SEREX ANALYSIS OF IPF PATIENS : DEMONSTRATION OF SOME AUTOANTIGENS REACTED WITH CD4 POSITIVE T CELLS IN BRONCHOALVEOLAR LAVAGE FLUIDS

特发性肺纤维化（IPF）的自然史总是逐渐和进行性恶化，从诊断时起的中位生存期为2.5-3.5年，但仍不确定导致这种临床恶化的原因。为了鉴定可能在PF背景下起作用的抗原，使用cDNA表达文库的血清学分析（SEREX），使用14名IPF患者的血清筛选II型肺泡细胞癌（A549）细胞系的cDNA噬菌体文库。我们的实验包括分析支气管肺泡灌洗液（BALF）中的TCR-Vb库和CD 4阳性T细胞的寡克隆性。我们的研究结果表明，在一些IPF患者的BALF中的CD 4阳性T细胞寡克隆扩增，这表明通过TCR识别主要组织相容性复合物上的限制性表位来进行抗原驱动的刺激。例2中，BALF标本中两个TCR-Vb重排（TCR-Vb 2和Vb 7）在DGGE上与VATS活检（2个月后）中相同。这表明BALF中的一些浸润性CD 4阳性T细胞共享自身抗原上的表位。在12例IPF患者中的4 - 5例中检测到膜联蛋白I、磷酸甘油酸激酶1、膜联蛋白IV、bax抑制剂1和细胞色素c氧化酶亚基Va的自身抗体。病例2中BALF和VATS样品中相同TCR-Vb 7克隆的CDR 3区与膜联蛋白1的部分（残基20-26）具有高度同源性。该区域包括在通过用DRB 10410和DRB 11307（HLA-DR）的TEPITOPE分析选择的膜联蛋白1的表位中

The natural history of idiopathic lung fibrosis(IPF) is invariably one of gradual and progressive deterioration, with median length of survival from the time of diagnosis ranging 2.5-3.5 yrs, it still remains uncertain what causes such clinical deterioration. To identify antigens that may play a role in the context of PF, a cDNA phage library of type II alveolar cell carcinoma (A549) cell lines were screened using sera of 14 patients with IPF using serological analysis of the cDNA expression library(SEREX). Our experiments included an analysis of TCR-Vb repertoire and oligoclonality of CD4 positive T cells in bronchoalveolar lavage fluid(BALF). Our results showed that CD4 positive T cells in BALF of some patients with IPF expand oligoclonally, suggesting antigen-driven stimulation by recognizing a restricted epitope on the major histocompatibility complex through TCR. In cases 2, two TCR-Vb rearrangements (TCR-Vb2 and Vb7) in BAL sample on DGGE were identical to those in the VATS biopsy (2 months later). This suggests that some infiltrating CD4 positive T cells in BALF shared epitopes on autoantigens. Autoantibodies to annexin I, phosphoglyverate kinase 1, annexin IV, bax inhibitor 1, and cytochrome c oxidase subunit Va was detected in 4 to 5 of 12 patients with IPF. The CDR3 region of identical TCR-Vb7 clones in both BALF and VATS samples in case 2 have high homology with the portion (residues 20-26) of annexin 1. This region was included in epitopes of annexin 1 selected by TEPITOPE analysis with DRB1 0410 and DRB1 1307 (HLA-DR