Anti-inflammatory effects of retinoids and carotenoid derivatives on caspase-3-dependent apoptosis and efferocytosis of bovine neutrophils

Objective-To evaluate immunomodulatory properties of all-trans retinoic acid and a fully oxidized beta-carotene dietary product in calves with Mannheimia haemolytica-induced pneumonia. Animals-Twenty-five 6-to 10-week-old male Holstein calves for experimental inoculations and three 8-to 30-week-old Angus heifers for blood donations. Procedures-In vitro, neutrophils and monocyte-derived macrophages isolated from blood of healthy Angus heifers were treated with all-trans retinoic acid (1 mu M) or fully oxidized beta-carotene (8.3 mu g/mL) for various times and assessed for markers of cellular death, antimicrobial function, and production of proinflammatory leukotriene B-4. Following 28 days of dietary supplementation with fully oxidized beta-carotene, Holstein calves were experimentally inoculated with M haemolytica. Bronchoalveolar lavage fluid was collected at 3 and 24 hours after challenge inoculation and analyzed for markers of apoptosis. Results-In vitro, all-trans retinoic acid and fully oxidized beta-carotene induced cell-selective, caspase-3-dependent apoptosis in neutrophils, which subsequently enhanced efferocytosis in macrophages. Conversely, neither treatment altered phorbol 12-myristate 13-acetate-induced oxidative burst, phagocytosis of nonopsonized zymosan (complement or antibody independent), or M haemolytica-induced leukotriene B-4 production in bovine neutrophils. In vivo, fully oxidized beta-carotene enhanced leukocyte apoptosis in bronchoalveolar lavage fluid as well as subsequent efferocytosis by macrophages without altering numbers of circulating leukocytes. Conclusions and Clinical Relevance-Neutrophil apoptosis and subsequent efferocytosis by macrophages are key mechanisms in the resolution of inflammation. Findings for the present study indicated that all-trans retinoic acid and fully oxidized beta-carotene could be novel nutraceutical strategies that may confer anti-inflammatory benefits for cattle with respiratory tract disease.