Induction of specific cellular immunity to EBV-positive T- and NK-lymphomas

诱导针对 EBV 阳性 T 和 NK 淋巴瘤的特异性细胞免疫

• 批准号: 12670802
• 负责人: KUZUSHIMA Kiyotaka
• 金额: $ 1.73万
• 依托单位: Aichi Cancer Center
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670802/
• 关键词: Epstein-Barr virus; CTL; HLA-A24; MHC-peptide tetramer; lymphoma; cytotoxic T lymphocyte; tetrameric MHN-peptide complex; HLA-A^*2402; LMP2; トランスレーショナルリサーチ; LMP1; TAT

EBV is associated with several malignant diseases, including a subset of lymphomas, nasopharyngeal carcinomas, Burkitt's lymphomas and immunoblastic lymphomas seen in immunocompromised hosts. We established an efficient approach for determination of CTL epitopes in the context of HLA A^*2402 molecules through multiple screenings, consisting of a computer-assisted algorithm, an in vitro MHC stabilization assay and an enzyme-linked immuno-spot (ELISPOT) assay. HLA A24 is the most frequently encountered HLA class I allele and the genotype is almost exclusively A^*2402. We first searched for candidate,peptides with the HLA A24-binding motif by a computer-assisted algorithm among EBV proteins reported to be targeted by specific CD8^+ T cells. We selected and synthesized 42 peptides after analyzing amino acid sequences of the 5 lytic and 6 latent cycle proteins. All of them functionally stabilized HLA A^*2402 molecules which had been expressed on the peptide transporter-deficient cell line T2. Next is screening of the peptides by ELISPOT assay for their recognition by bulk EBV-specific CD8^+ T cells, established from PBMCs of EBV-immune donors positive for HLA A24.-typing. We have identified 2 possible major lytic cycle EBV-specific CTL epitopes from the amino acid sequences of BRLF1 and BMLF1 and 1 from LMP2 presented by HLA A^*2402 molecules. The newly identified peptides should prove useful for detection and/or study of EBV-specific CD8^+ T cell responses among populations positive for HLA A^*2402. We produced HLA A^*2402tetramers-incorporating the 3 peptides and 2 known epitope peptide, one derived from EBNA3A (RYSIFFDYM), and the other derived from EBNA3B (TYSAGIVQI). All the 5 tetramers successfully stained various proportions of EBV-specific CD8+ T cells. The newly identified peptides should prove useful for detection and/or study of EBV-specific CD8^+ T cell responses among populations positive for HLAA^*2402.

EBV与几种恶性疾病有关，包括在免疫功能低下的宿主中看到的淋巴瘤亚群、鼻咽癌、伯基特淋巴瘤和免疫母细胞淋巴瘤。我们通过计算机辅助算法、体外MHC稳定试验和酶联免疫斑点（ELISPOT）试验等多种筛选方法，建立了在HLA A^*2402分子背景下测定CTL表位的有效方法。HLA A24是最常见的HLA I类等位基因，其基因型几乎完全为A^*2402。我们首先通过计算机辅助算法在被报道为特异性CD8^+ T细胞靶向的EBV蛋白中搜索具有HLA a24结合基序的候选肽。通过分析5个裂解蛋白和6个潜伏循环蛋白的氨基酸序列，我们选择并合成了42个多肽。它们都能稳定在肽转运蛋白缺陷细胞系T2上表达的HLA A^*2402分子。接下来，通过ELISPOT检测筛选这些肽，以供大量ebv特异性CD8^+ T细胞识别，这些细胞来自HLA a24分型阳性的ebv免疫供者的PBMCs。我们从BRLF1和BMLF1的氨基酸序列和HLA A^*2402分子呈现的LMP2的氨基酸序列中鉴定出2个可能的主要裂解周期ebv特异性CTL表位。新鉴定的多肽将被证明可用于检测和/或研究HLA A^*2402阳性人群中ebv特异性CD8^+ T细胞反应。我们制作了HLA A^*2402四聚体，包含3个肽和2个已知的表位肽，一个来自EBNA3A (RYSIFFDYM)，另一个来自EBNA3B （TYSAGIVQI）。5种四聚体均成功染色不同比例的ebv特异性CD8+ T细胞。新鉴定的肽将被证明可用于检测和/或研究HLAA^*2402阳性人群中ebv特异性CD8^+ T细胞反应。

项目成果

葛島清隆: "EBV関連lymphoproliferative disease:診断と治療の最近の動向."化学療法の領域. 15・9. 49-53 (1999)

Kiyotaka Katsurashima：“EBV 相关淋巴增殖性疾病：化疗的最新趋势”15・9（1999）。

Tsuge I, Morishima T, Kimura H, Kuzushima K, Matsuoka H: "Impaired cytotoxic T lymphocyte response to Epstein-Barr virus-infected NK cells in patients with severe chronic active EBV infection"J Med Virol. 64(2). 141-148 (2001)

Tsuge I、Morishima T、Kimura H、Kuzushima K、Matsuoka H：“严重慢性活动性 EBV 感染患者对 Epstein-Barr 病毒感染的 NK 细胞的细胞毒性 T 淋巴细胞反应受损”J Med Virol。

葛島清隆: "細胞性免疫からみたEpstein-Barr virus感染症"ウイルス. 51(1). 43-49 (2001)

Kiyotaka Katsurashima：“从细胞介导的免疫角度观察 Epstein-Barr 病毒感染”51(1) (2001)。

Fujii K: "The Epstein-Barr virus pol catalytic subunit physically interacts with the BBLF4-BSLF1-BBLF2/3 complex"J.Virol. 74・6. 199-201 (2000)

Fujii K：“Epstein-Barr 病毒 pol 催化亚基与 BBLF4-BSLF1-BBLF2/3 复合物发生物理相互作用”J. Virol 74・6 (2000)。

Kuzushima K: "Longitudinal dynamics of EBV-specific CTL during the posttransplant lymphoproliferative disorder"Journal of Infections Disease. 182. 937-940 (2000)

Kuzushima K：“移植后淋巴增殖性疾病期间 EBV 特异性 CTL 的纵向动态”感染疾病杂志。