Journal Article

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

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

2024

Overview

In 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. We 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. The 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. We 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. B-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.

• Publisher: Frontiers Media SA,Frontiers,Frontiers Media S.A
• Subject: 3D coculture / Adult / B-cell / B-Lymphocytes - immunology / B-Lymphocytes - metabolism / Biopsy / Cell activation / Cell Communication - immunology / Cells, Cultured / Cloning / Coculture Techniques / Cytokines / Cytokines - metabolism / Extracellular matrix / Female / fibroblast / Fibroblasts / Fibroblasts - immunology / Fibroblasts - metabolism / fibrosis / Humans / Immunology / Infiltration / Inflammation / Keratinocytes / Keratinocytes - immunology / Keratinocytes - metabolism / Life Sciences / Ligands / Lymphocyte Activation - immunology / Lymphocytes B / Male / Memory cells / Middle Aged / Monoclonal antibodies / Pathophysiology / Patients / RNA polymerase / Scleroderma / Scleroderma, Systemic - immunology / Scleroderma, Systemic - metabolism / Scleroderma, Systemic - pathology / Skin / Skin - immunology / Skin - metabolism / Skin - pathology / Systemic sclerosis / Transcriptome / Transcriptomes / Transcriptomics