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使用腺病毒感染的HeLa细胞检测低滴度自身抗体。

The use of adenovirus-infected HeLa cells for the detection of low titer autoantibodies.

作者信息

Slobbe R, Van Esch B, Kveder T, Van Venrooij W J

机构信息

Department of Biochemistry, University of Nijmegen, The Netherlands.

出版信息

J Immunol Methods. 1991 Apr 25;138(2):237-44. doi: 10.1016/0022-1759(91)90172-c.

Abstract

Following infection of HeLa cells with adenovirus type 5 the cellular La protein becomes predominantly associated with the virally encoded RNA polymerase III products VAI, and VAII, while most of the host RNA polymerase II (e.g. U1, U2, U4, U5 and mRNA) and RNA polymerase III transcription (e.g. U6 and pre-tRNAs) ceases. Other RNA polymerase III products such as the cellular Ro RNAs continue to be transcribed and assembled into ribonucleoprotein complexes containing the Ro (SS-A) antigens. Using a 32P-pulse chase-labeled, adenovirus-infected HeLa cellular extract as a source of antigen, anti-La (SS-B) and anti-Ro (SS-A) antibodies can be detected simultaneously using an immunoprecipitation assay. In the present study this method was found to be more sensitive in detecting anti-La antibodies then counter immunoelectrophoresis and immunoblotting. In studies of sera from patients suffering from rheumatic diseases the percentage positive for anti-La antibody was significantly elevated using this method, especially in patients with systemic lupus erythematosus.

摘要

用5型腺病毒感染HeLa细胞后,细胞内的La蛋白主要与病毒编码的RNA聚合酶III产物VAI和VAII结合,而大多数宿主RNA聚合酶II(如U1、U2、U4、U5和mRNA)以及RNA聚合酶III转录(如U6和前体tRNA)停止。其他RNA聚合酶III产物,如细胞内的Ro RNA,继续转录并组装成含有Ro(SS-A)抗原的核糖核蛋白复合物。使用经32P脉冲追踪标记的、腺病毒感染的HeLa细胞提取物作为抗原来源,通过免疫沉淀试验可同时检测抗La(SS-B)和抗Ro(SS-A)抗体。在本研究中,发现该方法在检测抗La抗体方面比对流免疫电泳和免疫印迹更敏感。在对风湿性疾病患者血清的研究中,使用该方法抗La抗体阳性百分比显著升高,尤其是系统性红斑狼疮患者。

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