Development of New diagnostic methods and treatments against EB virus as a target in nasal NK/T cell lymphoma

(1)病毒学分析采用cDNA芯片技术分析鼻腔NK/T细胞淋巴瘤细胞系，探讨细胞因子的作用。IL-9在EB病毒阳性鼻部NK/T细胞淋巴瘤细胞株中表达，在阴性鼻部NK/T细胞淋巴瘤细胞株中不表达。此外，IL-9似乎在细胞系中通过自分泌机制促进了细胞的增殖。我们分析了鼻部NK/T细胞淋巴瘤组织中LMP1和LMP2A基因的突变。在组织中发现LMP1缺失和LMP2A突变。我们通过抗体阵列分析鼻部NK/T细胞淋巴瘤细胞系的趋化因子。IP-10在鼻腔NK/T细胞淋巴瘤细胞系中表达。另外，IP-10似乎通过自分泌机制参与了细胞的迁移。(2)建立新的诊断方法我们建立了一种实时PCR检测鼻腔NK/T细胞淋巴瘤患者血清EBV DNA载量的方法。血清EBV DNA载量对本病的诊断和预测患者预后及复发有重要意义。(3)新治疗方法的发展我们开发的经动脉化疗和放疗治疗鼻NK/T细胞淋巴瘤有效且安全。利用LMP1肽培养辅助性T淋巴细胞。T细胞识别一种LMP1 159-175的多肽，它具有混杂的LMP1和限制性HLA-DR9、DR53和DR15的表位。此外，T细胞杀死淋巴瘤细胞。

(1) Virological analysisWe analyzed nasal NK/T cell lymphoma cell line to investigate the role of cytokines by cDNA microarray. IL-9 was expressed in EB virus positive nasal NK/T cell lymphoma cell line but not negative cell lines. In addition, it seems that IL-9 has contributed for the cell proliferation by autocrine mechanism in the cell line. s.We analyzed gene mutation of LMP1 and LMP2A in nasal NK/T cell lymphoma tissues. Deletion of LMP1 and mutations of LMP2A were discovered in the tissuesWe analyzed chemokines in nasal NK/T cell lymphoma cell line by the antibody array. IP-10 was expressed in nasal NK/T cell lymphoma cell line. In addition, it seems that IP-10 has contributed for the cell migration by autocrine mechanism.(2) Development of new diagnostic methodsWe developed a real time PCR method to measure EBV DNA load in sera of nasal NK/T cell lymphoma patients. Serum EBV DNA load was useful for diagnosis of this disease and to predict patient's prognosis and recurrence.(3) Development of new treatment methodsIntra-arterial chemotherapy and radiotherapy we developed was useful and safety to treat nasal NK/T cell lymphoma.Helper T lymphocytes were developed using LMP1 peptides. The T cells recognize a peptide (LMP1 159-175) with promiscuous epitopes of LMP1 and restricted HLA-DR9, DR53 and DR15. In addition, the T cells killed lymphoma cells.