Regulation of Mycobacterium-Specific Mononuclear Cell Responses by 25-Hydroxyvitamin D3

The active vitamin D metabolite, 1,25-dihydroxyvitamin D-3 (1,25(OH)(2)D-3), has been shown to be an important regulator of innate and adaptive immune function. In addition, synthesis of 1,25(OH)(2)D-3 from 25-hydroxyvitamin D-3 (25(OH)D-3) by the enzyme 1 alpha-hydroxylase in monocytes upon activation by TLR signaling has been found to regulate innate immune responses of monocytes in an intracrine fashion. In this study we wanted to determine what cells expressed 1 alpha-hydroxylase in stimulated peripheral blood mononuclear cell (PBMC) cultures and if conversion of 25(OH)D-3 to 1,25(OH)(2)D-3 in PBMC cultures regulated antigen-specific immune responses. Initially, we found that stimulation of PBMCs from animals vaccinated with Mycobacterium bovis (M. bovis) BCG with purified protein derivative of M. bovis (M. bovis PPD) induced 1 alpha-hydroxylase gene expression and that treatment with a physiological concentration of 25(OH)D-3 down-regulated IFN-gamma and IL-17F gene expression. Next, we stimulated PBMCs from M. bovis BCG-vaccinated and non-vaccinated cattle with M. bovis PPD and sorted them by FACS according to surface markers for monocytes/macrophages (CD14), B cells (IgM), and T cells (CD3). Sorting the PBMCs revealed that 1 alpha-hydroxylase expression was induced in the monocytes and B cells, but not in the T cells. Furthermore, treatment of stimulated PBMCs with 25(OH) D-3 down-regulated antigen-specific IFN-gamma and IL-17F responses in the T cells, even though 1 alpha-hydroxylase expression was not induced in the T cells. Based on evidence of no T cell 1a-hydroxylase we hypothesize that activated monocytes and B cells synthesize 1,25(OH)(2)D-3 and that 1,25(OH)(2)D-3 down-regulates antigen-specific expression oIFN-gamma and IL-17F in T cells in a paracrine fashion.