Fabien N, Moreira A, Lavergne J P, Desbos A, Surgey P, Alves de Olivera C, Gonzalo P, Venot A, Bienvenu J, Perrier H, Reboud J P, Monier J C
Laboratoire d Auto-Immunite, CHLS-HCL, Pierre-Bénite, France.
J Autoimmun. 1999 Aug;13(1):103-10. doi: 10.1006/jaut.1999.0291.
The autoantibodies (aAbs) directed against the ribosomal P proteins (RPP aAbs) are known to react mainly against epitopes localized within the common C-terminal sequence of the three acidic ribosomal P proteins, P0, P1 and P2. In order to investigate the opportunity to select short recombinant peptides of this common C-terminal sequence to detect the RPP-aAbs, the location of the epitopes recognized by ribosomal proteins (RP) aAb(+)sera of systemic lupus erythematosus patients (SLE) was investigated. Immunoblotting and ELISA techniques using extracted or recombinant, entire or cleaved RPP showed that 55% of the RP aAbs were directed against the three ribosomal P0, P1, and P2 proteins. The epitopes recognized by the RPP aAbs are located not only within the C-terminal sequence common to the three proteins but also within the N-terminal sequence of the P2 or P1 protein. The other RP aAbs sera (45%) did not react with all three proteins but with some of them, and showed the following pattern: P0(+)P1(+); P1(+); P2(+); P0(+)and P1(+). They recognized epitopes located in the region of the C-terminal sequence of the protein but not common to the three proteins. In addition two out of the six monoclonal Abs produced by immunization of mice using the P1 protein did not react with the peptide N-65 or N-71 of the P2 protein or with the C-terminal sequence of the three proteins. In conclusion, this study showed that the RPP aAb in SLE patients are not only directed against epitopes within the C-terminal sequence shared by the three acidic ribosomal P proteins. In view of these data it seems necessary to be cautious in using only a C-terminal peptide of ribosomal P proteins in tests performed to detect RPP aAb in human sera.
已知针对核糖体P蛋白的自身抗体(aAbs)(核糖体P蛋白自身抗体,RPP aAbs)主要与三种酸性核糖体P蛋白P0、P1和P2共同C末端序列中的表位发生反应。为了研究选择该共同C末端序列的短重组肽来检测RPP - aAbs的可能性,对系统性红斑狼疮患者(SLE)核糖体蛋白(RP)aAb(+)血清所识别表位的位置进行了研究。使用提取的或重组的、完整的或切割的RPP进行免疫印迹和ELISA技术检测显示,55%的RP aAbs针对三种核糖体P0、P1和P2蛋白。RPP aAbs识别的表位不仅位于三种蛋白共有的C末端序列内,还位于P2或P1蛋白的N末端序列内。其他RP aAbs血清(45%)并非与所有三种蛋白都发生反应,而是与其中一些反应,呈现以下模式:P0(+)P1(+);P1(+);P2(+);P0(+)和P1(+)。它们识别位于蛋白C末端序列区域但并非三种蛋白共有的表位。此外,用P1蛋白免疫小鼠产生的六种单克隆抗体中有两种不与P2蛋白的肽N - 65或N - 71或三种蛋白的C末端序列发生反应。总之,本研究表明SLE患者中的RPP aAb不仅针对三种酸性核糖体P蛋白共有的C末端序列内的表位。鉴于这些数据,在检测人血清中RPP aAb的试验中仅使用核糖体P蛋白的C末端肽似乎有必要谨慎行事。