Shimomura T, Fujimura K, Takafuta T, Fujii T, Katsutani S, Noda M, Fujimoto T, Kuramoto A
Department of Haematology and Oncology, Hiroshima University, Japan.
Br J Haematol. 1996 Dec;95(4):732-7. doi: 10.1046/j.1365-2141.1996.d01-1967.x.
To determine whether clonal T cells accumulate in idiopathic thrombocytopenic purpura (ITP), we performed single-strand conformation polymorphism (SSCP) analysis to detect T-cell receptor (TCR) beta-chain usage of peripheral T cells. We detected significantly more oligoclonal T cells (15.5 +/- 8.9 bands representative for clonal T-cell expansions) in peripheral blood from ITP patients than from healthy donors (2.8 +/- 2.6 bands). Frequently used V beta genes in these accumulated T cells in ITP were V beta 3, 6, 10, 13.1 and 14. To determine whether these bands were derived from clonal T cells, presumably in a preactivated state, we established some T-cell clones (expressing CD4 and TCR V beta 6. 13.1. or 14) by nonspecific stimulation from patients peripheral mononuclear cells, and examined their clonotypes. Clonal identities for three out of seven clones tested were confirmed using SSCP analyses to compare the migration of their beta-chain complementarity determining region 3 (CDR3) cDNAs, expanded by polymerase chain reaction (PCR) with those from peripheral blood. Therefore, distinctive T-cell clones accumulated in the periphery in ITP and they may be related to the autoimmune-mediated destruction of platelets.
为了确定克隆性T细胞是否在特发性血小板减少性紫癜(ITP)中积聚,我们进行了单链构象多态性(SSCP)分析,以检测外周血T细胞的T细胞受体(TCR)β链使用情况。我们发现,与健康供者外周血(2.8±2.6条带)相比,ITP患者外周血中寡克隆T细胞(代表克隆性T细胞扩增的15.5±8.9条带)明显更多。ITP中这些积聚的T细胞中常用的Vβ基因是Vβ3、6、10、13.1和14。为了确定这些条带是否源自可能处于预激活状态的克隆性T细胞,我们通过对患者外周血单个核细胞进行非特异性刺激,建立了一些T细胞克隆(表达CD4和TCR Vβ6、13.1或14),并检测了它们的克隆型。通过SSCP分析比较7个测试克隆中3个克隆的β链互补决定区3(CDR3)cDNA(通过聚合酶链反应(PCR)扩增)与外周血来源的CDR3 cDNA的迁移情况,确认了克隆身份。因此,ITP患者外周血中积聚了独特的T细胞克隆,它们可能与自身免疫介导的血小板破坏有关。