VIRAL MALIGNANT LYMPHOMAGENESIS IN X-IRRADIATED MICE

X 射线照射小鼠中的病毒恶性淋巴瘤发生

• 批准号: 3171916
• 负责人: Martin nmi Haas
• 金额: $ 17.87万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-05-01 至 1988-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3171916
• 关键词: T lymphocyte; X ray; gene expression; genetic manipulation; leukocyte activation /transformation; lymphoma; natural gene amplification; neoplasm /cancer genetics; oncogenes; radiation related neoplasm /cancer; tissue /cell culture; transforming growth factors; viral carcinogenesis; virus DNA; virus genetics

Tumor progression is a critical step in lymphomagenesis. It is a little-understood process. In murine lymphomagenesis we have found three distinct phases of T cell neoplasms: T cell neoplasms of phase I, which we shall call T cell lymphoblastoma (TCLB), consist of cells that are normal diploid, growth factor dependent, do not clone in soft agar, and do not grow under the skin in vivo. TCLB cells are autostimulating, genetically stable tumor cells. In contrast T cell neoplasms of phase III, which we shall call T cell lymphomas (TCL), contain cells capable of autonomous growth which can be cloned quantitatively in soft agar and which are tumorigenic when inoculated under the skin of mice. Phase III TCL cells are often trisomic for chromosome 15 and highly stable genetically; they originate in the thymus. T cell neoplasms of phase II have properties intermediate between neoplasms of phase I and phase III. The essence of progression of phase I cells to phase III consists of the gradual development of growth factor independence. Phase I cells progress to phase II and possibly to phase III cells, a process accompanied by increasing autonomy of the cells. We will study the mechanism of this progression. TCLB- and TCL cells will be tested for the expression of cellular oncogenes. Poly (A+) RNA extracted from cultured neoplastic T cells of phases I, II, and III, will be hybridized following separation on agorose gels and "Northern blotting". Nick-translated probes made to 14 different cloned oncogenes, C-type viral long terminal repeats (LTR) and viral envelope sequences will be used. The level of expression as well as gene rearrangements will be sought and correlated with chromosome abnormalities that are present in the karyotypes of the neoplastic cells of phases II and III. Expression and/or rearrangement of genes associated with T cell proliferation (e.g. interleukin-2) will also be studied, as will the phenotypic differences among TCLB and TCL cells of the three phases. Activation of transforming genes in karyotypically different TCL clones will be studied by means of DNA-mediated gene transfer. NIH 3T3- and lymphoid (TCLB) cells will be transfected with DNA isolated from TCL clones. Sensitivity to digestion with restriction enzymes of transforming DNAs isolated from different TCL clones will be used to delineate the transforming genes present in different TCL clones. Polyclonal B cell lymphoblastomas (BCLB) have been induced in B6 mice by a potent C-type virus complex. The pathogenic virus will be sought and will be molecularly cloned. Viral genes active in the induction of BCLB will be studied by constructing site-specific virus recombinants.

肿瘤进展是淋巴肿大发生的关键步骤。这是一个 鲜为人知的过程。在小鼠淋巴增生症中，我们发现了三种 T细胞肿瘤的不同阶段：I期T细胞肿瘤，我们 称为T细胞淋巴母细胞瘤(TCLB)，由正常细胞组成 二倍体，依赖于生长因子，不在软琼脂中克隆，也不 在活体的皮肤下生长。TCLB细胞在遗传上具有自我刺激功能 稳定的肿瘤细胞。相比之下，T细胞肿瘤是III期的，我们 称为T细胞淋巴瘤(TCL)，含有自主性细胞 可以在软琼脂中定量克隆的生长， 当接种在小鼠的皮肤下时会产生肿瘤。III期TCL细胞 通常是15号染色体的三体，在遗传上高度稳定；他们 起源于胸腺。II期T细胞肿瘤具有某些特性 介于I期和III期肿瘤之间的中间体。 I期细胞向III期细胞的进展包括渐进的 发展对生长因子的独立性。第一阶段细胞进展到阶段 II期和可能的III期细胞，这一过程伴随着增加 细胞的自主性。我们将研究这一进展的机制。 将对TCLB-和TCL细胞进行细胞表达检测 致癌基因。从培养的人肿瘤T细胞中提取Poly(A+)RNA 第一阶段、第二阶段和第三阶段将在琼脂糖上分离后进行杂交 凝胶和“Northern印迹”。尼克翻译的探针制成了14个不同的 癌基因克隆、C型病毒长末端重复序列与病毒 将使用包络序列。表达水平和基因 将寻求与染色体异常相关的重排。 存在于II期和第II期肿瘤细胞的核型中 三、T细胞相关基因的表达和/或重排 还将研究增殖(如白细胞介素2)，以及 三个时相TCLB和TCL细胞的表型差异。 核型不同TCL克隆中转化基因的激活 将通过DNA介导的基因转移进行研究。NIH 3T3-和 淋巴样细胞(TCLB)将用从TCL中提取的DNA进行转基因 克隆人。转化限制性内切酶对消化的敏感性 从不同TCL克隆中分离出来的DNA将被用来描述 转化基因存在于不同的TCL克隆中。 在B6小鼠体内诱导了多克隆性B细胞淋巴母细胞瘤。 强大的C型病毒复合体。致病病毒将被寻找并将 被分子克隆。在BCLB诱导中活跃的病毒基因将是 通过构建位点特异性病毒重组体进行研究。