Induction of chemokine mRNA in bone marrow stromal cells: modulation by TGF-beta 1 and IL-4

Bone marrow stromal cells produce cytokines that are essential for the proliferation and differentiation of hematopoietic stem and progenitor cells. Thus, regulation of cytokine production by bone marrow accessory cells is a critical aspect of stromal cell regulation of hematopoiesis. We have investigated the effect of two cytokines that have been demonstrated to modulate factor production by non-marrow accessory cells (i.e., transforming growth factor-beta 1 [TGF-beta 1] and interleukin-4 [IL-4]) on the induced expression of cytokine mRNA in a bone marrow-derived, cloned, murine stromal cell line +/+/-.LDA11. We showed that +/+/-.LDA11 cells can be induced with lipopolysaccharide (LPS), IL-1 alpha, or interferon-gamma (IFN-gamma) to express mRNA for monocyte chemoattractant protein-1 (MCP-1/JE), IFN-inducible protein-10 (IP-10), stem cell factor (SCF), and macrophage colony-stimulating factor (M-CSF) but not for IL-1 alpha, IL-3, or tumor necrosis factor-alpha (TNF-alpha). The expression of MCP-1/JE and IP-10 mRNA by these inducers was potentiated by TGF-beta 1 and IL-4. The augmentation by TGF-beta 1 of both mRNAs induced with IL-1 alpha was maximum when applied to the cells concurrently with the inducer; the IFN-gamma-induced expression of mRNAs was augmented even if the addition of TGF-beta 1 was delayed. Similarly, IL-4 potentiation of both mRNAs by either inducer progressively increased as the time between exposure to the inducer and exposure to IL-4 increased. Neither modulator altered the time course of mRNA expression by either inducer. TGF-beta 1- and IL-4-mediated augmentation of MCP-1/JE mRNA by IL-1 alpha or IFN-gamma was partially reversed by cycloheximide (CHX), whereas potentiation of IP-10 by either modulator remained unaffected. Increase in the stability of mRNA transcripts by TGF-beta 1 or IL-4 does not appear to play a role in the enhanced accumulation of mRNA in the presence of the modulators. These findings support a role for TGF-beta 1 and IL-4 as critical regulatory molecules in production of MCP-1 and IP-10 chemokines by stromal cells.