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Limosilactobacillus reuteri metabolites modulate immune pathways and intestinal barrier repair after 5 fluorouracil exposure
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  • Published: 02 April 2026

Limosilactobacillus reuteri metabolites modulate immune pathways and intestinal barrier repair after 5 fluorouracil exposure

  • Gintare Lasaviciute1,
  • Marta López Plana1 na1,
  • Sofia Sundberg Örtegren1 na1,
  • Sevasteia Telli1 na2,
  • Symeon Kourmoulakis1 na2,
  • Ludwig Ermann Lundberg2,3,
  • Kenny Lidberg1,
  • Oshadi Peiris1,
  • Indranil Sinha1,
  • Ann-Beth Jonsson1,
  • Stefan Roos2,3,
  • Anna Nilsson4,
  • Manuel Mata Forsberg1 na3 &
  • …
  • Eva Sverremark-Ekström1 na3 

Scientific Reports , Article number:  (2026) Cite this article

We are providing an unedited version of this manuscript to give early access to its findings. Before final publication, the manuscript will undergo further editing. Please note there may be errors present which affect the content, and all legal disclaimers apply.

Subjects

  • Cancer
  • Cell biology
  • Immunology
  • Microbiology

Abstract

Antimetabolites such as 5 fluorouracil are known to induce inflammation in the gut and oral cavity, underscoring the need for strategies that mitigate chemotherapy-associated toxicity. The aim of this study was to determine whether secreted components from the probiotic bacterium Limosilactobacillus reuteri DSM 17938, specifically cell-free supernatant, exopolysaccharides, and extracellular membrane vesicles, can support epithelial barrier recovery following 5 fluorouracil-induced injury. Exposure to 5 fluorouracil impaired viability, metabolic activity, and barrier integrity, and shifted the functional responses of Caco-2 cells toward increased inflammation. Stimulation with exopolysaccharides after removal of 5 fluorouracil significantly improved barrier integrity in both enterocyte-like Caco-2 cells and primary human intestinal epithelial cells, while paradoxically inducing an inflammatory protein profile in the enterocyte-like cells. Transcriptomic analysis revealed that exopolysaccharides modulate gene programs associated with extracellular matrix organization and structural remodelling. Furthermore, cell-free supernatant, membrane vesicles, and exopolysaccharides differentially influenced monocyte polarization pathways when monocytes were cultured with supernatant from 5 fluorouracil-exposed Caco-2 cells. Together, these findings demonstrate that bacterial metabolites such as exopolysaccharides influence intestinal barrier recovery upon inflammation and activate immune cell recruitment that could have consequences for the intestinal epithelial integrity during inflammation.

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Data availability

The datasets generated and/or analysed during the current study will be deposited in the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) upon acceptance of the manuscript and will be made publicly available immediately after publication. Other data is provided within the manuscript or supplemental information files.

Abbreviations

5 FU:

5 Fluorouracil

BEA:

Bioinformatics and expression analysis

CFS:

Cell-free supernatant

CLDN1:

Claudin-1

DEGs:

Differentially expressed genes

DMEM:

Dulbecco’s modified eagle medium

Doxo:

Doxorubicin

EMEM:

Essential medium eagle

EPS:

Exopolysaccharides

ETEC:

Enterotoxigenic Escherichia coli

FITC:

Fluorescein isothiocyanate

GO:

Gene ontology

hSIEC:

human small intestinal epithelial cells

IBD:

Inflammatory bowel disease

IEC:

Intestinal epithelial cells

LGG:

Lacticaseibacillus rhamnosus GG

LR:

Limosilactobacillus reuteri DSM 17938

LTA:

Lipoteichoic acid

MFI:

Mean fluorescent intensity

MRS:

De Man, Rogosa and Sharpe

MV:

Membrane vesicles

NK:

natural killer

OCLN:

Occludin

PBMC:

Peripheral blood mononuclear cells

Papp:

Apparent permeability coefficient

PC:

Principal components

PCA:

Principal component analysis

PRR:

Pattern recognition receptor

RA:

Retinoic acid

TJP:

Tight junction proteins

TEER:

Transepithelial electrical resistance

ZO-1:

Zonula occludens-1

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Acknowledgements

We thank the Imaging Facility at Stockholm University (IFSU) for assistance with confocal microscopy and the BEA core facility at NEO, Karolinska Institutet (Huddinge). We also thank Punya Pallabi Mishra (Department of Molecular Sciences, Swedish University of Agriculture Sciences) for her help with EPS quantification analysis. Finally, we thank Ymke de Jong for her help in analyzing RNA sequencing data.

Funding

Open access funding provided by Stockholm University. This study was supported by the Swedish Research Council under Grant (Dnr 2020 − 01839 and 2023–02616 to ESE); the Swedish Cancer Society under Grant (Dnr CAN 2017/460, 2020 − 1117 and 23 2985Pj); the Cancer and Allergy Foundation; the Mjölkdroppen Foundation; BioGaia; and Stockholm University.

Author information

Author notes

  1. Marta López Plana and Sofia Sundberg Örtegren contributed equally to this work.

  2. Sevasteia Telli and Symeon Kourmoulakis contributed equally to this work.

  3. Manuel Mata Forsberg and Eva Sverremark-Ekström: Shared senior authorship.

Authors and Affiliations

  1. Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Stockholm, 106 91, Sweden

    Gintare Lasaviciute, Marta López Plana, Sofia Sundberg Örtegren, Sevasteia Telli, Symeon Kourmoulakis, Kenny Lidberg, Oshadi Peiris, Indranil Sinha, Ann-Beth Jonsson, Manuel Mata Forsberg & Eva Sverremark-Ekström

  2. Department of Molecular Sciences, Uppsala BioCenter, Swedish University of Agricultural Sciences, Uppsala, Sweden

    Ludwig Ermann Lundberg & Stefan Roos

  3. BioGaia, Stockholm, Sweden

    Ludwig Ermann Lundberg & Stefan Roos

  4. Department of Women’s and Children’s Health, Pediatric Oncology Unit, Karolinska Institute, Stockholm, Sweden

    Anna Nilsson

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Contributions

Gintare Lasaviciute: Conceptualization, methodology, Project administration, investigation, data curation, formal analysis, visualization, writing-original draft, writing-review & editing. Marta López Plana: Investigation. Sofia Sundberg Örtegren: Investigation. Sevasteia Telli: Investigation. Symeon Kourmoulakis: Investigation, writing-review & editing. Ludwig Ermann Lundberg: Resources, writing-review & editing. Kenny Lidberg: Investigation. Oshadi Peiris: Investigation. Indranil Sinha: Data curation, software, formal analysis. Ann-Beth Jonsson: Resources, supervision. Stefan Roos: Resources, writing-review & editing. Anna Nilsson: Conceptualization, funding, writing-review & editing. Manuel Mata Forsberg: Investigation, methodology, validation, supervision, writing-review & editing. Eva Sverremark-Ekström: Conceptualization, project administration, esources, supervision, validation, funding, writing-review & editing.

Corresponding author

Correspondence to Eva Sverremark-Ekström.

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Competing interests

Ludwig Ermann Lundberg and Stefan Roos are employees of BioGaia AB. Eva Sverremark-Ekström has received honoraria for lectures and a research grant for BioGaia AB.

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Lasaviciute, G., López Plana, M., Sundberg Örtegren, S. et al. Limosilactobacillus reuteri metabolites modulate immune pathways and intestinal barrier repair after 5 fluorouracil exposure. Sci Rep (2026). https://doi.org/10.1038/s41598-026-45524-y

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  • Received: 23 December 2025

  • Accepted: 19 March 2026

  • Published: 02 April 2026

  • DOI: https://doi.org/10.1038/s41598-026-45524-y

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Keywords

  • Chemotherapy
  • Probiotics
  • Limosilactobacillus reuteri
  • Exopolysaccharides
  • Extracellular membrane vesicles
  • Epithelial cells integrity
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