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目的研究B淋巴细胞刺激因子(B lymphocyte stimulator,BlyS)、抗-BlyS(BlyS-Ab)及其特异性免疫复合物(BLyS-IC)在各种自身免疫病(AID)患者中的检出水平及其在AID中的作用。方法优化试验反应条件,建立检测BlyS、BlyS-Ab及BlyS-IC的酶联免疫吸附试验(ELISA)测定法,并进行方法学考核和临床检测。结果优化的ELISA对BlyS的最低检出量可达0.4μ／L,在3．2～400．0μg／L浓度范围内呈现良好的线性关系(r>0．99)。BlyS-Ab精密度平均变异系数(CV)为7．4％,特异阻断抑制试验最高抑制率达83．3％。在系统性红斑狼疮(SLE)、类风湿关节炎(RA)、混合性结缔组织病(MCTD)等AID及抗核抗体(ANA)阳性的其他内科疾病(包括肾小球肾炎、过敏性紫癜和呼吸系统疾病)患者血清中BlyS质量浓度、BlyS-Ab及BlyS-IC水平均明显高于正常对照组(P<0．01)。ANA滴度与患者血清BlyS及BlyS-Ab水平呈平行升高趋势,但未发现BlyS水平与ANA某一特定荧光核型有相关关系。结论成功建立了人血清BlyS、BlyS-Ab及BlyS-IC的ELISA测定法,该方法敏感、特异、可靠。在SLE、RA和MCTD等患者中均有高水平的BlyS,同时伴随ANA、BlyS- Ab及BlyS-IC的升高。BlyS-Ab和BlyS-IC可能有助于AID诊断,并对阐明AID发病机制有一定意义。
Objective To study the detection level of B lymphocyte stimulator (BlyS), anti-BlyS (BlyS-Ab) and its specific immune complex (BLyS-IC) in patients with various autoimmune diseases And its role in AID. Methods The reaction conditions were optimized. The enzyme-linked immunosorbent assay (ELISA) was established for detection of BlyS, BlyS-Ab and BlyS-IC, and the methodological test and clinical test were performed. Results The optimized ELISA showed that the detection limit of BlyS was 0.4μL and the linear range was 3.2-400.0μg / L (r> 0.99). The BlyS-Ab precision average coefficient of variation (CV) was 7.4%, and the highest inhibition rate by specific blocking inhibition test was 83.3%. In other medical diseases (including glomerulonephritis, Henoch-Schonlein purpura, and Henoch-Schonlein purpura) which are positive for AID and antinuclear antibody (ANA) such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), mixed connective tissue disease (MCTD) BlyS-Ab and BlyS-IC levels in patients with respiratory diseases were significantly higher than those in normal controls (P <0.01). The ANA titer showed a parallel trend with the serum levels of BlyS and BlyS-Ab. However, no correlation was found between the level of BlyS and a specific fluorescent karyotype of ANA. Conclusion The ELISA method of human serum BlyS, BlyS-Ab and BlyS-IC has been successfully established. The method is sensitive, specific and reliable. High levels of BlyS are present in patients with SLE, RA and MCTD, accompanied by an increase in ANA, BlyS-Ab, and BlyS-IC. BlyS-Ab and BlyS-IC may contribute to the diagnosis of AID, and to clarify the pathogenesis of AID has some significance.