Analysis of Abnormal DNA Rearrangement of Ig Heavy Chain Gene Loci in Precursor B Cell Lines Derived From Primary Immunodeficiency Patients

原发性免疫缺陷患者来源的前体 B 细胞系中 Ig 重链基因座的异常 DNA 重排分析

• 批准号: 01570523
• 负责人: MATSUOKA Hiroshi
• 金额: $ 1.34万
• 依托单位: Nagoya University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01570523/
• 关键词: Primary immunodeficiency; Precursor B cell line; Abnormal DNA rearrangement; Recombinase; Severe combined immunodeficiency; X-Linked agammaglobulinemia; Common variable immunodeficiency; 伴性無ガンマグロブリン血症; Common variable immunodeficiency; 重症複合型免疫不全症

1). EB virus transformed (precursor) B cell lines (139 cell lines) were established from 8 Severe Combined immunodeficiency (SCID) patients, one X-linked agammaglobulinemia (XLA) patient and 8 common variable immunodeficiency (CVID) patients.2). Fourteen of 139 cell lines had precursor B cell phenotypes, shown by immunofluorescent technique.3). The study of DNA rearrangements of Ig heavy chain gene loci in these B cell lines showed a defect of DJ-VDJ rearrangement in one XLA and one CVID patient, and showed a defect of DJ rearrangement in SCID patient.4). RAG-1 mRNA was detected in SCID 86730 cell clone, which had DJ rearrangement defect suggesting of the abnormal recombinase system, by using RT-PCR method.5). Recombinant vectors consisted of signal sequences, 12/23 spacer, Neo gene (inverted), HygroB gene and SRalpha promoter were constructed and used to measure recombinase activities of these B cell lines.

1）。 EB 病毒转化（前体）B 细胞系（139 个细胞系）是从 8 名严重联合免疫缺陷 (SCID) 患者、1 名 X 连锁无丙种球蛋白血症 (XLA) 患者和 8 名常见变异型免疫缺陷 (CVID) 患者建立的。2)。免疫荧光技术显示，139 个细胞系中有 14 个具有前体 B 细胞表型。3)。对这些B细胞系中Ig重链基因位点DNA重排的研究显示，一名XLA和一名CVID患者存在DJ-VDJ重排缺陷，并且SCID患者存在DJ重排缺陷。4)。 RT-PCR方法在SCID 86730细胞克隆中检测到RAG-1 mRNA，该细胞克隆存在DJ重排缺陷，提示重组酶系统异常。5)。构建了由信号序列、12/23间隔区、Neo基因（反向）、HygroB基因和SRα启动子组成的重组载体，并用于测量这些B细胞系的重组酶活性。

项目成果

柘植 郁哉: "病因への分子生物学的アプロ-チー無ガンマグロブリン血症" 臨床免疫. 22. 791-800 (1990)

Ikuya Tsuge：“无丙种球蛋白血症发病机制的分子生物学方法”临床免疫学 22. 791-800 (1990)。

H, Matsuoka: "Severe combined immunodeficiency" Jpn J. Pediatric Medicine. 20. 700-705 (1990)

H，Matsuoka：“严重联合免疫缺陷”Jpn J.儿科医学。

Y.Kamachi: "RAGー1 message detected in precursor B cell lines derived from severe combined immunodeficiency patients"

Y.Kamachi：“在源自严重联合免疫缺陷患者的前体 B 细胞系中检测到 RAGー1 信息”

松岡 宏: "重症複合型免疫不全症" 小児内科. 20. 700-705 (1990)

Hiroshi Matsuoka：“严重联合免疫缺陷”小儿内科学 20. 700-705 (1990)。

I. Tsuge: "Molecularbiological Approach for Pathogenesis-Agammaglobulinemia" Clinical Immunology. 22. 791-800 (1990)

I. Tsuge：“发病机制的分子生物学方法-无丙种球蛋白血症”临床免疫学。