Molecular Biological Approach to Anti Nuclear Factors

针对系统性红斑狼疮(SLE)、干燥综合征(SS)等胶原病患者血清中常见的抗SSA/Ro和SSB/La抗体，采用免疫双扩散法(DI)、对流免疫电泳法(CIE)和免疫印迹法(IB)对几种材料的SSA/Ro和SSB/La抗原性进行了检测。IB法对检测这两种自身抗体具有足够的敏感性和特异性。抗SSA/Ro抗体主要与58Kd多肽反应，抗SSB/La抗体主要与42Kd和40Kd多肽反应。此外，我们还发现部分抗SSA/Ro抗血清能与64Kd多肽发生反应。同时，通过DEAE柱层析、Sephacryl柱层析和免疫亲和层析从人胸腺、兔胸腺丙酮粉和培养的人细胞系(KB细胞)中部分纯化了抗体。研究表明，自身免疫性疾病患者血清中SS-A/Ra和SS-B/La自身抗体的出现频率高于以往报道。

Anti SSA/Ro and SSB/La antibodies are frequently found in sera from patients with systemic lupus erythematosus (SLE), Sjogren's syndrome (SS) and other collagen diseases.Both SSA/Ro and SSB/La antigenicc properties of several meterials were examined by double immunodiffusion technique (DI),counterimmunoelectrophoresis(CIE),and immunoblotting(IB) method, using monospecific antisera. The IB method was enough sensitive and specific to detect both autoantibodies. The anti SSA/Ro antibody chiefly reactive with 58Kd peptide and anti SSB/La antibody reactive with 42Kd and 40Kd peptides on a nitrocellulose membrane. Furthermore, we found some of the anti SSA/Ro antisera could react with 64Kd peptide. This indicates a heterogeneity of the antibody.In the meanwhile, the antibens were partially purified by DEAE column chromatography,sephacryl chromatography and immunoaffinity chromatography from a human apleen,a rabbit thymus acetone powder and cultured human cell line (KB cells). It became proved that the SS-A/Ra and SS-B/La autoantibodies appeared more frequently in Sera from patients with autoimmune diseases than those of previous reports.