Department of Rheumatology, Leiden University Medical Center (LUMC), Leiden, The Netherlands.
Ann Rheum Dis. 2013 Jun;72(6):1059-63. doi: 10.1136/annrheumdis-2012-202747. Epub 2013 Mar 13.
Anticitrullinated protein antibodies (ACPA) are one of the best predictors for the development of rheumatoid arthritis. Nonetheless, relatively little information is present on the absolute concentration of ACPA in relation to total immunoglobulin (Ig) concentrations. Such information would be of relevance to compare ACPA levels to other antibody levels. Here, we estimated the relative abundance of ACPA Ig in serum and synovial fluid using a quantitative approach.
ACPA were purified using HiTrap Streptavidin columns coupled with biotinylated cyclic citrullinated peptide (CCP2). Total Ig and anti-CCP2 isotype reactivities were measured by ELISA.
ACPA were successfully isolated as substantial antibody amounts were eluted from sera of ACPA-positive patients and neglectable antibody amounts were eluted from sera of ACPA-negative patients. Up to 1 in 80 IgG-molecules were estimated to be ACPA. Strikingly, IgM-ACPA was most abundant in synovial fluid (with the highest enrichment in the range of one IgM-ACPA for every eight IgM-antibodies).
ACPA-IgG levels are estimated to be within the range of peak levels of protective antibody responses against recall antigens. IgM-ACPA is abundantly present in synovial fluid, suggesting the presence of a continuous ongoing autoimmune response in the synovial compartment.
抗瓜氨酸化蛋白抗体(ACPA)是类风湿关节炎发生的最佳预测因子之一。尽管如此,关于 ACPA 相对于总免疫球蛋白(Ig)浓度的绝对浓度的信息相对较少。此类信息对于将 ACPA 水平与其他抗体水平进行比较很重要。在这里,我们使用定量方法来估计血清和滑液中 ACPA-Ig 的相对丰度。
使用与生物素化环瓜氨酸肽(CCP2)偶联的 HiTrap Streptavidin 柱来纯化 ACPA。通过 ELISA 测量总 Ig 和抗 CCP2 同种型反应性。
ACPA 可成功分离,因为从 ACPA 阳性患者的血清中洗脱了大量的抗体,而从 ACPA 阴性患者的血清中洗脱了微不足道的抗体。估计多达 1/80 的 IgG 分子是 ACPA。值得注意的是,IgM-ACPA 在滑液中最为丰富(在每 8 个 IgM 抗体中就有一个 IgM-ACPA 的最高富集范围内)。
ACPA-IgG 水平估计处于针对回忆抗原的保护性抗体反应的峰值水平范围内。IgM-ACPA 在滑液中大量存在,表明在滑液隔室中存在持续进行的自身免疫反应。