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目的:不依赖抗原情况下,分别用TLRs的激动剂及细胞因子单独和联合应用,对不同人群永生化B细胞体外刺激及诱导抗体产生,观察在疾病状态下B细胞活化状态,为寻找自身免疫病更敏感的新指标奠定基础。方法:用EpsteinBarr病毒(EBV)攻击SLE、RA患者B细胞,使之成为永生化的B淋巴母细胞系,体外用CpG-ODN2006、LPS及细胞因子与细胞共培养,免疫散射比浊法测定刺激后不同时段的总IgG水平,进行重复测量设计的方差分析。结果:刺激后细胞上清抗体分泌量增高10倍左右;SLE和RA患者抗体分泌量高于正常人;刺激后18d抗体分泌效应差异最为明显;CpG-ODN2006&IL-4&IL-6三者联合的刺激最好;经诱导后的抗体,均未检测到疾病特异性的血清学指标。结论:在疾病状态下,体外给予刺激后,B细胞活化状态更高,但未诱导出疾病特异的自身抗体。
OBJECTIVE: Independent of antigens, the immortalized B cells of different populations were stimulated with antibodies and cytokines alone or in combination with TLRs to induce antibody production and to observe the activation of B cells in disease states. In order to find out the mechanism of autoimmunity The more sensitive new indicators of disease lay the foundation. METHODS: B cells from SLE and RA patients were challenged with Epstein Barr virus (EBV) to become immortalized B lymphoblastoid cell lines. CpG-ODN2006, LPS and cytokines were co-cultured with the cells in vitro and stimulated by immunostaining turbidimetry After different periods of total IgG levels, repeated measures designed to analyze the variance. Results: The secretion of antibody in supernatant increased about 10-fold after stimulation; the secretion of antibody in patients with SLE and RA was higher than that in normal people; the difference of antibody secretion was most obvious at 18 days after stimulation; the stimulation of CpG-ODN2006 & IL-4 & IL- Good; after the induced antibody, no disease-specific serological markers were detected. CONCLUSIONS: B cells activated more in the disease state but stimulated in vitro but did not induce disease-specific autoantibodies.