Immunogene therapy of cancer and virus infections using immortalized T-cell clones

使用永生化 T 细胞克隆对癌症和病毒感染进行免疫基因治疗

• 批准号: 11670449
• 负责人: YASUKAWA Masaki
• 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: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670449/
• 关键词: Cancer; Leukemia; Immunotherapy; Gene therapy; WT1; Telomerase; HLA; Cytotoxic T cells; がん免疫; 免疫遺伝子治療; 白血病; ウイルス感染症

1. The WT1 gene encodes a zinc finger transcription factor, which is preferentially expressed in acute leukemia cells and chronic myelogenous leukemia cells in blast crisis, but not in most normal cells. These findings strongly suggest that WT1 is a potential target of immunotherapy for human leukemia. We have established a CD8+ cytotoxic Tlymphocyte (CTL) clone, designated TAK-1, which is specific for a WT1-derived 9-mer peptide consisting of HLA-A24-binding anchor motifs. TAK-1 lysed both HLA-A24-positive allogeneic cells and autologous cells that were loaded with a WT1-derived peptide. TAK-1 was cytotoxic to HLA-A24-positive leukemia cells, but not to HLA-A24-positive lymphoma cells that did not express WT1, to HLA-A24-negative leukemia cells, or to HLA-A24-positive normal cells. Treating leukemia cells with an antisense oligonucleotide complementary to WT1 reduced TAK-1-mediated cytotoxicity. TAK-1 did not inhibit colony formation of HLA-A24-positive normal bone marrow cells.2. Because hTERT is preferentially expressed in malignant cells but not in normal cells, it is considered to be a potential target for cancer immunotherapy. We examined hTERT-derived peptides carrying motifs for HLA-A24 (HLA-A^*2402) for their capacity to elicit anti-leukemia cytotoxic Tlymphocytes (CTLs). Two of the 5 peptides tested, VYAETKHFL and VYGFVRACL, appeared capable of generating hTERT peptide-specific and HLA-A24-restricted CTLs. The CD8+ CTL clones specific for these hTERT peptides exerted cytotoxicity against leukemia cells in an HLA-A24-restricted manner. The cytotoxicity was inhibited by adding hTERT peptide-loaded autologous-LCLs, suggesting that hTERT peptide is naturally processed and expressed on leukamia cells, and can be lysed by these CTLs. Taken together with the currently identified HLA-A2-restricted CTL epitopes derived from hTERT, identification of CTL epitopes presented by HLA-A24 extends the feasibility of immunotherapy for leukemia using hTERT-derived peptides.

1. WT 1基因编码锌指转录因子，其优先在急性白血病细胞和处于急变期的慢性髓性白血病细胞中表达，而在大多数正常细胞中不表达。这些发现有力地表明WT 1是人类白血病免疫治疗的潜在靶点。我们已经建立了一个CD 8+细胞毒性T淋巴细胞（CTL）克隆，命名为TAK-1，它是特异性的WT 1衍生的9-mer肽组成的HLA-A24结合锚基序。TAK-1裂解HLA-A24阳性同种异体细胞和负载WT 1衍生肽的自体细胞。TAK-1对HLA-A24阳性白血病细胞具有细胞毒性，但对不表达WT 1的HLA-A24阳性淋巴瘤细胞、HLA-A24阴性白血病细胞或HLA-A24阳性正常细胞无细胞毒性。用与WT 1互补的反义寡核苷酸处理白血病细胞可降低TAK-1介导的细胞毒性。TAK-1不抑制HLA-A24阳性正常骨髓细胞的集落形成。2.由于hTERT在恶性肿瘤细胞中优先表达，而在正常细胞中不表达，因此它被认为是癌症免疫治疗的潜在靶点。我们检测了携带HLA-A24（HLA-A^*2402）基序的hTERT衍生肽引发抗白血病细胞毒性T淋巴细胞（CTL）的能力。所测试的5种肽中的两种，VYAETKHFL和VYGFVRACL，似乎能够产生hTERT肽特异性和HLA-A24限制性CTL。这些hTERT肽特异性的CD 8 + CTL克隆以HLA-A24限制性方式对白血病细胞产生细胞毒性。加入负载hTERT肽的自体LCL可抑制细胞毒性，表明hTERT肽在白血病细胞上天然加工和表达，并可被这些CTL裂解。与目前鉴定的源自hTERT的HLA-A2限制性CTL表位一起，鉴定由HLA-A24呈递的CTL表位扩展了使用hTERT衍生肽免疫治疗白血病的可行性。

项目成果

Yasukawa, M., Hasegawa, A., Sakai, I., Ohminami, H., Arai, J., Kaneko, S., Yakushijin, Y., Maeyama, K., Nakashima, H., Arakaki, R.and Fujita, S.: "Down-regulation of CXCR4 by human herpesvirus 6 (HHV-6) and HHV-7."J.Immunol. 162. 5417-5422 (1999)

安川，M.，长谷川，A.，酒井，I.，大南，H.，荒井，J.，金子，S.，药师人，Y.，前山，K.，中岛，H.，荒崎，R.和

Yasukawa,M.,et al.: "Down-regulation of CXCR4 by human herpesvirus 6(HHV-6)and HHV-7"Journal of Immunology. 162. 5417-5422 (1999)

Yasukawa, M., et al.：“人疱疹病毒 6 (HHV-6) 和 HHV-7 下调 CXCR4”免疫学杂志。

安川正貴: "白血病・リンパ腫・骨髄腫"押味和夫 編集 中外医学社. 81-98 (2000)

安川正孝：“白血病、淋巴瘤、骨髓瘤”，大岛和夫编辑，中外医学社 81-98 (2000)。

Hasegawa, A., Yasukawa, M., Sakai, I.and Fujita, S.: "Transcriptional down-regulation of CXCR4 induced by impaired association of transcription regulator YY1 with c-Myc in human herpesvirus 6-infected cells."J.Immunol. 166. 1125-1131 (2001)

Hasekawa, A.、Yasukawa, M.、Sakai, I. 和 Fujita, S.：“在人疱疹病毒 6 感染的细胞中，转录调节因子 YY1 与 c-Myc 的关联受损，导致 CXCR4 的转录下调。”

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。