Table_6_Identifying transcriptomic profiles in ovine spleen after repetitive vaccination.xlsx

Aluminum hydroxide has long been employed as a vaccine adjuvant for its safety profile, although its precise mechanism of action remains elusive. In this study, we investigated the transcriptomic responses in sheep spleen following repetitive vaccination with aluminum adjuvanted vaccines and aluminum hydroxide alone. Notably, this work represents the first exploration of the sheep spleen transcriptome in such conditions. Animals were splitted in 3 treatment groups: vaccine group, adjuvant alone group and control group. A total of 18 high-depth RNA-seq libraries were sequenced, resulting in a rich dataset which also allowed isoform-level analysis. The comparisons between vaccine-treated and control groups (V vs C) as well as between vaccine-treated and adjuvant-alone groups (V vs A) revealed significant alterations in gene expression profiles, including protein coding genes and long non-coding RNAs. Among the differentially expressed genes, many were associated with processes such as endoplasmic reticulum (ER) stress, immune response and cell cycle. The analysis of co-expression modules further indicated a correlation between vaccine treatment and genes related to ER stress and unfolded protein response. Surprisingly, adjuvant-alone treatment had little impact on the spleen transcriptome. Additionally, the role of alternative splicing in the immune response was explored. We identified isoform switches in genes associated with immune regulation and inflammation, potentially influencing protein function. In conclusion, this study provides valuable insights into the transcriptomic changes in sheep spleen following vaccination with aluminum adjuvanted vaccines and aluminum hydroxide alone. These findings shed light on the molecular mechanisms underlying vaccine-induced immune responses and emphasize the significance of antigenic components in aluminum adjuvant mechanism of action. Furthermore, the analysis of alternative splicing revealed an additional layer of complexity in the immune response to vaccination in a livestock species.

氢氧化铝（aluminum hydroxide）凭借优异的安全性，长期被用作疫苗佐剂，但其确切的作用机制仍未明确。本研究针对重复接种铝佐剂疫苗及单纯氢氧化铝佐剂后绵羊脾脏的转录组响应展开探究，值得注意的是，本研究首次在该类条件下对绵羊脾脏转录组进行相关分析。实验动物被分为3个处理组：疫苗组、单纯佐剂组与对照组。最终共完成18个高深度RNA测序（RNA-seq）文库的测序，获得了高质量的数据集，该数据集同时支持异构体水平的分析。对疫苗处理组与对照组（V vs C）、疫苗处理组与单纯佐剂组（V vs A）的比较分析显示，两组的基因表达谱均发生显著改变，涉及蛋白编码基因与长链非编码RNA（long non-coding RNAs）。在差异表达基因中，大量基因与内质网（ER）应激、免疫应答及细胞周期等生物学过程相关。共表达模块分析进一步揭示，疫苗处理与内质网应激及未折叠蛋白反应相关基因存在显著关联。令人意外的是，单纯佐剂处理对绵羊脾脏转录组几乎无影响。此外，本研究还探究了可变剪接（alternative splicing）在免疫应答中的作用，在免疫调控与炎症相关基因中发现了异构体转换现象，该现象可能会影响蛋白质的功能。综上，本研究为重复接种铝佐剂疫苗及单纯氢氧化铝佐剂后绵羊脾脏的转录组变化提供了重要见解，其结果阐明了疫苗诱导免疫应答的潜在分子机制，并凸显了抗原成分在氢氧化铝佐剂作用机制中的重要性。此外，可变剪接分析还揭示了家畜疫苗接种免疫应答中另一层复杂的调控机制。