Celiac disease-associated Neisseria flavescens decreases mitochondrial respiration in CaCo-2 epithelial cells: Impact of Lactobacillus paracasei CBA L74 on bacterial-induced cellular imbalance

Cell Microbiol. 2019 Aug;21(8):e13035. doi: 10.1111/cmi.13035. Epub 2019 May 20.

Abstract

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.

Keywords: CaCo-2 cells; L. paracasei CBA L74 probiotic; Neisseria flavescens; P31-43 gliadin peptide; celiac disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / agonists
  • Adenosine Triphosphate / metabolism
  • Autophagosomes / drug effects
  • Autophagosomes / metabolism
  • Autophagosomes / microbiology
  • Autophagy / drug effects
  • Autophagy / genetics
  • Caco-2 Cells
  • Celiac Disease / metabolism
  • Celiac Disease / microbiology
  • Celiac Disease / therapy
  • Culture Media, Conditioned / pharmacology
  • Dysbiosis / metabolism
  • Dysbiosis / microbiology
  • Dysbiosis / therapy
  • Gene Expression
  • Gliadin / antagonists & inhibitors
  • Gliadin / pharmacology
  • Host-Pathogen Interactions / drug effects
  • Host-Pathogen Interactions / genetics
  • Humans
  • Lacticaseibacillus paracasei / chemistry*
  • Lacticaseibacillus paracasei / physiology
  • Lysosomal-Associated Membrane Protein 2 / genetics
  • Lysosomal-Associated Membrane Protein 2 / metabolism
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism
  • Mitochondria / drug effects*
  • Mitochondria / metabolism
  • Neisseria / drug effects*
  • Neisseria / genetics
  • Neisseria / growth & development
  • Neisseria / pathogenicity
  • Oxidative Phosphorylation / drug effects*
  • Peptide Fragments / antagonists & inhibitors
  • Peptide Fragments / pharmacology
  • Probiotics / pharmacology*
  • Thiobarbituric Acid Reactive Substances / metabolism
  • Transport Vesicles / drug effects
  • Transport Vesicles / metabolism
  • Transport Vesicles / ultrastructure
  • Vesicular Transport Proteins / genetics
  • Vesicular Transport Proteins / metabolism

Substances

  • Culture Media, Conditioned
  • LAMP2 protein, human
  • Lysosomal-Associated Membrane Protein 2
  • MAP1LC3A protein, human
  • Microtubule-Associated Proteins
  • Peptide Fragments
  • Thiobarbituric Acid Reactive Substances
  • Vesicular Transport Proteins
  • early endosome antigen 1
  • gliadin peptide (31-43)
  • Adenosine Triphosphate
  • Gliadin

Supplementary concepts

  • Neisseria flavescens