Development of a laboratory examination system for T cell clonality analysis.

开发用于 T 细胞克隆分析的实验室检查系统。

• 批准号: 07557227
• 负责人: YAMAMOTO Kazuhiko
• 金额: $ 3.84万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557227/
• 关键词: T Cell Receptor; PCR; SSCP; T Cell; Rheumatoid Arthritis; Systemic Lupus Erythematosus

To investigate T cell involvement in antigen-specific immune responses, it is important to detect accumulating T cells at a clonal level in vivo.however, thus far the clinical application of sush analyzes has been limeted. Here we have established novel primers to anneal with T cell reseptor (TCR) beta genes of multiple Vbeta families and applied them to reverse transcription-polymerase chain reaction-single strand conformation polymorphism (RT-PCR-SSCP) analysis to evaluate peripheral T cell clonality of autoimmue disease patients. As a result, the new Vbeta primers could detect accumulating T cell clones in the periphery of healthy individuals and patients. It was revealed that patients with autoimmune diseases, such as systemic lupus erythematosus (SLE) , had a larger number of clonal accumulations of peripheral T cells compared with normal individuals.thus, the RT-PCR-SSCP systemic using the new multifamily Vbeta primers is the first such laboratory examination to detect T cell clonal expantion, and will provide a simple and sensitive tool to aid in the diagnosis and also in the investigation of the pathoqenes of autoimmune diseases.

为了研究T细胞在抗原特异性免疫反应中的作用，在活体水平上检测蓄积的T细胞是很重要的。然而，到目前为止，SUH分析的临床应用一直受到限制。在这里，我们建立了新的引物，与多个Vbeta家族的T细胞受体(TCR)β基因进行退火，并将其应用于逆转录-聚合酶链式反应-单链构象多态性(RT-PCR-SSCP)分析，以评估自身免疫性疾病患者的外周T细胞克隆性。因此，新的Vbeta引物可以检测到健康人和患者外周血中积累的T细胞克隆。研究发现，系统性红斑狼疮(SLE)等自身免疫性疾病患者外周血中T细胞的克隆性聚集较正常人多，因此，采用新型多家族Vbeta引物的RT-PCR-SSCP系统是首次检测T细胞克隆性扩增的实验室检测方法，将为自身免疫性疾病的诊断和发病机制的研究提供一种简便、灵敏的工具。

项目成果

Atsushi Nakajima, Tatsuhiko Kodama, Yoshio Yazaki, Masakazu Takazoe, Norifumi Saito, Ryoichi Suzuki, Haruo Nishino, Kazuhiko Yamamoto, Jack Silver, and Nobuyuki Matsuhashi: "Specific clonal T cell accumulation in intestinal lesions of crohn's disease" The

Atsushi Nakajima、Tatsuhiko Kodama、Yoshio Yazaki、Masakazu Takazoe、Norifumi Saito、Ryoichi Suzuki、Haruo Nishino、Kazuhiko Yamamoto、Jack Silver 和 Nobuyuki Matsuhashi：“克罗恩病肠道病变中的特异性克隆 T 细胞积聚”

Atsushi Tanaka: "Clonotypic analysis of T cells infiltrating the skin of patients with atopic Dermatitis:Evidence for antigen-driven accumulation of T cells" Human Immunology. 48. 107-113 (1996)

Atsushi Tanaka：“特应性皮炎患者皮肤浸润 T 细胞的克隆型分析：抗原驱动 T 细胞积累的证据”人类免疫学。

Tanaka,A.: "Clonotypic analysis of T cells infiltrating the skin of patients with atopic dermatitis:evidence for antigten-driven accum ulation of T cells." Hum.Immunol.48. 107-113 (1996)

Tanaka,A.：“特应性皮炎患者皮肤浸润的 T 细胞的克隆型分析：抗原驱动 T 细胞积累的证据。”

Yamamoto, K.: "Accumulation of distinct T cell clonotypes in human solid tumors" J. Immunol.154. 1804-1809 (1995)

Yamamoto, K.：“人类实体瘤中不同 T 细胞克隆型的积累”J.Immunol.154。

Kazuhiko Yamamoto: "Significance of antigen-specific T cell clones in collagen diseases : Analysis with a novel T cell clonality evaluation system" Internal Medicine. 36(4). 242-247 (1997)

Kazuhiko Yamamoto：“抗原特异性 T 细胞克隆在胶原蛋白疾病中的意义：使用新型 T 细胞克隆性评估系统进行分析”内科。