CFTR regulates B cell activation and lymphoid follicle development

Francesca Polverino,Caroline A Owen,Craig Gerard,Manuela Cernadas,Norma P Gerard,Joselyn Rojas Quintero,Bao Lu,Avignat S Patel,Sara O Vargas

doi:10.1186/s12931-019-1103-1

Abstract

BackgroundCystic fibrosis (CF) is an inherited disorder caused by mutations in the CF transmembrane conductance regulator (CFTR) gene that promotes persistent lung infection and inflammation and progressive loss of lung function. Patients with CF have increased lung lymphoid follicles (LFs) and B cell-activating factor of tumor necrosis factor family (BAFF) that regulates B cell survival and maturation. A direct role for CFTR in B cell activation and disease pathogenesis in CF remains unclear.MethodsThe number of LFs, BAFF+, TLR4+ and proliferation marker Ki67+ B cells in lung explants or resections from subjects with CF and normal controls was quantified by immunostaining. The role of CFTR in B cell activation and LF development was then examined in two independent cohorts of uninfected CFTR-deficient mice (Cftr−/−) and wild type controls. The number of lung LFs, B cells and BAFF+, CXCR4+, immunoglobulin G+ B cells was examined by immunostaining. Lung and splenocyte B cell activation marker and major histocompatibility complex class II (MHC class II) expression was quantified by flow cytometry. Inflammatory cytokine levels were measured in supernatants from isolated B cells from Cftr−/− and wild type mice stimulated in vitro with Pseudomonas aeruginosa lipopolysaccharide (LPS).ResultsThere was a significant increase in well-formed LFs in subjects with CF compared to normal controls. Increased B cell activation and proliferation was observed in lung LFs from CF subjects as was quantified by a significant increase in B cell BAFF, TLR4 and Ki67 expression. Uninfected Cftr−/− mice had increased lung LFs and BAFF+ and CXCR4+ B cells compared to wild type controls. Lung B cells isolated from uninfected Cftr−/− mice demonstrated increased MHC class II expression. In vitro, isolated B cells from Cftr−/− mice produced increased IL-6 when stimulated with LPS compared to wild type controls.ConclusionsThese data support a direct role for CFTR in B cell activation, proliferation and inflammatory cytokine production that promotes lung LF follicle development in cystic fibrosis.

Highlights

Cystic fibrosis (CF) is an inherited disorder caused by mutations in the CF transmembrane conductance regulator (CFTR) gene that promotes persistent lung infection and inflammation and progressive loss of lung function
CF arises from mutations in the CF transmembrane conductance regulator (CFTR) that lead to impaired chloride and bicarbonate transport
Patient characteristics Lung tissue sections were obtained from patients with CF (n = 27) and normal controls (n = 10)

Summary

Introduction

Cystic fibrosis (CF) is an inherited disorder caused by mutations in the CF transmembrane conductance regulator (CFTR) gene that promotes persistent lung infection and inflammation and progressive loss of lung function. Patients with CF have increased lung lymphoid follicles (LFs) and B cell-activating factor of tumor necrosis factor family (BAFF) that regulates B cell survival and maturation. Methods: The number of LFs, BAFF+, TLR4+ and proliferation marker Ki67+ B cells in lung explants or resections from subjects with CF and normal controls was quantified by immunostaining. Impaired CFTR epithelial cell function in patients with CF leads to viscous mucus, impaired mucociliary clearance and airway colonization with pathogenic bacteria, especially Pseudomonas aeruginosa (P. aeruginosa). These changes in airway biology lead to progressive lung damage and respiratory insufficiency. CFTR expression on both innate and adaptive immune cells has been increasingly appreciated

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