Synthetic CpG-ODN rapidly enriches immune compartments in neonatal chicks to induce protective immunity against bacterial infections
Synthetic CpG-ODN rapidly enriches immune compartments in neonatal chicks to induce protective immunity against bacterial infections
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DOI：
10.1038/s41598-018-36588-6
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发表时间：
2019-01-23
影响因子：
4.6
通讯作者：
Gomis, Susantha
中科院分区：
文献类型：
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作者：
Gunawardana, Thushari;Ahmed, Khawaja Ashfaque;Gomis, Susantha
含有CpG基序的寡聚脱氧核苷酸（CpG-ODN）诱导针对细菌感染的先天免疫。尽管最近取得了进展，但CpG-ODN如何单独保护免受细菌感染仍然难以捉摸。在这里，我们报告的第一次，据我们所知，CpG-ODN编排抗微生物保护性免疫诱导快速富集的各种免疫区室鸡。在该研究中，用50 μ g CpG-ODN或盐水（类似于每组n = 90）注射18天大的含胚卵。在第一个实验中，CpG-ODN处理后四天，用大肠杆菌（E. coli）强毒株皮下攻击小鸡。coli），并监测死亡率8天。我们发现CpG-ODN处理的雏鸡具有显著的保护作用，并降低了临床评分。为了深入了解CpG-ODN诱导的保护机制，我们首先研究了CpG-ODN诱导的细胞因子表达动力学。在CpG-ODN接种后的10个时间点，从胚胎或鸡（每组n = 3-4）收集脾和肺。多重基因分析（白细胞介素（IL）-1、IL-4、IL-6、IL-10、IL-18、干扰素（IFN）-γ、IFN-α和脂多糖诱导的肿瘤坏死因子（LITAF））显示，与盐水对照相比，CpG-ODN处理后促炎细胞因子的表达显著更高。在我们的研究中，LITAF在脾和肺的细胞因子谱中脱颖而出，强调了其在CpG-ODN诱导的保护中的作用。实验三检测CpG-ODN对脾、肺和胸腺免疫细胞群的影响。在CpG-ODN给药后24、48和72小时（仅在72小时收集胸腺）进行流式细胞术分析，以检测CD 4（+）和CD 8（+）T细胞亚群、单核细胞/巨噬细胞群体及其成熟标志物（CD 40和CD 86）表达的变化。流式细胞仪检测结果显示，CpG-ODN处理的鸡胚和鸡胚脾和肺中巨噬细胞、CD 4+和CD 8 + T细胞亚群显著富集。脾和肺中的巨噬细胞显示出上调的CD 40，但不是CD 86，而胸腺细胞显示出显着高的CD 4和CD 8的表达。总之，本研究表明CpG-ODN对新生雏鸡具有抗大肠杆菌的保护作用。大肠杆菌感染不仅通过引发细胞因子反应和刺激免疫细胞，而且还通过丰富脾和肺中的免疫小生境。
Oligodeoxynucleotides containing CpG motifs (CpG-ODN) induce innate immunity against bacterial infections. Despite recent advances, how CpG-ODN alone protects against bacterial infections remained elusive. Here, we report for the first time, to our knowledge, that CpG-ODN orchestrates anti-microbial protective immunity by inducing a rapid enrichment of various immune compartments in chickens. In this study, eighteen-day-old embryonated eggs were injected with either 50 mu g of CpG-ODN or saline (similar to n = 90 per group). In the first experiment, four days after CpG-ODN treatment, chicks were challenged subcutaneously with a virulent strain of Escherichia coli (E. coli) and mortality was monitored for 8 days. We found significant protection, and reduced clinical scores in CpG-ODN treated chicks. To gain insights into mechanisms of protection induced by CpG-ODN, first we investigated cytokine expression kinetics elicited by CpG-ODN. The spleen and lung were collected from embryos or chicks (n = 3-4 per group) at 10 time points post-CpG-ODN inoculation. Multiplex gene analysis (interleukin (IL)-1, IL-4, IL-6, IL-10, IL-18, interferon (IFN)-gamma, IFN-alpha, and lipopolysaccharide induced tumor necrosis factor (LITAF), revealed a significantly higher expression of pro-inflammatory cytokines following CpG-ODN treatment compared to the saline controls. In our study, LITAF stands out in the cytokine profiles of spleen and lungs, underscoring its role in CpG-ODN-induced protection. The third experiment was designed to examine the effects of CpG-ODN on immune cell populations in spleen, lungs, and thymus. Flow cytometry analysis was conducted at 24, 48 and 72 hrs (thymus only collected at 72 hr) after CpG-ODN administration to examine the changes in CD4(+) and CD8(+) T-cell subsets, monocyte/macrophage cell populations and their expression of maturation markers (CD40 and CD86). Flow cytometry data indicated a significant enrichment of macrophages, CD4(+) and CD8(+) T-cell subsets in both spleen and lungs of CpG-ODN treated embryos and chicks. Macrophages in spleen and lungs showed an upregulation of CD40 but not CD86, whereas thymocytes revealed significantly high CD4 and CD8 expression. Overall, the present study has demonstrated that CpG-ODN provides protection in neonatal chicks against E. coli infection not only by eliciting cytokine responses and stimulating immune cells but also through enriching immunological niches in spleen and lungs.