Immune Modulation in Peripheral Blood of Cystic Fibrosis Patients Following Ex Vivo Co-Culture with Mesenchymal Stem...

Cystic fibrosis (CF) is an inherited multisystemic disease. Despite advances in treatment, many patients still experience progressive lung dysfunction. Dental follicle-derived mesenchymal stem cells (DF-MSCs) possess significant immunomodulatory potential in inflammatory airway diseases. We evaluated the effects of DF-MSCs on lymphocyte proliferation, CD4⁺CD25⁺FoxP3⁺ regulatory T cell (Treg) frequency, and cytokine responses in peripheral blood mononuclear cells (PBMCs) from CF patients.

PBMCs from 20 CF patients and 20 matched healthy controls were isolated by density gradient centrifugation, stimulated with a CD3/CD28 T-cell-activating antibody cocktail (CD-mix), and co-cultured ex vivo with cryopreserved DF-MSCs. Lymphocyte proliferation was assessed by carboxyfluorescein succinimidyl ester (CFSE)-based flow cytometry, and Tregs were analyzed by flow cytometry. Cytokine levels in culture supernatants were quantified using a multiplex immunoassay.

DF-MSC co-culture significantly suppressed lymphocyte proliferation in CF PBMCs. Treg frequency significantly increased in the stimulated CF samples following MSC co-culture. Following co-culture, levels of tumor necrosis factor-alpha (TNF-α), interleukin-8 (IL-8), and IL-23 decreased. In CF samples, levels of interferon-alpha 2 (IFN-α2), monocyte chemotactic protein-1 (MCP-1), interleukin-12 (IL-12), interleukin-18 (IL-18), and interleukin-33 (IL-33) were elevated.

DF-MSCs reduced lymphocyte proliferation, increased Treg frequency, and regulated cytokine levels in CF, supporting their potential to restore immune balance.