Miller M, Kennewell A, Takayama Y, Bruskin A, Bishop J M, Johnson G, Symonds G
Children's Medical Research Foundation, Camperdown, Sydney, Australia.
Oncogene. 1990 Aug;5(8):1125-31.
Avian erythroblastosis virus (AEV) is a replication-defective retrovirus that transforms erythroid and fibroblast cells in vitro and in vivo. The transforming ability of AEV is due primarily to the oncogene v-erb-B. A recombinant murine retrovirus has been constructed by inserting a chimeric gag-v-erb-B gene into a Moloney murine leukemia virus based vector. This retrovirus was used to examine v-erb-B-induced transformation of murine hematopoietic cells. Infection of murine primary fetal liver, adult bone marrow or adult spleen cells with the recombinant virus generated large hemoglobinized erythroid colonies in the absence of exogenous growth factors. Generation of such colonies usually requires the presence of erythropoietin (Epo) and interleukin-3 (IL-3). These growth-factor independent colonies were shown to be derived from early (BFU-E) and not late (CFU-E) erythroid progenitor cells, and the effect was not attributable to growth factors elicited by the virus-producing cell lines. In order to confirm that the recombinant virus was responsible for this transformation of BFU-E to growth factor independence, bone marrow cells from post 5-fluorouracil treated mice were infected and used to repopulate lethally-irradiated mice. Growth factor-independent BFU-E were obtained in up to 30% of day-13 spleen colonies and it was shown by DNA analysis that cells from these colonies contained integrated provirus. Our results indicate that v-erb-B transforms early erythroid progenitors to growth factor independent growth and subsequent differentiation to erythrocytes -a process that normally requires Epo plus either IL-3 or granulocyte-macrophage colony stimulating factor (GM-CSF).
禽成红细胞增多症病毒（AEV）是一种复制缺陷型逆转录病毒，可在体外和体内转化红细胞和成纤维细胞。AEV的转化能力主要归因于癌基因v-erb-B。通过将嵌合的gag-v-erb-B基因插入基于莫洛尼鼠白血病病毒的载体中，构建了一种重组鼠逆转录病毒。该逆转录病毒用于检测v-erb-B诱导的鼠造血细胞转化。用重组病毒感染鼠原代胎肝、成年骨髓或成年脾细胞，在无外源性生长因子的情况下产生了大量血红蛋白化的红细胞集落。通常需要存在促红细胞生成素（Epo）和白细胞介素-3（IL-3）才能产生这样的集落。这些不依赖生长因子的集落被证明源自早期（BFU-E）而非晚期（CFU-E）红细胞祖细胞，且该效应不归因于病毒产生细胞系引发的生长因子。为了证实重组病毒是BFU-E向不依赖生长因子转化的原因，对5-氟尿嘧啶处理后的小鼠的骨髓细胞进行感染，并用于重建经致死性照射的小鼠。在第13天的脾集落中，高达30%获得了不依赖生长因子的BFU-E，并且通过DNA分析表明，这些集落中的细胞含有整合的前病毒。我们的结果表明，v-erb-B将早期红细胞祖细胞转化为不依赖生长因子的生长，并随后分化为红细胞——这一过程通常需要Epo加上IL-3或粒细胞-巨噬细胞集落刺激因子（GM-CSF）。
