In vitro modulation of proliferation and phenotype of resting and mitogen-stimulated bovine mononuclear leukocytes by 1,25-dihydroxyvitamin D3.
Effects of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] on the proliferation and phenotype of normal bovine peripheral blood mononuclear leukocytes (MNL) were studied in vitro. Resting and pokeweed mitogen (PWM)-stimulated MNL cultures were supplemented with 1.0 nM of 1,25(OH)2D3 at the beginning of the culture period. Leukocytes were removed from 2-, 4-, 6-, 8-, 10-, 12-, and 14-day unsupplemented and 1.25(OH)2D3-supplemented cultures, and were counted and phenotyped using monoclonal antibodies to bovine leukocyte surface antigens. Cell numbers in resting MNL cultures decreased with time and were unaffected by 1,25(OH)2D3 supplementation. The progressive increase in cell numbers in PWM-stimulated MNL cultures was suppressed, but not abolished by 1,25(OH)2D3. Suppression was greatest in PWM-stimulated 6- to 12-day cultures. Cellular composition of resting MNL cultures was unaffected by 1,25(OH)2D3. Pokeweed mitogen-stimulated cultures supplemented with 1,25(OH)2D3 had fewer total T-cells and CD4+ T-cells at 6-14 days. In contrast, numbers of CD8+ T-cells were significantly higher in 1,25(OH)2D3-supplemented cultures at 6, 10, and 14 days. Effects of 1,25(OH)2D3 on CD4+ and CD8+ T-cell populations were manifested by a significant reduction in the CD4+:CD8+ T-cell ratios in 6- to 14-day cultures. Proliferation of IL-2 receptor+ and MHC class II antigen+ cells was also reduced in supplemented 6- to 14-day cultures, indicating events associated with PWM-induced MNL activation were suppressed by 1,25(OH)2D3. These findings indicate that 1,25(OH)2D3 modulates the proliferation and differentiation of bovine MNL in vitro. Our results also suggest that changes in plasma or tissue 1,25(OH)2D3 concentrations that occur during the peripartum period and in clinical cases of milk fever may regulate the bovine immune system in vivo.
Nonnecke
B J
BJ
United States Department of Agriculture, National Animal Disease Center, Ames, IA 50010-0070.
Franklin
S T
ST
Reinhardt
T A
TA
Horst
R L
RL
D000818:Animals; D000954:Antigens, Surface / Q000276:immunology; D016516:CD4-CD8 Ratio; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D002117:Calcitriol / Q000276:immunology*; D002417:Cattle; D002455:Cell Division; D002478:Cells, Cultured; D005260:Female; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D016130:Immunophenotyping; D007376:Interleukin-2 / Q000276:immunology; D007958:Leukocyte Count; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D008213:Lymphocyte Activation; D011043:Pokeweed Mitogens; D050378:T-Lymphocytes, Regulatory / Q000276:immunology
D000954:Antigens, Surface; D000949:Histocompatibility Antigens Class II; D007376:Interleukin-2; D011043:Pokeweed Mitogens; D002117:Calcitriol