Nocka K, Buck J, Levi E, Besmer P
Molecular Biology, Sloan Kettering Institute, New York, NY 10021.
EMBO J. 1990 Oct;9(10):3287-94. doi: 10.1002/j.1460-2075.1990.tb07528.x.
The c-kit proto-oncogene encodes a transmembrane tyrosine kinase receptor for an unidentified ligand and is allelic with the murine white-spotting locus (W). W mutations affect melanogenesis, gametogenesis and hematopoiesis during development and in adult life. Cellular targets of W mutations in hematopoiesis include distinct cell populations in the erythroid and mast cell lineages as well as stem cells. In the absence of interleukin-3 (IL-3) mast cells derived from normal mice but not from W mutant mice can be maintained by co-culture with 3T3 fibroblasts. Based on the defective proliferative response of W mast cells in the 3T3 fibroblast co-culture system it had been proposed that fibroblasts produce the c-kit ligand. We have used a mast cell proliferation assay to purify a 30 kd protein, designated KL, from conditioned medium of Balb/3T3 fibroblasts to apparent homogeneity. KL stimulates the proliferation of normal bone marrow derived mast cells but not mast cells from W mice, although both normal and mutant mast cells respond similarly to IL-3. Connective tissue-type mast cells derived from the peritoneal cavity of normal mice were found to express a high level of c-kit protein on their surface and to proliferate in response to KL. The effect of KL on erythroid progenitor cells was investigated as well. In combination with erythropoietin, KL was found to stimulate early erythroid progenitors (BFU-E) from fetal liver and spleen cells but not from bone marrow cells of adult mice and from fetal liver cells of W/W mice.(ABSTRACT TRUNCATED AT 250 WORDS)
原癌基因c-kit编码一种针对未知配体的跨膜酪氨酸激酶受体,与小鼠的白斑位点(W)等位。W突变在发育过程和成年期影响黑色素生成、配子发生和造血作用。造血过程中W突变的细胞靶点包括红系和肥大细胞系中的不同细胞群体以及干细胞。在缺乏白细胞介素-3(IL-3)的情况下,源自正常小鼠而非W突变小鼠的肥大细胞可通过与3T3成纤维细胞共培养来维持。基于W肥大细胞在3T3成纤维细胞共培养系统中的增殖反应缺陷,有人提出成纤维细胞产生c-kit配体。我们利用肥大细胞增殖试验从Balb/3T3成纤维细胞的条件培养基中纯化出一种30kd的蛋白,命名为KL,达到明显的均一性。KL刺激正常骨髓来源的肥大细胞增殖,但不刺激W小鼠的肥大细胞,尽管正常和突变的肥大细胞对IL-3的反应相似。发现源自正常小鼠腹腔的结缔组织型肥大细胞在其表面表达高水平的c-kit蛋白,并对KL产生增殖反应。还研究了KL对红系祖细胞的作用。发现KL与促红细胞生成素联合使用时,可刺激来自胎肝和脾细胞的早期红系祖细胞(BFU-E),但不刺激成年小鼠骨髓细胞和W/W小鼠胎肝细胞的早期红系祖细胞。(摘要截短于250字)