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Identification of T cell epitopes and development of a new immunotherapy for idiopathic thrombocytopenic purpura

T细胞表位的鉴定和特发性血小板减少性紫癜新免疫疗法的开发

基本信息

批准号：
16590942
负责人：
HATO Takaaki
金额：
$ 2.24万
依托单位：
Ehime University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590942/
关键词：
platelet ITP GPIIb-IIIa T cell autoimmunity integrin epitope

项目摘要

The purpose of this study is to define the role and dominant epitopes of GPIIb-reactive T cells in idiopathic thrombocytopenic purpura (ITP). We established a CD4^+ T cell line recognizing the GPIIb #429-443 amino acid sequence in a HLA-DR^*0405-restricted manner from an ITP patient. Interferon-gamma production was detected when the cells were co-cultured autologous immature dendritic cells which had been loaded with platelet lysate, suggesting that the region around GPIIb 429-443 can be presented to CD4^+ T cells in the context of HLA-DRB1^*0405. We isolated further T cell clones from several ITP patients. These clones were mainly CD4^+ helper T cells, but CD8^+ cytotoxic T cells could also be isolated. To measure helper activity of T cells, we established a sandwich ELISA assay for GPIIb antibody using anti-GPIIb-IIIa monoclonal antibodies produced in our laboratory. Then we reported WT1-specific and HLA class II-restricted CD4^+ T cells possessing direct cytotoxic activity against leukemia cells, indicating establishment of specific assay for cytotoxic activity of CD4^+ T cells. We also reported that perforin is expressed constitutively in memory CD8^+ cytotoxic T cells, but is dependent on cell activation in memory CD4^+ cytotoxic T cells, suggesting differential regulation of cytotoxicity in CD4^+ and CD8^+ T cells. Then we attempted to identify the amino acid regions critical for GPIIb-IIIa activation using alanine-scaninng mutagenesis. We identified three amino acids critical for ligand binding to GPIIb-IIIa and an amino acid motif in the intracellular domain of GPIIb that suppresses GPIIb-IIIa activation.

本研究的目的是确定GPIIb反应性T细胞在特发性血小板减少性紫癜（ITP）中的作用和优势表位。我们建立了一个以HLA-DR^*0405限制性方式识别ITP患者GPIIb #429-443氨基酸序列的CD 4 ^+ T细胞系。当这些细胞与自体未成熟树突状细胞共培养时，检测到干扰素-γ的产生，这些树突状细胞已经加载了血小板裂解物，这表明GPIIb 429-443周围的区域可以在HLA-DRB 1 ^*0405的背景下呈递给CD 4 ^+ T细胞。我们从几个ITP患者中分离出进一步的T细胞克隆。这些克隆主要是CD 4 ^+辅助性T细胞，但也可以分离出CD 8 ^+细胞毒性T细胞。为了测量T细胞的辅助活性，我们使用本实验室制备的抗GPIIb-IIIa单克隆抗体建立了GPIIb抗体的夹心ELISA测定法。随后，我们报道了WT 1特异性和HLA II类限制性CD 4 ^+ T细胞对白血病细胞具有直接的细胞毒活性，从而建立了特异性的CD 4 ^+ T细胞毒活性检测方法。我们还报道了穿孔素在记忆性CD 8 ^+细胞毒性T细胞中组成型表达，但在记忆性CD 4 ^+细胞毒性T细胞中依赖于细胞活化，这表明CD 4 ^+和CD 8 ^+ T细胞的细胞毒性调节存在差异。然后，我们尝试使用丙氨酸扫描诱变来鉴定GPIIb-IIIa激活的关键氨基酸区域。我们确定了三个氨基酸的关键配体结合GPIIb-IIIa和一个氨基酸基序在细胞内域的GPIIb抑制GPIIb-IIIa激活。

项目成果

期刊论文数量（46）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Critical residues for ligand binding in blade 2 of the propeller domain of the integrin αIIb subunit.

整合素 αIIb 亚基螺旋桨结构域叶片 2 中配体结合的关键残基。

DOI：
发表时间：
2004
期刊：
Thrombosis and Haemostasis 91・1
影响因子：
0
作者：
Niiya H;Sakai I;Lei J;Azuma T;Uchida N;Yakushijin Y;Hato T;Fujita S;Yasukawa M.;Yang Y;Tamura T et al.
通讯作者：
Tamura T et al.

Identification of an epitope on glycoprotein IIb-IIIa that is recognized by HLA-DRB1^*0405-restricted CD4^+ T cells from a patient with immune thrombocytopenic purpura.

鉴定糖蛋白 IIb-IIIa 上的表位，该表位可被免疫性血小板减少性紫癜患者的 HLA-DRB1^*0405 限制性 CD4^ T 细胞识别。

DOI：
发表时间：
2004
期刊：
Journal of Thrombosis and Haemostasis 2(2)
影响因子：
0
作者：
Yamanouchi J;Hato T;Tamura T;Fujita S;Yasukawa M.
通讯作者：
Yasukawa M.

Identification of an epitope on glycoprotein IIb-IIIa that is recognized by HLA-DRB1*0405-restricted CD4^+ T cells from a patient with immunne thrombocytopenic purpura.

糖蛋白 IIb-IIIa 上表位的鉴定，该表位被免疫性血小板减少性紫癜患者的 HLA-DRB1*0405 限制性 CD4+ T 细胞识别。

DOI：
发表时间：
2004
期刊：
Journal of Thrombosis and Haemostasis 2・2
影响因子：
0
作者：
Niiya H;Sakai I;Lei J;Azuma T;Uchida N;Yakushijin Y;Hato T;Fujita S;Yasukawa M.;Yang Y;Tamura T et al.;Yamamoto K. 他;Takeuchi s;Yamanouchi J et al.;Matsushita C;Yamanouchi et al.
通讯作者：
Yamanouchi et al.

Existence of leukemic clones resistant to both imatinib mesylate and rituximab before drug therapies in a patient with Philadelphia chromosome-positive acute lymphocytic leukemia