IN VITRO CULTURE OF RADIATION INDUCED PRELEUKEMIC CELL

致白血病剂量的环磷酰胺在骨髓中诱导的初始改变 将进行辐射检查。 各种组织培养方法将 用于增殖和富集那些可以诱导 白血病过继转移实验。 这些改变的靶细胞将 检测其嗜亲性和重组病毒在 指示剂共培养测定。 细胞也将被检查， 内源性前病毒遗传元件的改变，通过Southern分析， DNA的限制酶。 同样，已知的一般位置 将在限制性内切酶中分析癌基因， 可以通过定量斑点杂交测定来测量。 其他转化相关蛋白和生长因子的表达 将通过代谢合成试验测定， 免疫沉淀/聚丙烯酰胺凝胶分析。 生长生物测定 将利用诸如白细胞介素2和3的因素来确定可能的 这些和其他生长因子的靶细胞表达的改变。 骨髓基质支持正常生长的能力，以及 还将检查辐射骨髓基质的生长因子， 部分和完全白血病诱导方案。 这些研究将导致确定专性与巧合 骨髓的改变，导致小鼠白血病的产生， 电离辐射

The initial alterations induced in bone marrow by leukemogenic doses of radiation will be examined. A variety of tissue culture methods will be utilized to propagate and enrich those target cells that can induce leukemia in adoptive transfer experiments. These altered target cells will be examined for their expression of ecotropic and recombinant virus in indicator co-cultivation assays. The cells will also be examined for alterations in endogenous proviral genetic elements by Southern analysis of restriction digests of their DNA. Similarly, the general location of known oncogenes will be analyzed in restriction digests and their expression will be measured by a quantitative dot blot hybridization assay. Expression of other transformation associated proteins and growth factors will be determined by metabolic synthesis assays using immunoprecipitation/-polyacrylamide gel analysis. Bioassays for growth factors such as interleukin 2 and 3 will be utilized to determine possible alterations in target cell expression of these and other growth factors. Marrow stromal capacity to support normal growth, and altered production of growth factors by radiated marrow stroma will also be examined during partial and complete leukemia induction protocols. Such studies will lead to a determination of obligate vs. coincidental alterations in bone marrow, resulting in generation of murine leukemia by ionizing radiation.