INRA, UMR1313 Génétique Animale et Biologie Intégrative, F-78350 Jouy-en-Josas, France.
BMC Genomics. 2013 Dec 17;14:894. doi: 10.1186/1471-2164-14-894.
Immune traits (ITs) are potentially relevant criteria to characterize an individual's immune response. Our aim was to investigate whether the peripheral blood transcriptome can provide a significant and comprehensive view of IT variations in pig.
Sixty-day-old Large White pigs classified as extreme for in vitro production of IL2, IL10, IFNγ and TNFα, phagocytosis activity, in vivo CD4⁻/CD8⁺ or TCRγδ + cell counts, and anti-Mycoplasma antibody levels were chosen to perform a blood transcriptome analysis with a porcine generic array enriched with immunity-related genes. Differentially expressed (DE) genes for in vitro production of IL2 and IL10, phagocytosis activity and CD4⁻/CD8⁺ cell counts were identified. Gene set enrichment analysis revealed a significant over-representation of immune response functions. To validate the microarray-based results, a subset of DE genes was confirmed by RT-qPCR. An independent set of 74 animals was used to validate the covariation between gene expression levels and ITs. Five potential gene biomarkers were found for prediction of IL2 (RALGDS), phagocytosis (ALOX12) or CD4⁻/CD8⁺ cell count (GNLY, KLRG1 and CX3CR1). On average, these biomarkers performed with a sensitivity of 79% and a specificity of 86%.
Our results confirmed that gene expression profiling in blood represents a relevant molecular phenotype to refine ITs in pig and to identify potential biomarkers that can provide new insights into immune response analysis.
免疫特征(ITs)是描述个体免疫反应的潜在相关标准。我们的目的是研究外周血转录组是否可以提供猪 IT 变化的显著而全面的观点。
选择了在体外产生 IL2、IL10、IFNγ 和 TNFα、吞噬活性、体内 CD4⁻/CD8⁺或 TCRγδ+细胞计数以及抗支原体抗体水平方面表现出极端的 60 日龄长白猪,进行了血液转录组分析,使用了富含免疫相关基因的猪通用阵列。鉴定了与体外产生 IL2 和 IL10、吞噬活性和 CD4⁻/CD8⁺细胞计数相关的差异表达(DE)基因。基因集富集分析显示免疫反应功能显著过表达。为了验证微阵列结果,通过 RT-qPCR 确认了 DE 基因的子集。使用 74 只动物的独立集来验证基因表达水平与 IT 之间的共变关系。发现了 5 个预测 IL2(RALGDS)、吞噬作用(ALOX12)或 CD4⁻/CD8⁺细胞计数(GNLY、KLRG1 和 CX3CR1)的潜在基因生物标志物。平均而言,这些生物标志物的灵敏度为 79%,特异性为 86%。
我们的结果证实,血液中的基因表达谱代表了一种相关的分子表型,可以细化猪的 ITs,并确定潜在的生物标志物,为免疫反应分析提供新的见解。