Analysis of interleukin 6 receptor and gp130 expressions and proliferative capability of human CD34(+) cells

We recently demonstrated that stimulation of gp130 by a combination of soluble interleukin 6 receptor (sIL-6R) and IL-6 but not IL-6 alone significantly stimulates the ex vivo expansion of primitive hematopoietic progenitors and the generation of erythroid cells from human CD34(+) cells in the presence of stem cell factor (SCF). Here, we show that gp130 is found low positively on most CD34(+) cells, whereas IL-6R is expressed on only 30-50% of these cells. Although most of the colonies generated from FACS(R)-sorted CD34(+)IL-6R(+) cells were granulocyte/macrophage (GM) colonies, CD34(+)IL-6R(-) cells gave rise to various types of colonies, including erythroid bursts, GM, megakaryocytes, and mixed colonies in methylcellulose culture with a combination of IL-6, sIL-6R, and SCF. Similar results were obtained in culture supplemented with a combination of IL-3, IL-6, SCF, granulocyte colony-stimulating factor, erythropoietin, and thrombopoietin. A limiting dilution analysis of long-term culture-initiating cells (LTC-IC) showed that the CD34(+)IL-6R(-) cells contained a larger number of LTC-IC than did the CD34(+)IL-6R(+) cells. In a serum-free suspension of CD34(+)IL-6R(-) cells, the addition of sIL-6R to the combination of IL-6 and SCF dramatically increased the total and multipotential progenitors, whereas CD34(+)IL-6R(+) cells failed to do so under the same conditions. These results indicate that most of the erythroid, megakaryocytic, and primitive human hematopoietic progenitors are included in the IL-6R(-) populations, and the activation of gp130 on these progenitors can be achieved by a complex of IL-6-sIL-6R, but not by IL-6 alone. The present culture system using IL-6, sIL-6R, and SCF may provide a novel approach for ex vivo expansion of human primitive hematopoietic progenitors.