Influence of vitamin D3 infusion and dietary calcium on secretion of interleukin 1, interleukin 6, and tumor necrosis factor in mice infected with Mycobacterium paratuberculosis

Abstract

Objective - To study the effects of 1,25(OH)2D3 and calcium (Ca) on splenocyte cytokine secretion during Mycobacterium paratuberculosis infection. Design - Mice were assigned to the following treatments: 1-noninfected, 2-infected, 3-noninfected/1,25-(OH)2D3, 4-infected/1,25(OH)2D3, and 5 - infected/low-Ca diet (0.15%). Animals - Male beige mice averaging 6 weeks of age and 20 g in body weight. Procedure - After acclimation to their diets, mice in treatments 2, 4, and 5 were inoculated IV with 108 colony-forming units of M paratuberculosis. At 1, 6, and 12 months after infection, mice in treatment groups 3 and 4 had miniosmotic pumps implanted subcutaneously that delivered 13 ng of 1,25(OH)2D3/day for 14 days. Treatment 5 was included as a control for comparison with treatment 4 because low dietary Ca should increase endogenous 1,25(OH)2D3 values. Splenocytes were isolated from mice at 1, 6, and 12 months and stimulated in vitro with medium alone (nonstimulated), lipopolysaccharide (LPS), concanavalin A, and M paratuberculosis whole-cell sonicate (MpS). Results - Production of interleukin 6 after stimulation with LPS, concanavalin A, or MpS was higher (P < 0.05) for splenocytes isolated from mice fed the low-Ca diet, compared with control infected mice 1 and 6 months after infection. Interleukin 1 and tumor necrosis factor activities were increased (P < 0.05) in splenocytes cultured with LPS and MpS after isolation from mice of the low-Ca group. Mice infused with 1,25(OH)2D3 had higher (P < 0.05) interleukin 1 secretion after stimulation of splenocytes with LPS and higher (P < 0.05) tumor necrosis factor production after incubation with MpS. Conclusion - 1,25(OH)2D3 and low dietary Ca increase cytokine secretion in mice infected with M paratuberculosis.