We previously identified a Neisseria flavescens strain in the duodenum of celiac disease (CD) patients that induced immune inflammation in ex vivo duodenal mucosal explants and in CaCo-2 cells. We also found that vesicular trafficking was delayed after the CD-immunogenic P31-43 gliadin peptide-entered CaCo-2 cells and that Lactobacillus paracasei CBA L74 (L. paracasei-CBA) supernatant reduced peptide entry. In this study, we evaluated if metabolism and trafficking was altered in CD-N. flavescens-infected CaCo-2 cells and if any alteration could be mitigated by pretreating cells with L. paracasei-CBA supernatant, despite the presence of P31-43. We measured CaCo-2 bioenergetics by an extracellular flux analyser, N. flavescens and P31-43 intracellular trafficking by immunofluorescence, cellular stress by TBARS assay, and ATP by bioluminescence. We found that CD-N. flavescens colocalised more than control N. flavescens with early endocytic vesicles and more escaped autophagy thereby surviving longer in infected cells. P31-43 increased colocalisation of N. flavescens with early vesicles. Mitochondrial respiration was lower (P < .05) in CD-N. flavescens-infected cells versus not-treated CaCo-2 cells, whereas pretreatment with L. paracasei-CBA reduced CD-N. flavescens viability and improved cell bioenergetics and trafficking. In conclusion, CD-N. flavescens induces metabolic imbalance in CaCo-2 cells, and the L. paracasei-CBA probiotic could be used to correct CD-associated dysbiosis.

中文翻译：

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乳糜泻相关的 Neisseria flavescens 降低 CaCo-2 上皮细胞的线粒体呼吸：副干酪乳杆菌 CBA L74 对细菌诱导的细胞失衡的影响。

我们之前在乳糜泻 (CD) 患者的十二指肠中发现了一种在离体十二指肠粘膜外植体和 CaCo-2 细胞中诱导免疫炎症的 Neisseria flavescens 菌株。我们还发现，在 CD 免疫原性 P31-43 麦胶蛋白肽进入 CaCo-2 细胞后，囊泡运输被延迟，并且副干酪乳杆菌 CBA L74 (L. paracasei-CBA) 上清液减少了肽进入。在这项研究中，我们评估了 CD-N 中的代谢和运输是否发生了改变。尽管存在 P31-43，但通过用副干酪乳杆菌-CBA 上清液预处理细胞可以减轻苦参感染的 CaCo-2 细胞以及如果有任何改变。我们通过细胞外通量分析仪测量 CaCo-2 生物能量学，通过免疫荧光测量苦参和 P31-43 细胞内运输，通过 TBARS 测定测量细胞应激，通过生物发光测量 ATP。我们发现 CD-N。与对照 N. flavescens 相比，flavescens 与早期内吞囊泡和更多逃脱的自噬共定位，从而在受感染的细胞中存活更长时间。P31-43 增加了 N. flavescens 与早期囊泡的共定位。CD-N 中的线粒体呼吸较低 (P < .05)。苦参感染的细胞与未处理的 CaCo-2 细胞相比，而用副干酪乳杆菌-CBA 预处理减少了 CD-N。苦参的活力和改善的细胞生物能量学和运输。总之，CD-N。苦参诱导 CaCo-2 细胞的代谢失衡，L. paracasei-CBA 益生菌可用于纠正 CD 相关的生态失调。具有早期囊泡的苦参。CD-N 中的线粒体呼吸较低 (P < .05)。苦参感染的细胞与未处理的 CaCo-2 细胞相比，而用副干酪乳杆菌-CBA 预处理减少了 CD-N。苦参的活力和改善的细胞生物能量学和运输。总之，CD-N。苦参诱导 CaCo-2 细胞的代谢失衡，L. paracasei-CBA 益生菌可用于纠正 CD 相关的生态失调。具有早期囊泡的苦参。CD-N 中的线粒体呼吸较低 (P < .05)。苦参感染的细胞与未处理的 CaCo-2 细胞相比，而用副干酪乳杆菌-CBA 预处理减少了 CD-N。苦参的活力和改善的细胞生物能量学和运输。总之，CD-N。苦参诱导 CaCo-2 细胞的代谢失衡，L. paracasei-CBA 益生菌可用于纠正 CD 相关的生态失调。