Animal & Bioscience Research Department, Animal & Grassland Research and Innovation Centre, Teagasc, Grange, Co Meath, Ireland.
School of Biochemistry and Immunology, Trinity College Dublin, Dublin 2, Ireland.
Sci Rep. 2023 Jul 5;13(1):10846. doi: 10.1038/s41598-023-37427-z.
Vitamin D deficiency at birth, followed by prolonged insufficiency in early life may predispose bovine calves to infection and disease. However, the effects of vitamin D levels on innate immunity are unclear due to the lack of long-term supplementation trials in vivo and reliable approaches for reproducibly assessing immune function. Here, a standardized whole blood immunophenotyping assay was used to compare innate immune responses to infection relevant ligands (LPS, Pam3CSK4 and R848) between Holstein-Friesian calves supplemented with vitamin D (n = 12) from birth until 7 months of age and control calves (n = 10) raised on an industry standard diet. Transcriptomic analysis in unstimulated whole blood cells revealed increased expression of type I interferons and chemokines in vitamin D supplemented calves, while IL-1 and inflammasome gene expression was decreased. In response to stimulation with the bacterial ligand LPS, supplemented calves had significantly increased expression of CASP1, CX3CR1, CAT, whereas STAT1 was decreased. Stimulation with the bacterial ligand Pam3CSK4 revealed increased expression of IL1A, IL1B and CAT genes; and decreased C5AR1 expression. In response to the viral ligand R848, STAT1 and S100A8 expression was significantly decreased. An increased IL-1 and inflammasome gene expression signature in vitamin D supplemented calves in response to LPS and Pam3CSK4 was also found, with ELISA confirming increased IL-1β protein production. In contrast, a decreased chemokine gene expression signature was found in response to R848 in supplemented animals, with decreased IL-8 protein expression exhibited in response to all PAMPs also found. These results demonstrated expression of several cytokine, chemokine and inflammasome genes were impacted by vitamin D supplementation in the first 7 months of life, with IL-8 expression particularly responsive to vitamin D. Overall, vitamin D supplementation induced differential innate immune responses of blood immune cells that could have important implications for disease susceptibility in cattle.
出生时维生素 D 缺乏，随后在生命早期长期不足，可能使牛犊易感染和患病。然而，由于缺乏长期的体内补充试验和可靠的方法来重现性评估免疫功能，维生素 D 水平对先天免疫的影响尚不清楚。在这里，使用标准化的全血免疫表型分析方法比较了从出生到 7 个月大补充维生素 D（n = 12）的荷斯坦-弗里森牛犊与在行业标准饮食下饲养的对照牛犊（n = 10）对感染相关配体（LPS、Pam3CSK4 和 R848）的先天免疫反应。未刺激的全血细胞转录组分析显示，补充维生素 D 的牛犊中 I 型干扰素和趋化因子的表达增加，而 IL-1 和炎症小体基因的表达减少。在对细菌配体 LPS 的刺激下，补充组的 CASP1、CX3CR1 和 CAT 表达显著增加，而 STAT1 减少。在对细菌配体 Pam3CSK4 的刺激下，IL1A、IL1B 和 CAT 基因的表达增加，C5AR1 表达减少。在对病毒配体 R848 的反应中，STAT1 和 S100A8 的表达显著降低。还发现，在 LPS 和 Pam3CSK4 刺激下，补充维生素 D 的牛犊中出现了增加的 IL-1 和炎症小体基因表达特征，ELISA 证实了 IL-1β 蛋白的产生增加。相反，在补充动物中发现了对 R848 的趋化因子基因表达特征降低，并且还发现所有 PAMP 反应时 IL-8 蛋白表达降低。这些结果表明，在生命的前 7 个月中，维生素 D 补充会影响几种细胞因子、趋化因子和炎症小体基因的表达，其中 IL-8 表达对维生素 D 特别敏感。总的来说，维生素 D 补充诱导了血液免疫细胞的先天免疫反应的差异，这可能对牛的疾病易感性有重要影响。
