自身免疫性疾病患者血清BlyS、抗-BlyS及其特异性免疫复合物的测定研究

Serum BlyS, anti-BlyS and BlyS-immune complex determination in patients with autoimmune diseases

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 investigate the prevalence of B lymphocyte stimulator（BlyS）, anti-BlyS （BlyS-Ab） and its specific immune complex（BlyS-IC） in patients with autoimmune diseases （AID）, and explores the role of the above items in pathogenesis of these diseases. Methods By optimizing experimental conditions, we had established three types of ELISA methods for serum levels of BlyS, BlyS-Ab and BlyS- IC, followed by assessing their methodology and serving them as a tool for clinical research. Results The lower detection limit of ELISA method for BlyS was determined to be at 0. 4 μg/L At a concentration of 3.2 - 400. 0 μg/L, there was a good learner correlations for BlyS （ r 〉 0. 99 ）. The intra-assay precision, namely average of coefficient values of BlyS-Ab was 7.4% , and the specific inhibition rate for BlyS-Ab by series concentration of BlyS was up to 83.3%. Clinical investigation revealed that serum levels of BlyS, BlyS-Ab and BlyS-IC were significantly higher in patients with AID including SLE, RA, MCTD and other medicine cases with positive ANA （i. e. nephritis, allergic purpurics and pulmonary disease etc. ） than that in normal control （In comparison with titers of ANA, serum BlyS and BlyS-Ab level were parallel increases）, but no correlation was found in this investigation between serum BlyS levels and certain fluorescent nuclear-type of ANA. Conclusions Detection methods for human serum BlyS, BlyS-Ab and BlyS-IC are successfully established. The assay results were specific, sensitive and reliable. Higher levels of serum BlyS were found in patients with multifarious AID, at the sametime, serum levels of ANA, BlyS-Ab as well as BlyS-IC are also increased. Therefore, this investigation is potentially helpful to diagnose AID, and plays a significant role to illustrate pathogenesis of these diseases.