Development of novel immunogene therapy for hematological malignancies

血液系统恶性肿瘤新型免疫基因疗法的开发

• 批准号: 12557081
• 负责人: YASUKAWA Masaki
• 金额: $ 7.68万
• 依托单位: Ehime University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557081/
• 关键词: Leukemia; Malignant lymphoma; Immunotherapy; Gene therapy; Cytotoxic T lymphocytes; 造血器腫瘍; 細胞免疫療法; WT1; テロメラーゼ; 細胞障害性T細胞; 自殺遺伝子; 不死化細胞; HLA; hTERT

This study was performed to develop the novel immunotherapy for hematological malignancies. The data obtained from the series of experiments are as follows.1) A novel WT1-derived peptide-specific CD8^+ CTL line, designated NIM-1 was established. NIM-1 lysed HLA-A24-positive leukaemia cells, but not HLA-A24-negative leukaemia cells or normal cells.2) A WT1-specific, HLA-A24-restricted CTL clone (designated TAK-1) exhibited cytotoxicity against lung cancer cell lines bearing HLA-A24 but did not lyse cells lacking this HLA. Adoptive transfer of TAK-1 into nude mice that had been engrafted with an HLA-A24-positive lung cancer cell line resulted in inhibition of the cancer cell growth and prolonged survival. These findings strongly suggest that WT1 is a universal tumor-associated antigen and that WT1 -targeting immunotherapy offers a potentially effective treatment option for lung cancer as well as leukemia.3) Immature dendritic cells (DCs) were loaded with leukemia cells with t(6 ; 9) or t … More (9 ; 22) and then cocultured with the dek-can fusion peptide-specific or the bcr abl fusion peptide-specific CD4^+ T-lymphocyte clone. The dek-can peptide-specific and bcr-abl peptide-specific CD4^+ T-lymphocyte clones produced interferon-γ (IFN-γ) when they were cocultured with HLA-DR-matched but not with mismatched DCs which had been loaded with apoptotic as well as necrotic leukemia cells with t(6 ; 9) and t(9 ; 22), respectively. These data indicate that the acute myelogenous leukemia-associated fusion protein, dek-can and chronic myelogenous leukemia-associated fusion protein, bcr-abl, are both processed and presented by DCs to the fusion peptide-specific CD4^+ T lymphocytes.4) In order to clarify the roles of perform in antigen-specific cytotoxicity mediated by human CD4^+ CTLs, antigen-specific human CD4^+ T-lymphocyte clones were established from a patient with hereditary perforin deficiency and their cytotoxic activities were investigated. The data demonstrated that perforin-negative CD4^+ CTLs can exert cytotoxicity against Fas-sensitive target cells ; however, perforin plays essential roles in antigen-specific cytotoxicity mediated by human CD4^+ as well as CD8^+ CTLs. Less

本研究旨在开发一种新的恶性血液病免疫治疗方法。1）建立了一种新的WT 1多肽特异性CD 8 ^+ CTL细胞系NIM-1。NIM-1裂解HLA-A24阳性白血病细胞，但不裂解HLA-A24阴性白血病细胞或正常细胞。2）WT 1特异性、HLA-A24限制性CTL克隆（命名为TAK-1）对携带HLA-A24的肺癌细胞系表现出细胞毒性，但不裂解缺乏该HLA的细胞。将TAK-1连续转移至已植入HLA-A24阳性肺癌细胞系的裸鼠体内，可抑制癌细胞生长并延长存活期。这些发现有力地表明WT 1是一种通用的肿瘤相关抗原，WT 1靶向免疫治疗为肺癌和白血病提供了一种潜在有效的治疗选择。 ...更多信息 （9 ; 22），然后与dek-can融合肽特异性或bcr abl融合肽特异性CD 4 ^+ T淋巴细胞克隆共培养。当dek-can肽特异性和bcr-abl肽特异性CD 4 ^+ T淋巴细胞克隆与HLA-DR匹配的DC共培养时，它们产生干扰素-γ（IFN-γ），而与不匹配的DC共培养时，则不产生干扰素-γ（IFN-γ），而不匹配的DC分别负载了t（6 ; 9）和t（9 ; 22）的凋亡和坏死白血病细胞。这些数据表明，急性髓细胞白血病相关融合蛋白dek-can和慢性髓细胞白血病相关融合蛋白bcr-abl均被DCs加工并提呈给融合肽特异性CD 4 ^+ T淋巴细胞。4）为了阐明perform在人CD 4 ^+ CTL介导的抗原特异性细胞毒作用中的作用，从遗传性穿孔素缺乏症患者中建立了抗原特异性人CD 4 ^+ T淋巴细胞克隆，并研究了它们的细胞毒活性。这些数据表明，穿孔素阴性的CD 4 ^+ CTL可以对Fas敏感的靶细胞产生细胞毒性;然而，穿孔素在由人CD 4 ^+和CD 8 ^+ CTL介导的抗原特异性细胞毒性中起着重要作用。少

项目成果

Azuma, T., et al.: "Identification of a novel WT1-derived peptide which induces HLA-A24-restricted anti-leukaemia cytotoxic T lymphocytes"British Journal of Haematology. 116. 601-603 (2002)

Azuma，T.，等人：“诱导 HLA-A24 限制性抗白血病细胞毒性 T 淋巴细胞的新型 WT1 衍生肽的鉴定”英国血液学杂志。

Yasukawa M., et al: "Analysis of HLA-DRB1 alleles in Japanese patients with chronic myelogenous leukemia"American Journal of Hematology. 63. 99-101 (2000)

Yasukawa M.等人：“日本慢性粒细胞白血病患者的HLA-DRB1等位基因分析”美国血液学杂志。

Hamada, M., Yakushijin, Y., Watanabe, L., Kakimoto, M., Yasukawa, M. and Fujita, S.: "Aurora2/BTAK/STKI5 is involved in cell-cycle checkpoint and cell survival of aggressive non-Hodgkin's lymphoma"Br.J.Haematol.. (in press).

Hamada, M.、Yakushijin, Y.、Watanabe, L.、Kakimoto, M.、Yasukawa, M. 和 Fujita, S.：“Aurora2/BTAK/STKI5 参与侵袭性非-细胞周期检查点和细胞存活。

Kakimoto, M., Hasegawa, A., Fujita, S. and Yasukawa, M.: "Phenotypic and functional alterations of dendritic cells induced by human herpesvirus 6 infection"J.Virol.. 76. 10338-10345 (2002)

Kakimoto, M.、Hasekawa, A.、Fujita, S. 和 Yasukawa, M.：“人疱疹病毒 6 感染诱导的树突状细胞的表型和功能改变”J.Virol.. 76. 10338-10345 (2002)

Hasegawa, A., et al.: "Transcriptional down-regulation of CXCR4 induced by impaired association of YY1 with c-Myc in HHV-6-infected cells"Journal of Immunology. 166. 1125-1131 (2001)

Hasekawa, A. 等人：“HHV-6 感染细胞中 YY1 与 c-Myc 的关联受损导致 CXCR4 转录下调”《免疫学杂志》。