Kit ligand in synergy with interleukin-3 amplifies the erythropoietin- independent, globin-synthesizing progeny of normal human burst-forming units-erythroid in suspension cultures: physiologic implications

Although the proliferative effects of hematopoietic cytokines on erythroid progenitors are well known, parameters that influence the initiation of expression of specialized or lineage-restricted genes are not clear. We have studied the acquisition of erythroid-differentiative features from enriched populations of human early erythroid progenitors (burst-forming unit-erythroid BFUe) in suspension culture and the influence of several cytokines on this process. In suspension cultures containing no erythropoietin (Epo), we have found that kit ligand (KL) in synergy with interleukin-3 not only increases the proliferation of cells and of progenitors but also consistently amplifies a population of cells that contain globin within 1 week. Our experiments suggest that neither extraneously provided nor endogenously produced Epo is critical for the generation of globin-synthesizing cells. Globin-producing cells generated mostly from late BFUe or pre-CFUe with a CD34-/EP-1+ phenotype in this system do not all express a well-coordinated erythroid program accompanied by heme or glycophorin A expression and most die maintaining an immature state. Therefore, conditions that are responsible for initiation of globin expression in these cells are not sufficient to carry them to terminal maturation. The data point to an expanded target cell population for KL, as they suggest an influence of KL on survival and/or amplification of late erythroid cells previously thought to be influenced only by Epo. Our results in aggregate are of relevance to the physiology of normal erythropoiesis and the role of Epo and KL in the initial stages of lineage-restricted gene expression. In addition, they provide insight into the understanding of anemia in W and Steel mutants in which expansion of the late erythroid progenitor pool, normally dependent on the synergistic action of KL and Epo, is curtailed.