Extended Data Fig. 6: Patients with coeliac disease develop IgA autoantibodies to ameloblast-specific proteins and intestinal antigens.
From: Autoimmune amelogenesis imperfecta in patients with APS-1 and coeliac disease
a, ELISA analyses measuring the reactivity of IgA specific to the indicated enamel proteins in sera from paediatric patients with coeliac disease (n = 24, n = 23 for AMBN) or control individuals without coeliac disease (n = 21). b, ELISA analyses measuring the reactivity of IgA/IgG1 specific to the indicated enamel proteins in sera from adult patients with coeliac disease (n = 48, n = 39 for AMBN and AMTN) or healthy controls (n = 14, n = 48 for FAM20A, n = 6 for AMBN and AMTN). c, Correlation between IgA reactivity to TGM2 vs IgA reactivity to the indicated enamel proteins in paediatric patients with coeliac disease (n = 24) (black dots) and control individuals without coeliac disease (n = 21) (blue dots). d, Representative immunofluorescence microscopy images of Rag1−/− mouse jaw sections stained with sera (green) from patients with coeliac disease (upper panel) or healthy control (lower panel), amelogenin-specific antibodies (red) and DAPI (blue). Scale bar 50 µm. e, ELISA analysis measuring the titre levels of TGM2-specific-sera (IgA) isolated from paediatric patients with coeliac disease (n = 24) or control individuals without coeliac disease (n = 21). f, Representative immunofluorescence microscopy images of Rag1−/− mouse small intestine sections stained with sera (green) from patients with coeliac disease (upper panel) or healthy control (lower panel), TGM2-specific antibodies (red) and DAPI (blue). Scale bar 50 µm. g, Diaminobenzidine (DAB) staining with anti-TGM2 specific antibody showing localization of TGM2 in the developing tooth (IEE=inner enamel epithelium; PreSecA= presecretory ameloblast; SecA=secretory ameloblast) of WT animal. Scale bar 50 µm (upper panel), 25 µM (middle panel) and 20 µM (lower panel). h, Representative immunofluorescence microscopy images of Rag1−/− mouse jaw sections stained with sera (green) from patients with coeliac disease (upper panel) or healthy control (lower panel), TGM2-specific antibodies (red) and DAPI (blue). Scale bar 50 µm. i, Presentation of enamel area (mm2; upper panel) or mineralization (a.u.; lower panel) along the relative (%) incisor location (from lateral to apical part) in representative Tgm2−/− (n = 4) (orange) and WT (n = 4) (blue) mice. Vertical dotted lines indicate a region corresponding to the apical part (that is, the 75th−95th percentile of the incisor’s length). j, Relative expression of LAMB3 gene in different human tissues, based on scRNA-seq datasets publicly available at the human protein atlas database. k, Representative immunofluorescence microscopy images of Rag1−/− mouse small intestine sections stained with sera (green) isolated from patients with coeliac disease celiac (upper panel) or healthy control (lower panel), laminin beta subunit 3-specific antibodies (red), TGM2-specific antibodies (yellow) and DAPI (blue). Scale bar 50 µm. n refers to the number of biologically independent mice/patients per genotype or group. Data are representative of at least two independent experiments. Data shown as mean ± s.e.m. Data were analysed using two-sided Fisher exact test (a, b and e), two-sided Pearson R correlation test (c).
