INRA, UMR 1313 de Génétique Animale et Biologie Intégrative, Domaine de Vilvert, 78350 Jouy-en-Josas, France.
BMC Genomics. 2010 May 11;11:292. doi: 10.1186/1471-2164-11-292.
Designing sustainable animal production systems that better balance productivity and resistance to disease is a major concern. In order to address questions related to immunity and resistance to disease in pig, it is necessary to increase knowledge on its immune system and to produce efficient tools dedicated to this species.
A long-oligonucleotide-based chip referred to as SLA-RI/NRSP8-13K was produced by combining a generic set with a newly designed SLA-RI set that targets all annotated loci of the pig major histocompatibility complex (MHC) region (SLA complex) in both orientations as well as immunity genes outside the SLA complex. The chip was used to study the immune response of pigs following stimulation of porcine peripheral blood mononuclear cells (PBMCs) with lipopolysaccharide (LPS) or a mixture of phorbol myristate acetate (PMA) and ionomycin for 24 hours. Transcriptome analysis revealed that ten times more genes were differentially expressed after PMA/ionomycin stimulation than after LPS stimulation. LPS stimulation induced a general inflammation response with over-expression of SAA1, pro-inflammatory chemokines IL8, CCL2, CXCL5, CXCL3, CXCL2 and CCL8 as well as genes related to oxidative processes (SOD2) and calcium pathways (S100A9 and S100A12). PMA/ionomycin stimulation induced a stronger up-regulation of T cell activation than of B cell activation with dominance toward a Th1 response, including IL2, CD69 and TNFRSF9 (tumor necrosis factor receptor superfamily, member 9) genes. In addition, a very intense repression of THBS1 (thrombospondin 1) was observed. Repression of MHC class I genes was observed after PMA/ionomycin stimulation despite an up-regulation of the gene cascade involved in peptide processing. Repression of MHC class II genes was observed after both stimulations. Our results provide preliminary data suggesting that antisense transcripts mapping to the SLA complex may have a role during immune response.
The SLA-RI/NRSP8-13K chip was found to accurately decipher two distinct immune response activations of PBMCs indicating that it constitutes a valuable tool to further study immunity and resistance to disease in pig. The transcriptome analysis revealed specific and common features of the immune responses depending on the stimulation agent that increase knowledge on pig immunity.
设计能够更好地平衡生产力和疾病抵抗力的可持续动物生产系统是一个主要关注点。为了解决与猪的免疫和疾病抵抗力相关的问题，有必要增加对其免疫系统的了解，并生产专门针对该物种的有效工具。
通过将通用集与新设计的 SLA-RI 集相结合，制作了一种基于长寡核苷酸的芯片，该集针对猪主要组织相容性复合体（MHC）区域（SLA 复合体）的所有注释基因以及 SLA 复合体之外的免疫基因，以两种方向进行靶向。该芯片用于研究猪外周血单核细胞（PBMC）在脂多糖（LPS）或佛波醇肉豆蔻酸酯（PMA）和离子霉素混合物刺激 24 小时后的免疫反应。转录组分析显示，PMA/离子霉素刺激后差异表达的基因是 LPS 刺激后的 10 倍。LPS 刺激引起一般炎症反应，过表达 SAA1、促炎趋化因子 IL8、CCL2、CXCL5、CXCL3、CXCL2 和 CCL8 以及与氧化过程（SOD2）和钙途径（S100A9 和 S100A12）相关的基因。PMA/离子霉素刺激引起的 T 细胞激活上调强于 B 细胞激活，表现为 Th1 反应优势，包括 IL2、CD69 和 TNFRSF9（肿瘤坏死因子受体超家族，成员 9）基因。此外，还观察到 THBS1（血栓素 1）的强烈抑制。尽管肽加工相关基因级联上调，但 PMA/离子霉素刺激后观察到 MHC Ⅰ类基因的抑制。两种刺激后均观察到 MHC Ⅱ类基因的抑制。我们的结果提供了初步数据，表明映射到 SLA 复合体的反义转录本在免疫反应中可能具有作用。
SLA-RI/NRSP8-13K 芯片被发现能够准确破译 PBMCs 的两种不同免疫反应激活，表明它是进一步研究猪免疫和疾病抵抗力的有价值工具。转录组分析显示，根据刺激剂的不同，免疫反应具有特定和共同的特征，这增加了对猪免疫的了解。
