VIRAL MALIGNANT LYMPHOMAGENESIS IN X-IRRADIATED MICE

肿瘤进展是淋巴瘤发生的关键步骤。 这是一个 鲜为人知的过程 在小鼠淋巴瘤发生中，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 我们称之为T细胞淋巴母细胞瘤（TCLB），由正常的细胞组成， 二倍体，生长因子依赖性，在软琼脂中不克隆， 在体内皮下生长。 TCLB细胞是自刺激的， 稳定的肿瘤细胞 与此相反，我们发现的III期T细胞肿瘤 应称为T细胞淋巴瘤（TCL），含有能够自主分化的细胞。 可在软琼脂中定量克隆的生长物， 当接种在小鼠皮下时是致癌的。 III期TCL细胞 通常是15号染色体三体，遗传上高度稳定， 起源于胸腺 II期T细胞肿瘤具有 介于I期和III期肿瘤之间。 的本质 I期细胞到III期的进展包括逐渐的 生长因子独立性的发展。 I期细胞进展到 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。 我们将研究这种进展的机制。 将测试TCLB-和TCL细胞的细胞免疫球蛋白的表达。 致癌基因 从培养的肿瘤性T细胞中提取的Poly（A+）RNA 相I、II和III将在琼脂糖上分离后进行杂交 凝胶和“北方印迹”。 对14种不同的探针进行切口平移 克隆的癌基因、C型病毒长末端重复序列（LTR）和病毒 将使用包络序列。 表达水平以及基因 将寻找重排并与染色体异常相关 存在于II期肿瘤细胞的核型中， 三. 与T细胞相关的基因的表达和/或重排 增殖（如白细胞介素-2）也将进行研究， TCLB和TCL细胞在三个时期的表型差异。 在核型不同的TCL克隆中激活转化基因 将通过DNA介导的基因转移进行研究。 NIH 3 T3-和 将用从TCL分离的DNA转染淋巴样（TCLB）细胞 克隆 转化酶对限制性内切酶消化的敏感性 从不同的TCL克隆分离的DNA将用于描绘 存在于不同TCL克隆中的转化基因。 多克隆B细胞淋巴母细胞瘤（BCL B）已在B6小鼠中诱导， C型病毒复合体 我们会找到致病病毒 进行分子克隆。 在BCLB诱导中有活性的病毒基因将被 通过构建位点特异性病毒重组体进行研究。

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.