Exogenous Vitamin D3 Modulates Response of Bovine Macrophages to Mycobacterium avium subsp. paratuberculosis Infection and Is Dependent Upon Stage of Johne's Disease

Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of ruminant enteritis, targets intestinal macrophages. During infection, macrophages contribute to mucosal inflammation and development of granulomas in the small intestine which worsens as disease progression occurs. Vitamin D-3 is an immunomodulatory steroid hormone with beneficial roles in host-pathogen interactions. Few studies have investigated immunologic roles of 25-hydroxyvitamin D-3 (25(OH)D-3) and 1,25-dihydroxyvitamin D-3 (1,25(OH)(2)D-3) in cattle, particularly cattle infected with MAP. This study examined the effects of exogenous vitamin D-3 on immune responses of monocyte derived macrophages (MDMs) isolated from dairy cattle naturally infected with MAP. MDMs were pre-treated with +/- 100 ng/ml 25(OH)D-3 or +/- 4 ng/ml 1,25(OH)(2)D-3, then incubated 24 hrs with live MAP in the presence of their respective pre-treatment concentrations. Following treatment with either vitamin D-3 analog, phagocytosis of MAP by MDMs was significantly greater in clinically infected animals, with a greater amount of live and dead bacteria. Clinical cows had significantly less CD40 surface expression on MDMs compared to subclinical cows and noninfected controls. 1,25(OH)(2)D-3 also significantly increased nitrite production in MAP infected cows. 1,25(OH)(2)D-3 treatment played a key role in upregulating secretion of pro-inflammatory cytokines IL-1 beta and IL-12 while downregulating IL-10, IL-6, and IFN-gamma. 1,25(OH)(2)D-3 also negatively regulated transcripts of CYP24A1, CYP27B1, DEFB7, NOS2, and IL10. Results from this study demonstrate that vitamin D-3 compounds, but mainly 1,25(OH)(2)D-3, modulate both pro- and anti-inflammatory immune responses in dairy cattle infected with MAP, impacting the bacterial viability within the macrophage.