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pristane在BALB/c小鼠中诱导产生高滴度的抗-Su和抗nRNP/Sm自身抗体。基于单特异性人类自身免疫血清,通过抗原捕获ELISA进行定量分析。

Pristane induces high titers of anti-Su and anti-nRNP/Sm autoantibodies in BALB/c mice. Quantitation by antigen capture ELISAs based on monospecific human autoimmune sera.

作者信息

Satoh M, Treadwell E L, Reeves W H

机构信息

Department of Medicine, University of North Carolina, Chapel Hill 27599-7280, USA.

出版信息

J Immunol Methods. 1995 May 11;182(1):51-62. doi: 10.1016/0022-1759(95)00022-3.

Abstract

Autoantibodies to Su and anti-nRNP/Sm are common in human and murine systemic lupus erythematosus (SLE), and are also produced by BALB/c mice with SLE-like autoimmunity induced by pristane. Antigen capture ELISAs employing monospecific human autoimmune IgG were developed to quantitate the production of anti-Su and anti-nRNP/Sm autoantibodies in 77 sera from BALB/c mice with pristane-induced autoimmunity. The sensitivity and specificity of the anti-Su antigen capture ELISA were 100% compared with immunoprecipitation of 35S-labeled cellular proteins. All 16 immunoprecipitation positive sera were positive in the anti-nRNP/Sm antigen capture ELISA (100% sensitivity), whereas 55/61 immunoprecipitation negative sera were negative by ELISA (90% specificity). The 6/61 immunoprecipitation negative sera that were ELISA positive were probably true positives because subsequent sera obtained from the same mice were positive by both techniques. Thus, the antigen capture ELISA may be somewhat more sensitive than immunoprecipitation. The titers of anti-Su and anti-nRNP/Sm positive antibodies in the sera were as high as 1:25,000-1:250,000 by ELISA, suggesting that autoantibodies may be produced in pristane-primed BALB/c mice at levels comparable to those seen in spontaneous autoimmune disease. We conclude that antigen capture ELISAs based on human autoimmune sera were highly sensitive and specific for detecting murine anti-Su and anti-nRNP/Sm antibodies. This technique will be useful for quantitating antibodies in murine autoimmune disease models, since antigen capture ELISA avoids the use of denatured or recombinant antigens, permitting antibodies recognizing tertiary and quaternary structures to be detected.

摘要

针对Su以及抗nRNP/Sm的自身抗体在人类和小鼠系统性红斑狼疮(SLE)中很常见,并且也由注射了 pristane 诱导出类似SLE自身免疫性的BALB/c小鼠产生。利用单特异性人类自身免疫性IgG开发了抗原捕获ELISA,以定量检测77份来自注射了pristane诱导出自身免疫性的BALB/c小鼠血清中抗Su和抗nRNP/Sm自身抗体的产生情况。与对35S标记的细胞蛋白进行免疫沉淀相比,抗Su抗原捕获ELISA的敏感性和特异性均为100%。所有16份免疫沉淀阳性血清在抗nRNP/Sm抗原捕获ELISA中均为阳性(敏感性100%),而61份免疫沉淀阴性血清中有55份通过ELISA检测为阴性(特异性90%)。6份ELISA阳性但免疫沉淀阴性的血清可能为真正的阳性,因为随后从同一小鼠获得的血清通过两种技术检测均为阳性。因此,抗原捕获ELISA可能比免疫沉淀稍敏感一些。通过ELISA检测,血清中抗Su和抗nRNP/Sm阳性抗体的滴度高达1:25,000 - 1:250,000,这表明在注射了pristane的BALB/c小鼠中产生的自身抗体水平与在自发性自身免疫疾病中所见水平相当。我们得出结论,基于人类自身免疫血清的抗原捕获ELISA在检测小鼠抗Su和抗nRNP/Sm抗体方面具有高度敏感性和特异性。由于抗原捕获ELISA避免使用变性或重组抗原,从而能够检测识别三级和四级结构的抗体,因此该技术将有助于对小鼠自身免疫疾病模型中的抗体进行定量分析。

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