Bicknell D C, Williams D E, Broxmeyer H E
Department of Medicine, Indiana University School of Medicine, Indianapolis 46223.
Exp Hematol. 1988 Jan;16(1):88-91.
The gene product of the viral proto-oncogene v-fms, associated with the feline sarcoma virus (SM-FeSV), is a glycoprotein of 170 kd with associated tyrosine kinase activity. The murine c-fms proto-oncogene produces a similar glycoprotein of 165 kd and has been implicated as a similar, if not identical, molecule to the cell surface receptor of the hematopoietic growth factor CSF-1. Employing a v-fms probe in an in situ hybridization assay we examined the expression of c-fms transcripts in a population of purified granulocyte-macrophage progenitor cells (CFU-GM). Using this approach, which requires only 10-20 thousand cells, we demonstrate that 53% of the cells in the purified CFU-GM containing fraction (FR-28) express c-fms transcripts. In the presence of natural purified CSF-1, 51% +/- 3% of FR-28 cells form colonies or clusters in a semi-solid culture assay system. 94% of the colonies and clusters stimulated by CSF-1 were morphologically macrophage in nature. In addition, 84% +/- 4% of FR-28 cells formed colonies and clusters when stimulated with pokeweed mitogen spleen cell conditioned medium (PWMSCM) in a similar in vitro assay (35% granulocyte, 29% macrophage and 39% mixed granulocyte/macrophage colonies). These results indicate there is a correlation between the responsiveness of CFU-GM progenitor cells to CSF-1 and the expression of c-fms RNA.
与猫肉瘤病毒(SM-FeSV)相关的病毒原癌基因v-fms的基因产物是一种具有相关酪氨酸激酶活性的170kd糖蛋白。鼠类c-fms原癌基因产生一种类似的165kd糖蛋白,并且已被认为是造血生长因子CSF-1的细胞表面受体的相似分子(即便不是完全相同的分子)。我们运用v-fms探针进行原位杂交分析,检测了纯化的粒细胞-巨噬细胞祖细胞(CFU-GM)群体中c-fms转录本的表达。使用这种仅需1万至2万个细胞的方法,我们证明在纯化的含有CFU-GM的组分(FR-28)中,53%的细胞表达c-fms转录本。在天然纯化的CSF-1存在的情况下,51%±3%的FR-28细胞在半固体培养分析系统中形成集落或细胞簇。由CSF-1刺激形成的集落和细胞簇中,94%在形态上属于巨噬细胞。此外,在类似的体外分析中,当用商陆有丝分裂原脾细胞条件培养基(PWMSCM)刺激时,84%±4%的FR-28细胞形成集落和细胞簇(35%为粒细胞集落,29%为巨噬细胞集落,39%为粒细胞/巨噬细胞混合集落)。这些结果表明CFU-GM祖细胞对CSF-1的反应性与c-fms RNA的表达之间存在相关性。