Universidad Nacional de Colombia, sede Bogotá. Facultad de Medicina Veterinaria y de Zootecnia. Departamento de Salud Animal. Centro de Investigación en Inmunología e Infectología Veterinaria (CI(3)V), Bogotá CP 11001, Colombia.
Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph N1G 2W1, ON, Canada.
Poult Sci. 2020 Sep;99(9):4265-4277. doi: 10.1016/j.psj.2020.06.006. Epub 2020 Jun 23.
It has been demonstrated that vitamin D (Vit D) included in diets offers a beneficial effect by improving innate immune responses in chickens. However, its mechanisms of action and the effect on immunosuppressive pathogens, such as infectious bursal disease virus, are not yet known. In the present study, we have studied the immunomodulatory effect of Vit D on the innate immune response in 3 cell lines: fibroblast cells (DF-1), macrophages (HD11), and B cells (DT-40) infected with IBDV (intermediate vaccine) at 2 multiplicity of infections (MOI) (1 and 0.1). Genes associated with innate immune responses (TLR-3, TLR-21, MDA-5, MyD88, TRIF, IRF-7, INF-α, INF-β, PKR, OAS, viperin, IL-1β, IL-6, and IL-12) were evaluated at different time points (3, 6, 12, 24, and 36 h after infection, h.p.i). Virus production reached a maximum at 24 h.p.i., which was significantly (P < 0.05) higher in DF-1 cells, followed by HD-11 and DT-40 cells. Mainly in HD-11 cells, there was a significant (P < 0.05) effect of Vit D supplementation on receptors TLR-3, TLR-21, and MDA-5 after 12 h.p.i, independent of MOI. DT-40 cells showed the highest antiviral activity, with a significant (P < 0.05) effect on IRF-7, IFN-β, OAS, and PKR gene expression, where expression of IRF-7 and IFN-β correlated positively with Vit D supplementation, while OAS and PKR were independent of Vit D. Proinflammatory cytokines were significantly (P < 0.05) upregulated and found to be Vit D and MOI dependent. In conclusion, this study demonstrated the capacity of IBDV to trigger a strong innate immune response in chicken cells and contributes to the understanding of the activation pathways of innate immunity induced by IBDV and further shows the benefitial effect of Vit D supplementation as an immunomodulator.
已经证明,膳食中的维生素 D(Vit D)通过改善鸡的先天免疫反应提供有益效果。然而,其作用机制以及对免疫抑制性病原体(如传染性法氏囊病病毒)的影响尚不清楚。在本研究中,我们研究了 Vit D 对感染 IBDV(中等疫苗)的 3 种细胞系(成纤维细胞(DF-1)、巨噬细胞(HD11)和 B 细胞(DT-40))先天免疫反应的免疫调节作用,感染复数(MOI)为 2(1 和 0.1)。在感染后不同时间点(感染后 3、6、12、24 和 36 小时,h.p.i.)评估了与先天免疫反应相关的基因(TLR-3、TLR-21、MDA-5、MyD88、TRIF、IRF-7、INF-α、INF-β、PKR、OAS、viperin、IL-1β、IL-6 和 IL-12)。病毒产量在 24 h.p.i.达到最大值,DF-1 细胞中的病毒产量明显更高(P < 0.05),其次是 HD-11 和 DT-40 细胞。主要在 HD-11 细胞中,Vit D 补充剂在感染后 12 h.p.i.时对 TLR-3、TLR-21 和 MDA-5 受体有显著(P < 0.05)的影响,而与 MOI 无关。DT-40 细胞表现出最高的抗病毒活性,IRF-7、IFN-β、OAS 和 PKR 基因表达有显著(P < 0.05)的影响,IRF-7 和 IFN-β的表达与 Vit D 补充呈正相关,而 OAS 和 PKR 与 Vit D 无关。促炎细胞因子的表达明显上调(P < 0.05),且与 Vit D 和 MOI 有关。综上所述,本研究证明了 IBDV 能够在鸡细胞中引发强烈的先天免疫反应,并有助于了解 IBDV 诱导的先天免疫激活途径,进一步表明 Vit D 补充作为免疫调节剂的有益效果。