Fig. 2: lnc13 maintains oral tolerance to gluten in HLA-DQ8–restricted mice.
From: The long noncoding RNA lnc13 restrains inflammatory responses to maintain oral tolerance to gluten
a, Schematic of the gluten challenge protocol in HLA-DQ8 transgenic mice. Mice were weaned onto a gluten-free diet, then switched to standard chow and gavaged with 20 mg gliadin every other day for 45 days. b,c, Relative RNA expression of type 1 cytokines Ifng (b; P = 0.00351387) and Il12b (c; P = 0.00207621) in terminal ileum tissue from gluten-free or gluten-challenged mice, measured by RT–qPCR. Relative RNA levels are calculated in reference to Rpl32 housekeeping gene. n = 3–6 mice, across four independent experiments. d, lnc13 expression in terminal ileum from gluten-free versus gluten-challenged DQ8 mice, showing decreased expression upon gluten exposure. Relative RNA levels are calculated in reference to Rpl32 housekeeping gene. n = 3–6, across four independent experiments. P = 0.00023608. e–g, Representative flow cytometry plots (left) and quantification (right) showing frequencies of IFN-γ+ cells from live CD45+TCRb+B220− cells, where quantifications show those further gated on CD8+CD4− (P = 0.00013518) or CD4+CD8− (P = 0.00047811) (e), frequencies of IL-12+CD11b+ cells from live CD45+TCRb−B220− cells (P = 0.00031403) (f) and frequencies of FOXP3+ cells (P = 0.03630467) from live CD45+TCRb+B220−CD4+CD8− cells (g) in the small intestinal lamina propria (Lp) of gluten-challenged lnc13−/−-DQ8 and WT-DQ8 mice. n = 6 mice, across four independent experiments. h, Pathway enrichment analysis of DEGs in whole ileum (‘tissue’) and Lp lymphocytes (LPLs) from lnc13−/−-DQ8 versus WT-DQ8 mice after gluten challenge, and duodenal biopsies from patients with CeD versus healthy controls (GSE134900). Shared pathways include immune response and cytokine-mediated signaling. i, Expression of selected proinflammatory and B cell-associated genes in the ileum of lnc13−/−-DQ8 versus WT-DQ8 mice by RT–qPCR. Relative RNA levels (ΔΔCt) are calculated in reference to Rpl32 and then normalized to the average of WT-DQ8 samples. n = 6 across three independent experiments. P values for Il1a, Il21, Csf2, Il2, Tnfa, Il27, Il13, Il1b, Il17a, Hprt, Il7, Cd138 and Blimp1 are 0.006489, 0.004429, 0.001036, 0.000873, 0.030904, 0.023022, 0.0517, 0.0078, 0.035474, 0.299992923, 0.61115255, 0.20111989 and 0.02767208, respectively. An unpaired, two-tailed, Student’s t-test was used for comparison (b–g,i). *P < 0.05,**P < 0.01, ***P < 0.001, NS, not significant. In scatter-plots (b,c,e–g,i), samples from lnc13−/−-DQ8 (KO-DQ8) mice are represented by red dots and those from lnc13+/+-DQ8 (WT-DQ8) mice are represented by blue dots. Lines represent mean ± s.e.m. Image of mouse in a generated by BioRender; Yang-Fischer, R. https://biorender.com/hz505m6 (2026).
