Using soluble HLA-peptide tetramer, the studies on the cloning and immune-response monitoring of melanoma antigen-specific cytotoxic lymphocyte
利用可溶性HLA肽四聚体进行黑色素瘤抗原特异性细胞毒性淋巴细胞克隆及免疫反应监测的研究
基本信息
- 批准号:13670901
- 负责人:
- 金额:$ 2.3万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2001
- 资助国家:日本
- 起止时间:2001 至 2002
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
The purpose of this project is the studies on the cloning and immune-response monitoring of the melanoma antigen-specific cytotoxic T lymphocytes using soluble peptide-HLA tetramers produced.1. The tumor-infiltrating T-lymphocyte (TIL) 620 line (recognized gp-100-209) was double-stained with anti-CD8 mAb-PC5 and HLA-A2.1/gp100-209 PE-tetramer. The tetramer-binding CD8+ T cells (14 %) were sorting from TIL 620 line by fluorescence-activated cell sorting (FACS) and expanded in vitro. The cultured tetramer-binding CD8+ T cells were stained 98.4 % with HLA-A2.1/gp 100-209 PE-tetramer and released IFN-γ more than 7 fold of that from original TIL 620 line. It become is able to cloning the antigen-specific active T lymphocytes using HLA tetramers.2. HLA-A2.6 and A2.7 heavy chains were constructed from HLA-A2.1 by mutagenesis kit, synthesized and purified using a prokaryotic expression system. HLA-A2.6/gp100-280 tetramer was produced, and stained 4-5 % of TIL 660 (recognized A2.l/gp-100-280 epitope) and 7.97 % of the induced T cell line from PBMC of HLA-A2.6+ healthy donor by gp100-280 peptide. These results suggested that the HLA-A2.1 restricted melanoma-antigen peptide might be cross-recognized by other A2-supertype molecules.3. As the model of monitoring tumor-specific T lymphocytes using HLA tetramers, fluorescent HLA-CMV peptide tetramer were used to monitor the recovery of CMV-specific T lymphocytes in recipients of allogeneic stem cell transplants, (1) CMV pp65-495 specific T cells were induced, using CMV pp65-495 peptide, in 7 of 8 PMBC from A2.1+ healthy donor, and stained with HLA-2.1/ CMV pp65-495 PE-tetramer. Then CMV pp65-495 is immunodominant peptide. (2) The use of HLA-peptide tetramer to quantify CMV specific T cells is valuable for monitoring and studying T-cell responses after allogeneic stem cell transplantation.
本课题的目的是利用制备的可溶性肽-HLA四聚体进行黑色素瘤抗原特异性细胞毒性T淋巴细胞的克隆和免疫反应监测的研究.肿瘤浸润性T淋巴细胞(TIL)620系(识别gp-100-209)用抗CD 8 mAb-PC 5和HLA-A2.1/gp 100 -209 PE-四聚体双重染色。通过荧光激活细胞分选(FACS)从TIL 620细胞系中分选出四聚体结合的CD 8 + T细胞(14%),并在体外扩增。培养的四聚体结合的CD 8 + T细胞对HLA-A2.1/gp 100-209 PE-四聚体的染色率为98.4%,并且释放的IFN-γ是原始TIL 620细胞系的7倍以上。利用HLA四聚体克隆抗原特异性活性T淋巴细胞成为可能.用突变试剂盒从HLA-A2.1中构建HLA-A2.6和A2. 7重链,用原核表达系统合成和纯化。产生HLA-A2.6/gp 100 -280四聚体,并通过gp 100 -280肽染色4- 5%的TIL 660(识别A2.1/gp-100-280表位)和7.97%的来自HLA-A2.6+健康供体的PBMC的诱导T细胞系。提示HLA-A2.1限制性黑色素瘤抗原肽可能与其他A2超型分子交叉识别.作为用HLA四聚体监测肿瘤特异性T淋巴细胞的模型,我们用荧光HLA-CMV肽四聚体监测异基因造血干细胞移植受体中CMV特异性T淋巴细胞的恢复,(1)用CMV pp 65 - 495肽在8例A2.1+健康供体的PMBC中诱导了7例CMV pp 65 - 495特异性T细胞,并用HLA-2.1/ CMV pp 65 -495 PE-四聚体染色。CMV pp 65 -495为免疫显性肽。(2)使用HLA-肽四聚体定量CMV特异性T细胞对于监测和研究异基因干细胞移植后的T细胞应答是有价值的。
项目成果
期刊论文数量(6)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Kawakami Yutaka: "Tumor Antigens Recognized by T cells and Antibodies. In "Human melanoma antigens recognized by CD8+ T cells""Taylor & Francis, New York. 47-74 (2003)
川上丰:“T 细胞和抗体识别的肿瘤抗原。在“CD8 T 细胞识别的人类黑色素瘤抗原”中”Taylor
- DOI:
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- 影响因子:0
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YUTAKA KAWAKAMI: "Isolation of a new melanoma antigen, MART-2, containing a mutated epitope recognized by autologous tumor infiltrating T lymphocytes"The Journal of Immunology. 116. 2871-2877 (2001)
YUTAKA KAWAKAMI:“分离出一种新的黑色素瘤抗原 MART-2,其中含有可被自体肿瘤浸润 T 淋巴细胞识别的突变表位”《免疫学杂志》。
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- 影响因子:0
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桜井 敏晴, 河上 裕: "抗原特異的T細胞を用いた癌、感染症の免疫療法"Molecular Medicine. 40(5). 582-589 (2003)
Toshiharu Sakurai、Yutaka Kawakami:“使用抗原特异性 T 细胞进行癌症和传染病的免疫治疗”,《分子医学》40(5)。
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- 影响因子:0
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Toshiharu Sakurai and Yutaka Kawakami: "Adoptive immunotherapy using CTL"Medical Science Digest. 28(7). 8-11 (2002)
Toshiharu Sakurai 和 Yutaka Kawakami:“使用 CTL 的过继免疫疗法”医学科学文摘。
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- 影响因子:0
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桜井 敏晴, 河上 裕: "細胞傷害性T細胞による養子免疫療法"Medical Science Digest. 28(7). 8-11 (2002)
Toshiharu Sakurai、Yutaka Kawakami:“使用细胞毒性 T 细胞的过继免疫疗法”《医学科学文摘》28(7)。
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SAKURAI Toshiharu其他文献
SAKURAI Toshiharu的其他文献
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{{ truncateString('SAKURAI Toshiharu', 18)}}的其他基金
Molecular link between stress response and cancer and its clinical implication
应激反应与癌症之间的分子联系及其临床意义
- 批准号:
17K09396 - 财政年份:2017
- 资助金额:
$ 2.3万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Novel molecular target in colorectal cancer and inflammatory bowel disease
结直肠癌和炎症性肠病的新分子靶点
- 批准号:
26460979 - 财政年份:2014
- 资助金额:
$ 2.3万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Translational research focusing on the regulation of proteasome by Gankyrin
Gankyrin对蛋白酶体调控的转化研究
- 批准号:
23590998 - 财政年份:2011
- 资助金额:
$ 2.3万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Elucidation of the resistance mechanisms of immunotherapy by the abnormal signal transduction in melanoma and development of methods to overcome it
阐明黑色素瘤中异常信号转导引起的免疫治疗耐药机制并开发克服该机制的方法
- 批准号:
21591445 - 财政年份:2009
- 资助金额:
$ 2.3万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
The study of immunosuppressive mechanisms induced by gastrointestinal cancer during epithelial-mesnchymal transition and clinical application of this target therapy
胃肠癌上皮间质转化过程中免疫抑制机制的研究及靶向治疗的临床应用
- 批准号:
18591484 - 财政年份:2006
- 资助金额:
$ 2.3万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Identification of novel autoantigens using human IgG in SCID mice implanted with lacrimal and salivary grands from patients with Sjogren's Syndrome
在植入干燥综合征患者泪腺和唾液腺的 SCID 小鼠中使用人 IgG 鉴定新型自身抗原
- 批准号:
16590324 - 财政年份:2004
- 资助金额:
$ 2.3万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Using solubel peptide-MHC tetramer, the trial of staining and cloning the melanoma antigen-specific cytotoxic lymphocytes
利用可溶性肽-MHC四聚体染色和克隆黑色素瘤抗原特异性细胞毒性淋巴细胞的试验
- 批准号:
11670849 - 财政年份:1999
- 资助金额:
$ 2.3万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Development and Characterization of Desmoglein-3 Specific T cells from Japanese Patients with Pemphigus Vulgaris
日本寻常型天疱疮患者 Desmoglein-3 特异性 T 细胞的发育和表征
- 批准号:
10670804 - 财政年份:1998
- 资助金额:
$ 2.3万 - 项目类别:
Grant-in-Aid for Scientific Research (C)