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Remote ischemic conditioning causes CD4 T cells shift towards reduced cell-mediated inflammation

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Abstract

Purpose

Necrotizing enterocolitis (NEC) is a gastrointestinal disease in neonates that is associated with immune-mediated intestinal inflammation. Remote ischemic conditioning (RIC) applied to a limb has been shown to be protective against experimental NEC. In this study, we explore the immune cell-mediated response involved in NEC and the immunomodulatory effects of RIC in an experimental mouse model of the disease.

Methods

NEC was induced in C57BL/6 mice (ethical approval #58119) pups on postnatal day5 (p5) using gavage hyperosmolar formula, lipopolysaccharide, and hypoxia. RIC consisted of 4 cycles of 5 min ischemia followed by 5 min reperfusion of the right hindlimb during NEC induction on p6 and p8. Breastfed mice were used as control. The mice were sacrificed on p9, with ileal tissue evaluated for inflammatory cytokines and by characterization of T-cell populations.

Results

NEC mice had increased number of CD4+ cells indicating an accumulation of T-cells in the mesenchyme of the NEC ileum. Compared to control, NEC pups had upregulated expression pro-inflammatory cytokines (GATA3, IFNγ, IL1β, IL6, IL17, IL22, and TNFα) and reduced anti-inflammatory cytokine (TGFβ). In NEC, there was also a shift in the balance of Treg/Th17 cells towards Th17. Compared to NEC alone, RIC during the course of NEC resulted in reduction of pro-inflammatory cytokines (GATA3, IFNγ, IL1β, IL6, IL17, IL22, and TNFα), increase in anti-inflammatory cytokine TGFβ and concomitant shift back of Th17 cells towards Treg cells.

Conclusion

In experimental NEC, remote ischemic conditioning reduces the production of pro-inflammatory markers and increases the production of anti-inflammatory markers. In addition, during NEC, RIC reverses the imbalance of Treg/Th17 providing support for its effect on cell-mediated inflammation. RIC is a non-invasive physical maneuver that can have a significant beneficial effect in reducing the inflammation seen in NEC.

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Acknowledgements

AP is the recipient of a Canadian Institutes of Health Research (CIHR) Foundation Grant 353857. The funding had no impact on study design, data collection, analysis, interpretation, the writing of the report, or the decision to submit the paper for publication.

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Author notes

  1. Mashriq Alganabi and George Biouss contributed equally.

Authors and Affiliations

  1. Division of General and Thoracic Surgery, Translational Medicine Program, The Hospital for Sick Children, University of Toronto, 555 University Ave, Toronto, ON, M5G 1X8, Canada

    Mashriq Alganabi, George Biouss, Niloofar Ganji, Masaya Yamoto, Carol Lee, Bo Li & Agostino Pierro

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  1. Mashriq Alganabi
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  2. George Biouss
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Correspondence to Agostino Pierro.

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Alganabi, M., Biouss, G., Ganji, N. et al. Remote ischemic conditioning causes CD4 T cells shift towards reduced cell-mediated inflammation. Pediatr Surg Int 38, 657–664 (2022). https://doi.org/10.1007/s00383-022-05093-3

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