Aoki S, Yoshikawa K, Yokoyama T, Nonogaki T, Iwasaki S, Mitsui T, Niwa S
Department of Pathology, Aichi Medical University, Japan.
Ann Rheum Dis. 1996 Jun;55(6):363-9. doi: 10.1136/ard.55.6.363.
To study antibodies to Escherichia coli O:14, which expresses large amounts of enterobacterial common antigen (ECA), and their corresponding antigen molecules in serum and synovial fluid samples from patients with rheumatoid arthritis (RA).
Enzyme linked immunosorbent assay (ELISA) was used to measure antibodies to heat killed E coli O:14 in serum and synovial fluid samples from patients with RA and control subjects including healthy donors and patients with osteoarthritis. ELISA was also used to perform absorption analyses of antibodies to E coli O:14 with several enteric bacteria and their lipopolysaccharide (LPS). In addition, antigenic molecules reacting with E coli O:14 antibodies from patients with RA were examined using immunoblot analysis and N-terminal amino acid analysis.
Compared with control subjects, patients with RA showed significantly increased titres of antibodies against heat killed E coli O:14 in 33 of 83 serum samples (39.8%) and 38 of 58 joint fluid samples (65.5%). Absorption analyses with enteric bacteria and their LPS resulted in the reduction of antibody titres to heat killed E coli O:14 in serum and synovial fluid samples from the RA patients. In addition, immunoblot analysis of the samples from RA patients revealed not only a ladder-like banding pattern equivalent to ECA associated with LPS, but also two clear bands of bacterial outer membrane proteins of 35 kDa (Omp A) and 38 kDa (Omp C), having amino acid sequence homology with those of other Enterobacteriaceae.
These results suggest that some patients with RA are sensitised to antigens common to Enterobacteriaceae, and this may prove relevant to the future development of immunotherapy for RA. Furthermore, this sensitisation to antigens found commonly in Enterobacteriaceae may have a key role in the pathogenesis of human RA similar to that described previously in our animal model.
研究类风湿关节炎（RA）患者血清和滑液样本中针对表达大量肠杆菌共同抗原（ECA）的大肠杆菌O:14的抗体及其相应抗原分子。
采用酶联免疫吸附测定（ELISA）检测RA患者以及包括健康供体和骨关节炎患者在内的对照受试者血清和滑液样本中针对热灭活大肠杆菌O:14的抗体。ELISA还用于用几种肠道细菌及其脂多糖（LPS）对大肠杆菌O:14抗体进行吸收分析。此外，使用免疫印迹分析和N端氨基酸分析检测与RA患者大肠杆菌O:14抗体反应的抗原分子。
与对照受试者相比，83份血清样本中的33份（39.8%）和58份关节液样本中的38份（65.5%）RA患者针对热灭活大肠杆菌O:14的抗体滴度显著升高。用肠道细菌及其LPS进行吸收分析导致RA患者血清和滑液样本中针对热灭活大肠杆菌O:14的抗体滴度降低。此外，对RA患者样本的免疫印迹分析不仅显示出与LPS相关的等同于ECA的梯状条带模式，还显示出两条清晰的35 kDa（Omp A）和38 kDa（Omp C）细菌外膜蛋白条带，其氨基酸序列与其他肠杆菌科细菌的氨基酸序列具有同源性。
这些结果表明，一些RA患者对肠杆菌科共同抗原敏感，这可能与RA免疫治疗的未来发展相关。此外，这种对肠杆菌科常见抗原的敏感性可能在人类RA发病机制中起关键作用，类似于我们之前在动物模型中所描述的。
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