REGULATION OF INTERLEUKIN 2 GENE EXPRESSION IN LYMPHOID AND NONLYMPHOID CELLS

这个项目的重点是了解基本的控制 人白细胞介素2的表达机制 （IL-2）基因。 IL-2基因表达受到严格控制， 仅限于T细胞的抗原刺激。 为了研究 细胞特异性调节IL-2基因，我们已经转染， 人IL-2基因导入小鼠NIH-3 T3成纤维细胞、人HeLa细胞 和小鼠BFS淋巴瘤T细胞，并随后分析稳定 含有导入的基因的细胞， 表情 所有三种细胞系都被发现表达外源性 基因组成，虽然在低水平。 这一点得到了证明， 免疫可沉淀蛋白和生物活性两者存在 细胞上清液中IL-2的特征，以及mRNA 在与人IL-2 cDNA杂交的细胞提取物中。 令人好奇的是，所观察到的mRNA种类比人类预期的要大。 IL-2，似乎含有内含子序列。 治疗 佛波醇肉豆蔻酸酯乙酸酯和钙离子载体A23187产生 BFS中转染的人IL-2基因表达增加 细胞，而不是3 T3或HeLa细胞。 我们得出结论，人IL- 2基因转染淋巴和非淋巴细胞， 组成型表达，而其蛋白激酶C/钙- 介导的诱导型表达仍然局限于淋巴样细胞。 我们的数据是一致的发生的不平衡， 正负调控DNA序列相互作用 调节IL-2基因表达，并提供一个有用的实验 为他们的调查做榜样。

This project is focused on understanding the basic control mechanisms responsible for expression of the human interleukin 2 (IL-2) gene. IL-2 gene expression is tightly controlled and limited to antigenic stimulation of T cells. In order to study the cell specific regulation of the IL-2 gene we have transfected the human IL-2 gene into mouse NIH-3T3 fibroblasts, human HeLa cells and mouse BFS lymphoma T cells and subsequently analyzed stable transformant cells containing the introduced gene for its expression. All three cell lines were found to express the foreign gene constitutively albeit at low levels. This was evidenced by the presence of both immunoprecipitable protein and bioactivity characteristic of IL-2 in cellular supernatants, as well as mRNA in cellular extracts which hybridized to a human IL-2 cDNA. Curiously, the mRNA species seen was larger than expected for human IL-2 and appears to contain intronic sequences. Treatment with phorbol myristate acetate and the calcium ionophore A23187 resulted in increased expression of the transfected human IL-2 gene in BFS cells but not in 3T3 or HeLa cells. We conclude that the human IL- 2 gene transfected into lymphoid and nonlymphoid cells is constitutively expressed while its protein kinase C/calcium- mediated inducible expression remains restricted to lymphoid cells. Our data are consistent with the occurrence of an imbalance of the interplay of positive and negative regulatory DNA sequences modulating IL-2 gene expression and provide a useful experimental model for their investigation.