Shimizudani N, Murata H, Keino H, Kojo S, Nakamura H, Morishima Y, Sakamoto T, Ohtsuka M, Sekisawa K, Sumida M, Sumida T, Matsuoka T
Department of Internal Medicine, University of Tsukuba, Japan.
Clin Exp Immunol. 2002 Jul;129(1):140-9. doi: 10.1046/j.1365-2249.2002.01871.x.
Idiopathic pulmonary fibrosis (IPF) is an inflammatory lung disease characterized by the accumulation of inflammatory cells and deposition of collagen, resulting in lung remodelling. High numbers of T cells are present in bronchoalveolar lavage fluid (BALF) of IPF patients, although the characteristics of these cells are yet to be determined. To elucidate the pathogenic mechanisms of IPF, we analysed the T cell receptor (TCR) of BALF lymphocytes in three patients with IPF and three healthy subjects as control. TCR repertoire of BALF lymphocytes and T cell clonality were examined by family PCR and Southern blot analysis, and single-strand conformation polymorphism (SSCP), respectively. We observed that the TCR repertoire in the lung was heterogeneous, both in the control subjects and three patients with IPF. SSCP analysis demonstrated an increase in the number of accumulated T cell clones in BALF of two of the three patients, but not in the healthy subject. Furthermore, junctional sequence analysis showed the presence of conserved amino acid motifs (ETGRSG, LAxG, QGQ, GxQP, GRxG, VAR, PGT, GTI, GGT, TGR, LxLxQ, SGQ) in the TCR-CDR 3 region of BAL lymphocytes in patients with IPF, whereas only two amino acid motifs (VTTG, GGE) were found in the control. Our findings suggest that T cells in BALF of patients with IPF expand oligoclonally in the lung, suggesting antigen stimulation of these cells.
特发性肺纤维化(IPF)是一种炎症性肺部疾病,其特征为炎症细胞积聚和胶原蛋白沉积,导致肺重塑。IPF患者的支气管肺泡灌洗液(BALF)中存在大量T细胞,尽管这些细胞的特征尚待确定。为了阐明IPF的致病机制,我们分析了3例IPF患者和3名健康对照者BALF淋巴细胞的T细胞受体(TCR)。分别通过家族聚合酶链反应(PCR)和Southern印迹分析以及单链构象多态性(SSCP)检测BALF淋巴细胞的TCR库和T细胞克隆性。我们观察到,无论是健康对照者还是3例IPF患者,肺中的TCR库都是异质性的。SSCP分析显示,3例患者中有2例的BALF中积累的T细胞克隆数量增加,而健康对照者中则没有。此外,连接序列分析显示,IPF患者BAL淋巴细胞的TCR - CDR 3区域存在保守的氨基酸基序(ETGRSG、LAxG、QGQ、GxQP、GRxG、VAR、PGT、GTI、GGT、TGR、LxLxQ、SGQ),而在对照组中仅发现两个氨基酸基序(VTTG、GGE)。我们的研究结果表明,IPF患者BALF中的T细胞在肺中呈寡克隆性扩增,提示这些细胞受到了抗原刺激。