Analysis of pathogenesis and establishment of novel diagnostic strategy of oral mucosal diseases by T cell receptorbased analysis

基于T细胞受体分析的口腔粘膜疾病发病机制分析及新诊断策略的建立

• 批准号: 13557179
• 负责人: NAKAMURA Seiji
• 金额: $ 8.7万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13557179/
• 关键词: Immunology; Oral mucosal disease; T cell receptor; Antigen; Cytokine; Diagnosis; Oral lichen planus; Sjogren's syndrome; シェーグレン症侯群; 口腔扁平上皮癌; 遺伝子クローニング; T細胞; 臨床検査

This research was addressed to identify a disease-specific T cell receptor (TCR) in oral mucosal diseases and to establish a diagnostic strategy by examining the presence of disease-specific T cells expressing the TCR in patients with the diseases. Results obtained in this research were as follows :1)Identification of disease-specific TCR genes : PCR-based analysis revealed that restricted Vβ gene families are frequently used in tumor-infiltrating lymphocytes of patients with oral squamous cell carcinoma (SCC). Further SSCP-based analysis found oligoclonal expansion of T cell clonotypes in some Vβ gene families. DNA sequencing of the unique TCR genes and establishment of T cell clones expressing the unique TCR are being performed.2)Induction of T cells recognizing tumor-associated antigenic peptides from peripheral blood of patients with oral SCC : Peripheral blood mononuclear cells from patients with oral SCC were cultured in vitro with 13 kinds of tumor-associated antigenic peptides, … More and were then examine their peptide-specific cytotoxic T lymphocyte (CTL) activities. As a result, peptide-specific CTL activities were most frequently induced with SART-1_<690>, SARI-2_<93>, and ART4_<75>. CTL activities against other peptides also tended to be inducible from patients in whom CTL activities against SART-1_<690>, SARI-2_<93>, and ART4_<75> were induced. In contrast, in 40% of the patients examined, no CTL activity was induced with any peptides.3)Expression of immune-regulating molecules in the lesions of patients with oral SCC : Expression of, various immune-regulating molecules in the lesions was immunohistochemically examined. As a result, in patients in whom CTL activities against tumor-associated antigenic peptides, an aberrant expression of CD80 and ICAM-1 on SCC cells was observed. In contrast, an aberrant expression of tumor-associated antigen RCASI on SCC cells was observed in patients in whom no CTL activity was induced with any peptides. Apoptotic T cells were frequently observed around SCC cells expressing RCAS1, suggesting RCASI plays an important role in the tumor escape mechanism from the host immune system.This research indicated that tumor-associated antigenic peptides including SARI-1_<690>, SART-2_<93>, and ART4_<75> are useful in the diagnosis and follow-up of patients with oral SCC, and also for the establishment of cancer immunotherapy by peptide vaccination. However, it was also indicated that the antigenecity of tumor cells and the tumor escape mechanism from the host immune system must be paid attention. Novel diagnostic strategy to quantify the presence of disease-specific T cells in the lesions or peripheral blood from patients with various oral mucosal diseases is being also established. Less

本研究旨在鉴定口腔粘膜疾病中的疾病特异性T细胞受体（TCR），并通过检查疾病患者中表达TCR的疾病特异性T细胞的存在来建立诊断策略。1）疾病特异性TCR基因的鉴定：基于PCR的分析显示，限制性Vβ基因家族在口腔鳞状细胞癌（SCC）患者的肿瘤浸润淋巴细胞中频繁使用。进一步基于SSCP的分析发现在某些Vβ基因家族中T细胞克隆型的寡克隆扩增。正在进行独特TCR基因的DNA测序和表达独特TCR的T细胞克隆的建立。2）从口腔SCC患者外周血中诱导识别肿瘤相关抗原肽的T细胞：将来自口腔SCC患者的外周血单个核细胞与13种肿瘤相关抗原肽一起体外培养， ...更多信息 然后检测其肽特异性细胞毒性T淋巴细胞（CTL）活性。结果，SART-1_<690>、SARI-2_<93>和ART 4_最常诱导肽特异性CTL活性<75>。针对其他肽的CTL活性也倾向于从其中针对SART-1_<690>、SARI-2_<93>和ART 4_<75>的CTL活性被诱导的患者中被诱导。相反，在40%的受检患者中，任何肽均未诱导CTL活性。3）口腔SCC患者病变中免疫调节分子的表达：免疫化学检测病变中各种免疫调节分子的表达。结果，在对肿瘤相关抗原肽具有CTL活性的患者中，观察到SCC细胞上CD 80和ICAM-1的异常表达。相反，在没有用任何肽诱导CTL活性的患者中观察到SCC细胞上肿瘤相关抗原RCASI的异常表达。本研究表明，SARI-1_<690>、SART-2_<93>和ART 4_<75>等肿瘤相关抗原肽在口腔鳞癌的诊断和随访中具有重要意义，并可用于通过肽疫苗建立肿瘤免疫治疗。但同时也提示，肿瘤细胞的抗原性和肿瘤逃避宿主免疫系统的机制必须引起重视。新的诊断策略，以量化存在的疾病特异性T细胞的病变或外周血中的各种口腔粘膜疾病的患者也正在建立。少

项目成果

T cell receptor Vβgene usage by T cells reactive with the tumor-rejection antigen SARI-1 in oral squamous cell carcinoma.

口腔鳞状细胞癌中 T 细胞与肿瘤排斥抗原 SARI-1 反应的 T 细胞受体 Vβ 基因的使用。

• 发表时间: 2004
• 期刊: International Journal of Cancer 108
• 作者: Kumamaru;W.;et al.
• 通讯作者: et al.

Accumulation of oligoclonal T cells in the infiltrating lymphocytes in oral lichen planus

• DOI: 10.1034/j.1600-0714.2003.00143.x
• 发表时间: 2003-05-01
• 期刊: JOURNAL OF ORAL PATHOLOGY & MEDICINE
• 影响因子: 3.3
• 作者: Kawamura, E;Nakamura, S;Shirasuna, K
• 通讯作者: Shirasuna, K

T cell receptor Vβ gene usage by T cells reactive with the tumor-rejection antigen SARI-1 in oral squamous cell carcinoma.

口腔鳞状细胞癌中与肿瘤排斥抗原 SARI-1 发生反应的 T 细胞对 T 细胞受体 Vβ 基因的使用。

• 发表时间: 2004
• 期刊: International Journal of Cancer 108
• 作者: Kumamaru;W.;Nakamura;S.;Kadena;T.;Yamada;A.;Kawamura;E.;Sasaki;M.;Ohyama;Y.;Toyoshima;T.;Hayashida;J.;Itoh;K.;Shirasuna;K.
• 通讯作者: K.

Thunawaki, S., et al.: "Possible function of salivary gland epithelial cells as nonprofessional antigen-presenting cells in the development of Sjogren's syndrome"Journal of Rheumatology. 29. 1041-1046 (2002)

Thunawaki, S. 等人：“唾液腺上皮细胞作为非专业抗原呈递细胞在干燥综合征发展中的可能功能”风湿病杂志。

Kawamura, E., et al.: "Accumulation of oligoclonal T cells in the infiltrating lymphocytes in oral lichen planus"Journal of Oral Pathology & Medicine. (in press). (2003)

Kawamura, E., et al.：“口腔扁平苔藓浸润淋巴细胞中寡克隆 T 细胞的积累”口腔病理学杂志