The use of in vitro systems that allow efficient selection of probiotic candidates with immunomodulatory properties could significantly minimize the use of experimental animals. In this work, we generated an in vitro immunoassay system based on porcine intestinal epithelial (PIE) cells and dextran sodium sulfate (DSS) administration that could be useful for the selection and characterization of potential probiotic strains to be used in inflammatory bowel disease (IBD) patients. Our strategy was based on two fundamental pillars: on the one hand, the capacity of PIE cells to create a monolayer by attaching to neighboring cells and efficiently mount inflammatory responses and, on the other hand, the use of two probiotic bifidobacteria strains that have been characterized in terms of their immunomodulatory capacities, particularly in mouse IBD models and patients. Our results demonstrated that DSS administration can alter the epithelial barrier created in vitro by PIE cells and induce a potent inflammatory response, characterized by increases in the expression levels of several inflammatory factors including TNF-α, IL-1α, CCL4, CCL8, CCL11, CXCL5, CXCL9, CXCL10, SELL, SELE, EPCAM, VCAM, NCF2, and SAA2. In addition, we demonstrated that Bifidobacterium breve M-16V and B. longum BB536 are able to regulate the C-jun N-terminal kinase (JNK) intracellular signalling pathway, reducing the DSS-induced alterations of the in vitro epithelial barrier and differentially regulating the inflammatory response in a strain-dependent fashion. The good correlation between our in vitro findings in PIE cells and previous studies in animal models and IBD patients shows the potential value of our system to select new probiotic candidates in an efficient way.

使用允许有效选择具有免疫调节特性的益生菌候选物的体外系统的使用可以显着减少实验动物的使用。在这项工作中，我们生成了一个基于猪肠上皮 (PIE) 细胞和葡聚糖硫酸钠 (DSS) 给药的体外免疫分析系统，该系统可用于选择和表征用于炎症性肠病 (IBD) 的潜在益生菌菌株） 耐心。我们的策略基于两个基本支柱：一方面，PIE 细胞通过附着在相邻细胞上并有效引发炎症反应来产生单层的能力，另一方面，使用两种益生菌双歧杆菌菌株以它们的免疫调节能力为特征，特别是在小鼠 IBD 模型和患者中。我们的结果表明，DSS 给药可以改变 PIE 细胞在体外产生的上皮屏障并诱导有效的炎症反应，其特征是几种炎症因子的表达水平增加，包括 TNF-α、IL-1α、CCL4、CCL8、CCL11、 CXCL5、CXCL9、CXCL10、SELL、SELE、EPCAM、VCAM、NCF2 和 SAA2。此外，我们证明了短双歧杆菌M-16V和乙。longum BB536 能够调节 C-jun N 末端激酶 (JNK) 细胞内信号通路，减少 DSS 诱导的体外上皮屏障的改变，并以菌株依赖性方式差异调节炎症反应。我们在 PIE 细胞中的体外发现与之前在动物模型和 IBD 患者中的研究之间的良好相关性显示了我们的系统以有效方式选择新的益生菌候选物的潜在价值。