Preparation of human monoclonal antibodies using human B cell producing autoantibodies and analyses of novel autoantigens.

使用产生自身抗体的人 B 细胞制备人单克隆抗体并分析新型自身抗原。

• 批准号: 60480204
• 负责人: YACHI Akira
• 金额: $ 3.65万
• 依托单位: Sapporo Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60480204/
• 关键词: Autoantibody; Autoantigen; Human monoclonal antibody; エピトープ

In some cases with autoimmune diseases and malignancies, autoantibodies to unknown antigens were detected in the sera. Using monoclonal antibodies, we have studied these autoantigens and autoantibodies and analysed the immune system. In order to prepare human type monoclonal antibodies, human B cells from cases with autoimmune diseases and malignancies were used. Therein we experienced a colonic cancer patient with anti-CEA monoclonal antibody. For parent cells, human myeloma cell line ARH77, human plasma cell line UC729-6 and mouse myeloma cell line P3-X63-Ag8-653 were served. Human cell lines which we used for fusion gave little hybridomas, but many human monoclonal antibodies were obtained when P3-X63-Ag8-653 was used. By a fusion with B cells in pleural effusion of a gastric cancer patient, human monoclonal antibody 3B7 (IgG) was raised, showing a staining reactivity with gastric and colonic cancers but not with non cancerous tissues. Molecular profile of autoantigen to which monoclonal antibody 3B7 reacts is under a further investigation. At present we are trying to raise hybridomas from new parent cells KR-4 and KR-12 using a newly developed electrofusion method.

在一些自身免疫性疾病和恶性肿瘤的病例中，血清中检测到针对未知抗原的自身抗体。利用单克隆抗体，我们研究了这些自身抗原和自身抗体，并分析了免疫系统。为了制备人型单克隆抗体，使用了来自自身免疫性疾病和恶性肿瘤病例的人B细胞。其中我们遇到了一个结肠癌患者抗cea单克隆抗体。亲本细胞为人骨髓瘤细胞系ARH77、人浆细胞系UC729-6和小鼠骨髓瘤细胞系P3-X63-Ag8-653。我们用人细胞株进行融合，得到的杂交瘤很少，而用P3-X63-Ag8-653进行融合，得到的人单克隆抗体较多。通过与胃癌患者胸腔积液中的B细胞融合，产生人单克隆抗体3B7 (IgG)，对胃癌和结肠癌有染色反应，但对非癌组织无反应。单克隆抗体3B7与自身抗原反应的分子谱还有待进一步研究。目前我们正在尝试用一种新开发的电融合方法从新的亲本细胞KR-4和KR-12中培养杂交瘤。

项目成果

Imai,K., Yachi,A., et al.: "Adenocarcinoma-associated YH206 antigen in the sera of cancer patients." Labeled Antibodies ( Kano,K. ed., Tokyo University Press, Tokyo ). 433-436 (1987)

Imai,K.、Yachi,A. 等人：“癌症患者血清中的腺癌相关 YH206 抗原。”

Sugiyama,T.,Yachi,A.,et al: Jpn.J.Cancer Res(Gann). 77. 722-729 (1986)

Sugiyama,T.、Yachi,A. 等人：Jpn.J.Cancer Res（江恩）。

Yachi,A., et al.: "Immunohistological distribution of antigenic determinants YH206 detected by a monoclonal antibody." Jpn. J. Med.25. 127-134 (1986)

Yachi,A., et al.：“通过单克隆抗体检测到的抗原决定簇 YH206 的免疫组织学分布。”

Sugiyama,T., Yachi,A., et al.: "Preparation of human monoclonal antibodies of IgG type to gastro-intestinal cancer-associated antigen." Jpn. J. Cancer Res. (gann). 77. 722-729 (1986)

Sugiyama,T., Yachi,A., et al.：“针对胃肠癌相关抗原的 IgG 型人单克隆抗体的制备”。

Imai,K.,Yachi,A.,et al Kano,K.ed.: "Labeled Antibodies"Tokyo University Press. 433-436 (1987)

Imai,K.、Yachi,A.、et al Kano,K.ed.：“标记抗体”东京大学出版社。