白(RNP)颗粒的自身抗体。所描述的方法采用放射性标记的反义U3和Th RNA,它们与从HeLa细胞提取物中免疫沉淀的U3或Th RNA杂交。用该方法筛选的66份血清中,7份血清(11%)仅沉淀Th RNP,16份血清(24%)仅沉淀U3 RNP,4份血清(6%)同时沉淀U3和Th RNP。抗U3 RNP和抗Th RNP活性似乎大多与硬皮病或硬皮病相关疾病有关。使用该方法我们还表明,细胞提取物中的一些Th RNP颗粒与La自身抗原相关。我们得出结论,对于免疫沉淀RNA的鉴定,该方法非常灵敏,并能提供明确的数据。