Figure S1.
Analysis of PC longevity and specificity by flow cytometry. (A) Experimental approach. The red cross indicates a GFD. The blue square indicates location of biopsy sampling (duodenum). (B) Single-cell FACS index sorting based on antigen specificity. TG2- and DGP-specific PCs were identified by fluorescent tetramers (SA) loaded with TG2 or DGP. Representative flow cytometry plots and sorting strategy is shown for one CeD patient. Gates used for sorting are outlined in red. (C) Single-cell FACS index sorting for control subjects. PCs of unknown specificity were sorted with a natural composition of PC longevity populations for three controls (left). Equal numbers of short-, intermediate-, and long-lived PCs were sorted from two controls (right) based on CD45/CD19 surface marker expression. Flow cytometry plots from one control are shown. Gates used for sorting are outlined in red. (D) Age distribution in patient groups. (E) Representative flow cytometry plots showing percent TG2-specific and DGP-specific PCs and the longevity markers CD19/CD45 for each specificity for one UCeD, one TCeD, and one control subject. PCs were gated as large, single CD3−CD11c−CD14−CD38high cells. (F) Percentage of PCs identified as being specific for TG2 (left) or DGP (right) in UCeD (n = 15), TCeD (n = 26), and controls (n = 13) by flow cytometry, colored according to the Marsh score of intestinal inflammation (Oberhuber et al., 1999). P values were calculated with an unpaired Wilcoxon rank-sum test. Adjusted P values are shown. **, P < 0.01; ****, P < 0.0001. (G) Percentage of PCs identified as being specific for TG2 (top) or DGP (bottom) in UCeD (n = 15) and TCeD patients (n = 26) as a function of time on a GFD. (H) Percentage of PCs belonging to the short-, intermediate- (CD19−CD45+), and long-lived compartments for DGP-specific PCs and other PCs as a function of treatment duration (years on GFD). UCeD patients are plotted as 0 yr on GFD. The colored lines represent fitted trend using linear regression; shades indicate the 95% confidence interval of the fitted line. Only patients with at least 30 recorded DGP-specific PCs are included (n = 27).

Analysis of PC longevity and specificity by flow cytometry. (A) Experimental approach. The red cross indicates a GFD. The blue square indicates location of biopsy sampling (duodenum). (B) Single-cell FACS index sorting based on antigen specificity. TG2- and DGP-specific PCs were identified by fluorescent tetramers (SA) loaded with TG2 or DGP. Representative flow cytometry plots and sorting strategy is shown for one CeD patient. Gates used for sorting are outlined in red. (C) Single-cell FACS index sorting for control subjects. PCs of unknown specificity were sorted with a natural composition of PC longevity populations for three controls (left). Equal numbers of short-, intermediate-, and long-lived PCs were sorted from two controls (right) based on CD45/CD19 surface marker expression. Flow cytometry plots from one control are shown. Gates used for sorting are outlined in red. (D) Age distribution in patient groups. (E) Representative flow cytometry plots showing percent TG2-specific and DGP-specific PCs and the longevity markers CD19/CD45 for each specificity for one UCeD, one TCeD, and one control subject. PCs were gated as large, single CD3−CD11c−CD14−CD38high cells. (F) Percentage of PCs identified as being specific for TG2 (left) or DGP (right) in UCeD (n = 15), TCeD (n = 26), and controls (n = 13) by flow cytometry, colored according to the Marsh score of intestinal inflammation (Oberhuber et al., 1999). P values were calculated with an unpaired Wilcoxon rank-sum test. Adjusted P values are shown. **, P < 0.01; ****, P < 0.0001. (G) Percentage of PCs identified as being specific for TG2 (top) or DGP (bottom) in UCeD (n = 15) and TCeD patients (n = 26) as a function of time on a GFD. (H) Percentage of PCs belonging to the short-, intermediate- (CD19−CD45+), and long-lived compartments for DGP-specific PCs and other PCs as a function of treatment duration (years on GFD). UCeD patients are plotted as 0 yr on GFD. The colored lines represent fitted trend using linear regression; shades indicate the 95% confidence interval of the fitted line. Only patients with at least 30 recorded DGP-specific PCs are included (n = 27).

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