Krenn V, Vollmers H P, von Landenberg P, Schmausser B, Rupp M, Roggenkamp A, Müller-Hermelink H K
Institut für Pathologie, Universität Würzburg, Germany.
Virchows Arch. 1996 Feb;427(5):511-8. doi: 10.1007/BF00199512.
Several studies indicate a pathogenetic role of T-lymphocytes with specificity for heat shock proteins (HSP) in rheumatoid arthritis (RA). Surprisingly, there are no experimental data for B-lymphocytes with specificity for HSP. To investigate whether B-lymphocytes from rheumatoid synovial tissue show a specificity for HSP 60 we immortalized synovial tissue B-lymphocytes by the electrofusion technique and tested the specificity of the B-cell clones for HSP 60 by ELISA. Tissue samples from four patients with classic, active RA were used in this study. The isolated cells were electrofused in strongly hypo-osmolar medium with cells either of the mouse strain X63-Ag8-653 (Ag8) or the heteromyeloma strain HAB-1. Clones positive for IgG, the IgG fraction of the supernatant of the isolated synovial cells and the IgG of the serum of the patients were tested in an ELISA for reactivity to the recombinant HSP 60 or Yersinia enterocolitica, which shows great homology with mycobacterial HSP 65 and human HSP 60. The expression of this HSP 60 was studied in normal and rheumatoid synovial tissue using a polyclonal rabbit serum against HSP 60 from Y. enterocolitica (Ye HSP 60). In this way we investigate differences in the expression of HSP 60 and compared the pattern of this HSP60 with the pattern of mycobacterial HSP65 and human HSP 60 described by others. In three of four patients 10 IgG secreting B-cell clones showing a specificity for HSP 60 were detected. IgG specific for HSP 60 was also detected in the supernatant of the isolated synovial cells before fusion and in the serum of these patients. HSP 60 was demonstrated immunohistochemically within the rheumatoid synovial tissue and showed stronger expression with a different distribution when compared with the expression in normal synovial tissue. B-cell clones from rheumatoid synovial tissue thus exhibit a specificity for bacterial HSP 60, and a monospecific rabbit serum against this HSP shows strong reactivity within the rheumatoid synovial tissue. It may be postulated that a humoral HSP 60 response, initially directed against an infectious agent, could react with cross-reactive epitopes of rheumatoid synovial tissue or with self-HSP perpetuating the local inflammatory process.
多项研究表明，对热休克蛋白（HSP）具有特异性的T淋巴细胞在类风湿关节炎（RA）的发病机制中起作用。令人惊讶的是，目前尚无针对HSP具有特异性的B淋巴细胞的实验数据。为了研究类风湿滑膜组织中的B淋巴细胞是否对HSP 60具有特异性，我们通过电融合技术使滑膜组织B淋巴细胞永生化，并通过ELISA检测B细胞克隆对HSP 60的特异性。本研究使用了4例典型活动性RA患者的组织样本。将分离的细胞在强低渗培养基中与小鼠X63-Ag8-653（Ag8）品系细胞或异源骨髓瘤HAB-1品系细胞进行电融合。对分离的滑膜细胞上清液中的IgG组分、患者血清中的IgG以及IgG阳性克隆进行ELISA检测，以检测其对重组HSP 60或小肠结肠炎耶尔森菌的反应性，小肠结肠炎耶尔森菌与分枝杆菌HSP 65和人HSP 60具有高度同源性。使用针对小肠结肠炎耶尔森菌HSP 60（Ye HSP 60）的多克隆兔血清，研究正常和类风湿滑膜组织中该HSP 60的表达。通过这种方式，我们研究了HSP 60表达的差异，并将该HSP60的模式与其他人描述的分枝杆菌HSP65和人HSP 60的模式进行了比较。在4例患者中的3例中，检测到10个对HSP 60具有特异性的分泌IgG的B细胞克隆。在融合前分离的滑膜细胞上清液以及这些患者的血清中也检测到了对HSP 60具有特异性的IgG。在类风湿滑膜组织中通过免疫组织化学证实了HSP 60的存在，与正常滑膜组织中的表达相比，其表达更强且分布不同。因此，类风湿滑膜组织中的B细胞克隆对细菌HSP 60具有特异性，针对该HSP的单特异性兔血清在类风湿滑膜组织中显示出强烈反应性。可以推测，最初针对感染因子的体液HSP 60反应可能与类风湿滑膜组织的交叉反应性表位或自身HSP发生反应，从而使局部炎症过程持续存在。
