Frontiers in Immunology (Aug 2024)

Characteristics and impact of infiltration of B-cells from systemic sclerosis patients in a 3D healthy skin model

  • Mathilde Le Maître,
  • Thomas Guerrier,
  • Aurore Collet,
  • Aurore Collet,
  • Mehdi Derhourhi,
  • Mehdi Derhourhi,
  • Jean-Pascal Meneboo,
  • Bénédicte Toussaint,
  • Bénédicte Toussaint,
  • Amélie Bonnefond,
  • Amélie Bonnefond,
  • Amélie Bonnefond,
  • Céline Villenet,
  • Shéhérazade Sebda,
  • Antonino Bongiovanni,
  • Meryem Tardivel,
  • Myriam Simon,
  • Manel Jendoubi,
  • Blanche Daunou,
  • Alexis Largy,
  • Martin Figeac,
  • Sylvain Dubucquoi,
  • Sylvain Dubucquoi,
  • David Launay,
  • David Launay

DOI
https://doi.org/10.3389/fimmu.2024.1373464
Journal volume & issue
Vol. 15

Abstract

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IntroductionIn systemic sclerosis (SSc), B-cells are activated and present in the skin and lung of patients where they can interact with fibroblasts. The precise impact and mechanisms of the interaction of B-cells and fibroblasts at the tissular level are poorly studied.ObjectiveWe investigated the impact and mechanisms of B-cell/fibroblast interactions in cocultures between B-cells from patients with SSc and 3-dimensional reconstituted healthy skin model including fibroblasts, keratinocytes and extracellular matrix.MethodsThe quantification and description of the B-cell infiltration in 3D cocultures were performed using cells imagery strategy and cytometry. The effect of coculture on the transcriptome of B-cells and fibroblasts was studied with bulk and single-cell RNA sequencing approaches. The mechanisms of this interaction were studied by blocking key cytokines like IL-6 and TNF.ResultsWe showed a significant infiltration of B-cells in the 3D healthy skin model. The amount but not the depth of infiltration was higher with B-cells from SSc patients and with activated B-cells. B-cell infiltrates were mainly composed of naïve and memory cells, whose frequencies differed depending on B-cells origin and activation state: infiltrated B-cells from patients with SSc showed an activated profile and an overexpression of immunoglobulin genes compared to circulating B-cells before infiltration. Our study has shown for the first time that activated B-cells modified the transcriptomic profile of both healthy and SSc fibroblasts, toward a pro-inflammatory (TNF and IL-17 signaling) and interferon profile, with a key role of the TNF pathway.ConclusionB-cells and 3D skin cocultures allowed the modelization of B-cells infiltration in tissues observed in SSc, uncovering an influence of the underlying disease and the activation state of B-cells. We showed a pro-inflammatory effect on skin fibroblasts and pro-activation effect on infiltrating B-cells during coculture. This reinforces the role of B-cells in SSc and provide potential targets for future therapeutic approach in this disease.

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