Recombinant human (rh) stem cell factor and rhIL-4 stimulate differentiation and proliferation of CD3 + cells from umbilical cord blood and CD3 + cells enhance FcεR1 expression on fetal liver-derived mast cells in the presence of rhIL-4

Abstract

We previously reported that rhIL-4 induced apoptosis and rhIL-6 mediated protection of human mast cells derived from cord blood mononuclear cells. Based on the result, we attempted to obtain the phenotypes and differentiation of CD3 + cells from cord blood by investigating their cell surface markers in the presence of rhSCF plus rhIL-4. The effect of co-cultured CD3 + cells on fetal liver mast cells (FLMCs) was also determined. Phenotypes from cord blood-derived cells were analyzed by flow cytometry and cell numbers were determined. Fetal liver mast cells were cultured with cord blood-derived cells (mainly CD3 +) in the presence of rhSCF and/or rhIL-4 and were analyzed to determine cell number and expression of Kit + and FcεR1. The percentage of CD3 + cells from cord blood-derived cells on day 0 was about 41 ± 13.5%, following monocytes and granulocytes. CD3 + cells increased in number (1.5-fold) and purity (90%), whereas other cell types did not survive. More than 60% of CD3 + cells from cord blood at day 0 were CD4 −CD8 −. These double-negative cells dramatically decreased by 1 week of culture, while CD4 +CD8 + cells increased in number and purity through 3 weeks of culture, and then decreased as greater numbers of single-positive T cells emerged. We also found that FcεR expression on FLMC increased in the presence of rhIL-4, but was not affected by the T cells that developed from cord blood mononuclear cells. The results indicate that IL-4, a Th2 type cytokine, together with rhSCF, can induce T cell proliferations, differentiation, and maturation from cord blood progenitor cells.