Department of Agricultural, Food and Nutritional Science, University of Alberta, Canada.
Department of Agricultural, Food and Nutritional Science, University of Alberta, Canada; Livestock Gentec, University of Alberta, Canada.
Vaccine. 2019 Mar 22;37(13):1743-1755. doi: 10.1016/j.vaccine.2019.02.016. Epub 2019 Feb 23.
We investigated gene expression patterns in whole blood and fecal microbiota profile as potential predictors of immune response to vaccination, using healthy M. hyopneumoniae infection free piglets (n = 120). Eighty piglets received a dose of prophylactic antibiotics during the first two days of life, whereas the remaining 40 did not. Blood samples for RNA-Seq analysis were collected on experimental Day 0 (D0; 28 days of age) just prior to vaccination, D2, and D6 post-vaccination. A booster vaccine was given at D24. Fecal samples for microbial 16SrRNA sequencing were collected at 7 days of age, and at D0 and D35 post-vaccination. Pigs were ranked based on the levels of M. hyopneumoniae-specific antibodies in serum samples collected at D35, and groups of 'high' (HR) and 'low' (LR) responder pigs (n = 15 each) were selected. Prophylactic antibiotics did not influence antibody titer levels and differential expression analysis did not reveal differences between HR and LR at any time-point (FDR > 0.05); however, based on functional annotation with Ingenuity Pathway Analysis, D2 post-vaccination, HR pigs were enriched for biological terms relating to increased activation of immune cells. In contrast, the immune activation decreased in HR, 6 days post-vaccination. No significant differences were observed prior to vaccination (D0). Two days post-vaccination, multivariate analysis revealed that ADAM8, PROSER3, B4GALNT1, MAP7D1, SPP1, HTRA4, and ENO3 genes were the most promising potential biomarkers. At D0, OTUs annotated to Prevotella, CF21, Bacteroidales and S24-7 were more abundant in HR, whereas Fibrobacter, Paraprevotella, Anaerovibrio, [Prevotella], YRC22, and Helicobacter positively correlated with the antibody titer as well as MYL1, SPP1, and ENO3 genes. Our study integrates gene differential expression and gut microbiota to predict vaccine response in pigs. The results indicate that post-vaccination gene-expression and early-life gut microbiota profile could potentially predict vaccine response in pigs, and inform a direction for future research.
我们研究了全血中的基因表达模式和粪便微生物群的特征，作为对疫苗免疫反应的潜在预测因子，使用了健康的支原体感染自由仔猪（n=120）。80 头仔猪在生命的头两天接受了预防性抗生素治疗，而其余 40 头则没有。用于 RNA-Seq 分析的血液样本于实验第 0 天（D0；28 天龄）在接种疫苗前、接种后第 2 天（D2）和第 6 天（D6）采集。在 D24 时给予加强疫苗。粪便样本用于微生物 16SrRNA 测序，于 7 天龄时采集，以及接种疫苗后第 0 天和第 35 天采集。根据第 35 天采集的血清样本中支原体特异性抗体的水平对猪进行排序，并选择了“高”（HR）和“低”（LR）应答猪组（每组 15 头）。预防性抗生素不会影响抗体滴度水平，差异表达分析在任何时间点均未显示 HR 和 LR 之间的差异（FDR＞0.05）；然而，基于 Ingenuity Pathway Analysis 的功能注释，接种疫苗后第 2 天，HR 猪富集了与免疫细胞激活增加相关的生物学术语。相比之下，接种疫苗后 6 天，HR 中的免疫激活降低。接种疫苗前（D0）未观察到显著差异。接种疫苗后第 2 天，多变量分析显示 ADAM8、PROSER3、B4GALNT1、MAP7D1、SPP1、HTRA4 和 ENO3 基因是最有前途的潜在生物标志物。在 D0，注释为 Prevotella、CF21、Bacteroidales 和 S24-7 的 OTUs 在 HR 中更为丰富，而 Fibrobacter、Paraprevotella、Anaerovibrio、[Prevotella]、YRC22 和 Helicobacter 与抗体滴度以及 MYL1、SPP1 和 ENO3 基因呈正相关。我们的研究将基因差异表达与肠道微生物群相结合，以预测猪的疫苗反应。结果表明，接种疫苗后的基因表达和早期生活肠道微生物群特征可能潜在地预测猪的疫苗反应，并为未来的研究提供方向。
