Cytokine and growth factor gene expression by bone marrow stroma of mice with damaged hematopoiesis and during regeneration

A study of gene expression of GM-CSF, IL-3, c-kit ligand, and IL-1 by bone marrow stromal cells of mice who were treated with an LD50 cytosine arabinoside (Ara-C) dose was carried out. It was shown previously that this dose causes extensive hematopoietic damage which is followed by regeneration in surviving mice. Using PCR, we demonstrated GM-CSF and IL-1 expression by adherent cells taken from marrow fibroblast layers following Ara-C-induced hematopoietic damage and during marrow regeneration, while expression in control layers was not detected. The IL-3 gene was not expressed either by layers of Ara-C-treated mice or by controls, probably due to the absence of T-lymphocytes in 2-4 week old cultures. The c-kit ligand gene was expressed by layers during marrow regeneration and by control layers, but was absent during the stage of hematopoietic damage. In parallel, in vitro cytokine production was evaluated. While IL-3 and GM-CSF were not present in the conditioned medium of marrow fibroblast layers either from Ara-C-treated mice or controls, IL-1 was found in low concentrations in cultures from Ara-C-treated animals. We conclude that GM-CSF, c-kit ligand and IL-1 have important roles in sustaining marrow regeneration following extensive damage. The role of IL-3 in marrow regeneration cannot be assessed in fibroblast layers of 2-4 week incubation where T-lymphocytes are generally absent.