Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Avenida do Café, s/nº, Monte Alegre, Ribeirão Preto, SP, CEP 14040-903, Brazil.
World J Microbiol Biotechnol. 2021 Jan 18;37(2):31. doi: 10.1007/s11274-021-02996-6.
Spheroids of intestinal cells (Caco-2) were used to evaluate the adhesion/invasion ability of Listeria monocytogenes (pathogen) and Lactobacillus sakei 1 (potential probiotic). Besides, transcriptomic analyses of Caco-2 cells in three dimensional cultures were done, with the aim of revealing possible host-foodborne bacteria interactions. Result of adhesion assay for L. monocytogenes in Caco-2 spheroids was 22.86 ± 0.33%, but it was stimulated in acidic pH (4.5) and by the presence of 2% sucrose (respectively, 32.56 ± 1.35% and 33.25 ± 1.26%). Conversely, the invasion rate of L. monocytogenes was lower at pH 4.5, in comparison with non-stressed controls (18.89 ± 1.05% and 58.65 ± 0.30%, respectively). L. sakei 1 adhered to Caco-2 tridimensional cell culture (27.30 ± 2.64%), with no invasiveness. There were 19 and 21 genes down and upregulated, respectively, in tridimensional Caco-2 cells, upon infection with L. monocytogenes, which involved immunity, apoptosis; cytoprotective responses, cell signalling-regulatory pathways. It was evidenced despite activation or deactivation of several pathways in intestinal cells to counteract infection, the pathogen was able to hijack many host defense mechanisms. On the other hand, the probiotic candidate L. sakei 1 was correlated with decreased transcription of two genes in Caco-2 cells, though it stimulated the expression of 14 others, with diverse roles in immunity, apoptosis, cytoprotective response and cell signalling-regulatory pathways. Our data suggest the use of tridimensional cell culture to mimic the intestinal epithelium is a good model for gathering broad information on the putative mechanisms of interaction between host and bacteria of importance for food safety, which can serve as a basis for further in-depth investigation.
肠细胞球体（Caco-2）用于评估李斯特菌（病原体）和清酒乳杆菌 1（潜在益生菌）的黏附/侵袭能力。此外，还对三维培养的 Caco-2 细胞进行了转录组分析，目的是揭示可能的宿主-食源性病原体相互作用。李斯特菌在 Caco-2 球体中的黏附率为 22.86±0.33%，但在酸性 pH（4.5）和 2%蔗糖存在的情况下会受到刺激（分别为 32.56±1.35%和 33.25±1.26%）。相反，李斯特菌的侵袭率在 pH 4.5 时低于非应激对照（分别为 18.89±1.05%和 58.65±0.30%）。清酒乳杆菌 1 黏附于 Caco-2 三维细胞培养物（27.30±2.64%），无侵袭性。感染李斯特菌后，三维 Caco-2 细胞中分别有 19 个和 21 个基因下调和上调，涉及免疫、凋亡；细胞保护反应、细胞信号转导调节途径。有证据表明，尽管肠道细胞中激活或失活了几种途径来对抗感染，但病原体仍能够劫持许多宿主防御机制。另一方面，益生菌候选物清酒乳杆菌 1 与 Caco-2 细胞中两个基因的转录减少有关，尽管它刺激了另外 14 个基因的表达，这些基因在免疫、凋亡、细胞保护反应和细胞信号转导调节途径中具有不同的作用。我们的数据表明，使用三维细胞培养来模拟肠道上皮是收集有关宿主与食源性病原体之间相互作用的潜在机制的广泛信息的良好模型，这可以作为进一步深入研究的基础。
