Fenning S, Wolff-Vorbeck G, Hackl W, Krawinkel U, Lührmann R, Northemann W, Peter H H, Schlesier M
Abteilung Rheumatologie und Klinische Immunologie, Chirurgische Universitätsklinik, Freiburg, Germany.
Clin Exp Immunol. 1995 Sep;101(3):408-13. doi: 10.1111/j.1365-2249.1995.tb03127.x.
In sera of patients with mixed connective tissue disease (MCTD) high titres of IgG autoantibodies to U1snRNP-specific proteins (70 kD, A, C) are found, suggesting an antigen-driven and T-cell-dependent process. In order to establish U1snRNP-specific T cell lines we cultured under various culture conditions mononuclear cells from MCTD patients and healthy donors with a highly purified UsnRNP preparation from HeLa cells. Nine T cell lines were established by limiting dilution cloning from two MCTD patients and five T cell lines from a healthy individual. All T cell lines expressed the TCR alpha beta/CD3 complex. Surprisingly, most of the T cells lines exhibited the CD8 phenotype. Irrespective of this phenotype, all T cell lines showed a proliferative response to an N-terminal part (aa 51-195) of recombinant U1-specific 70-kD protein. One CD8+ T cell clone exhibited cytotoxic activity against an autologous B cell line pulsed with snRNP or recombinant fragments (aa 51-95 and aa 51-88). Interestingly, two T cell lines proliferated in response to four recombinant polypeptides representing different parts of the U1snRNP 70-kD protein. Since regions of sequence homology are distributed over the 70-kD molecule, it is suggested that conserved motifs may be recognized by the T cell lines.
在混合性结缔组织病(MCTD)患者的血清中,发现了针对U1snRNP特异性蛋白(70 kD、A、C)的高滴度IgG自身抗体,提示这是一个抗原驱动且依赖T细胞的过程。为了建立U1snRNP特异性T细胞系,我们在多种培养条件下,用来自HeLa细胞的高度纯化的U1snRNP制剂培养了MCTD患者和健康供体的单核细胞。通过有限稀释克隆从两名MCTD患者中建立了9个T细胞系,从一名健康个体中建立了5个T细胞系。所有T细胞系均表达TCRαβ/CD3复合物。令人惊讶的是,大多数T细胞系表现出CD8表型。无论这种表型如何,所有T细胞系对重组U1特异性70-kD蛋白的N端部分(氨基酸51-195)均表现出增殖反应。一个CD8+T细胞克隆对用snRNP或重组片段(氨基酸51-95和氨基酸51-88)脉冲处理的自体B细胞系具有细胞毒性活性。有趣的是,两个T细胞系对代表U1snRNP 70-kD蛋白不同部分的四种重组多肽有增殖反应。由于序列同源区域分布在70-kD分子上,提示保守基序可能被T细胞系识别。