Making the antibody library from pathologic lesion of intestinal bowel disease, and analysis and application of antibody and antigen.

我们的研究目的是寻找有用的抗体和抗原来分析炎症性肠病（IBDs）的发病机制和诊断和治疗。在IBDs的病变中，大量产生抗体的浆细胞浸润。然而，这些免疫细胞中产生的抗体所识别的抗原仍不清楚。理论上，病灶内局部分泌的抗体必然与疾病过程密切相关。因此，我们计划进行一项研究，直接分析浸润在病变中的浆细胞产生的抗体。在第一年，我们检测了小鼠炎症性结肠病变中cDNA编码抗体的克隆性，这些抗体是由葡聚糖硫酸钠引起的，葡聚糖硫酸钠代表了小鼠溃疡性结肠炎模型。我们确定了在结肠病变内浸润的抗体产生细胞的克隆性。因此，我们认为cDNA编码抗体的克隆性可能是鉴定病变特异性抗体的重要依据。第二年，我们分析了辣根过氧化物酶（HRP）免疫大鼠区域淋巴结中抗体cDNA的克隆性。利用结组织cDNA合成单链Fv （scFv）单克隆抗体，研究其HRP反应性。我们证明了结组织cDNA编码重链和轻链的克隆性。然而，在ELISA系统中，由cDNA随机合成的单克隆抗体缺乏与HRP的结合。为了在淋巴结中鉴定编码抗hrp抗体的cDNA，我们筛选了免疫淋巴结组织的cDNA文库。只观察到一种类型的抗体克隆。经随机序列分析，该抗hrp单克隆抗体的重链cDNA与多个克隆中的一个相对应。然而，抗hrp抗体的轻链cDNA序列与序列完全一致，没有克隆性。这些结果表明，我们遗憾的是，cDNA克隆编码的抗体分子并不一定表明病变特异性抗体。为了克服这一困难，目前正在进行进一步的分析。少

The purpose of our research is to find useful antibodies and antigens for analyzing the pathogenesis and for diagnosing and treating inflammatory bowel diseases (IBDs). In the lesions of IBDs, numbers of antibody-producing plasma cells infiltrate. However, the antigens recognized by antibodies produced in these immune cells remain unclear. Theoretically, the antibodies locally secreted within the lesions must be closely correlated with the disease process. We thus planned a research directly analyzing the antibodies produced by the plasma cells infiltrating in the lesion.In the first year, we examined the clonality of cDNA encoding antibodies in mouse inflammatory colonic lesions experimentally provoked by dextran sodium sulfate administration, which represents a mouse model of ulcerative colitis. We identified clonalities of the antibody-producing cells infiltrating within the colonic lesion. Therefore, we assumed that the clonality of cDNA encoding antibodies might be useful as crite … More ria for identifying lesion-specific antibodies.In the second year, we analyzed the clonality of antibody cDNA in regional lymph nodes obtained from horseradish peroxidase (HRP)-immunized rats. The HRP reactivity of single chain Fv (scFv) monoclonal antibodies synthesized from nodal tissue cDNA was pursued. We demonstrated clonalities in the nodal tissue cDNA encoding both the heavy and light chains. However, scFv monoclonal antibodies randomly synthesized from the cDNA lacked binding with HRP in the ELISA system. In order to identify cDNA encoding anti-HRP antibodies in the lymph nodes, we screened the cDNA library made from the immunized nodal tissue. Only one type of antibody clone was observed. By random sequence analysis, the heavy chain cDNA of this anti-HRP monoclonal antibody corresponded to one of the several clones. However, the light chain cDNA of the anti-HRP antibody was identical to the sequence that did not show any clonality. These results suggest to our regret that cDNA clones encoding antibody molecules do not necessarily indicate the lesion-specific antibodies. To overcome this difficulty, further analysis is now continuing. Less