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Increased Intraepithelial Vα24 Invariant NKT Cells in the Celiac Duodenum.

Montalvillo E, Bernardo D, Martínez-Abad B, Allegretti Y, Fernández-Salazar L, Calvo C, Chirdo FG, Garrote JA, Arranz E - Nutrients (2015)

Bottom Line: The expression of Vα24-Jα18 correlated with Marsh score for the severity of mucosal lesion and also with increased mRNA IFNγ levels. uCD and GFD-CD patients had decreased mRNA expression of FoxP3 but increased expression of Vα24-Jα18, which revealed a CD-like molecular profile.In conclusion, we have found an increased number of iNKT cells in the duodenum from both uCD and GFD-CD patients, irrespective of the mucosal status.A CD-like molecular profile, defined by an increased mRNA expression of Vα24-Jα18 together with a decreased expression of FoxP3, may represent a pro-inflammatory signature of the CD duodenum.

View Article: PubMed Central - PubMed

Affiliation: Mucosal Immunology Lab, IBGM, University of Valladolid-CSIC, Sanz y Forés 3, 47003 Valladolid, Spain. emontalvillo@gmail.com.

ABSTRACT
Celiac Disease (CD) is an interferon (IFN)γ-mediated duodenal hypersensitivity to wheat gluten occurring in genetically predisposed individuals. Gluten-free diet (GFD) leads to a complete remission of the disease. Vα24-restricted invariant NKT (iNKT) cells are important to maintain immune homeostasis in the gut mucosa because of their unique capacity to rapidly produce large quantities of both T-helper (Th)1 and Th2 cytokines upon stimulation. We studied the presence of these cells in the CD duodenum. Duodenal biopsies were obtained from 45 untreated-CD patients (uCD), 15 Gluten Free Diet-CD patients (GFD-CD), 44 non-inflamed non-CD controls (C-controls) and 15 inflamed non-CD controls (I-controls). Two populations from Spain and Argentina were recruited. Messenger RNA (mRNA) expression of Vα24-Jα18 (invariant TCRα chain of human iNKT cells), IFNγ and intracellular transcription factor Forkhead Box P3 (Foxp3), and flow cytometry intraepithelial lymphocyte (IEL) profile were determined. Both uCD and GFD-CD patients had higher Vα24-Jα18 mRNA levels than non-CD controls (I and C-controls). The expression of Vα24-Jα18 correlated with Marsh score for the severity of mucosal lesion and also with increased mRNA IFNγ levels. uCD and GFD-CD patients had decreased mRNA expression of FoxP3 but increased expression of Vα24-Jα18, which revealed a CD-like molecular profile. Increased numbers of iNKT cells were confirmed by flow cytometry within the intraepithelial lymphocyte compartment of uCD and GFD-CD patients and correlated with Vα24-Jα18 mRNA expression. In conclusion, we have found an increased number of iNKT cells in the duodenum from both uCD and GFD-CD patients, irrespective of the mucosal status. A CD-like molecular profile, defined by an increased mRNA expression of Vα24-Jα18 together with a decreased expression of FoxP3, may represent a pro-inflammatory signature of the CD duodenum.

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Duodenal mRNA levels of Vα24-Jα18 and FoxP3 reveal a celiac disease molecular profile. Expression levels of FoxP3 mRNA in duodenal biopsies indicated as the ratio FoxP3/βactin, in arbitrary units (AU) in untreated Celiac Disease (uCD), Gluten Free Diet-CD patients (GFD-CD), inflamed non-CD controls (I-controls) and non-inflamed non-CD controls (C-controls) in the Spanish population (A) and in uCD and C-controls in the Argentinian population (B). Statistical differences are shown (two tailed Mann-Whitney U test; Krustall-Wallis test). Horizontal bars are median values. Duodenal molecular profile: Correlation between expression levels of FoxP3 and Vα24-Jα18 mRNA, in arbitrary units (AU) (C-controls: Spearman r = 0.3495, p value < 0.05; I-controls: Spearman r = −0.4577, p value = n.s; GFD-CD: Spearman r = −0.4858, p value < 0.05; uCD: Spearman r = −0.056, p value = n.s) (C).
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nutrients-07-05444-f004: Duodenal mRNA levels of Vα24-Jα18 and FoxP3 reveal a celiac disease molecular profile. Expression levels of FoxP3 mRNA in duodenal biopsies indicated as the ratio FoxP3/βactin, in arbitrary units (AU) in untreated Celiac Disease (uCD), Gluten Free Diet-CD patients (GFD-CD), inflamed non-CD controls (I-controls) and non-inflamed non-CD controls (C-controls) in the Spanish population (A) and in uCD and C-controls in the Argentinian population (B). Statistical differences are shown (two tailed Mann-Whitney U test; Krustall-Wallis test). Horizontal bars are median values. Duodenal molecular profile: Correlation between expression levels of FoxP3 and Vα24-Jα18 mRNA, in arbitrary units (AU) (C-controls: Spearman r = 0.3495, p value < 0.05; I-controls: Spearman r = −0.4577, p value = n.s; GFD-CD: Spearman r = −0.4858, p value < 0.05; uCD: Spearman r = −0.056, p value = n.s) (C).

Mentions: We next studied FoxP3 mRNA expression in the duodenum as an indirect way of quantifying Treg cells in tissue. CD samples (both uCD and GFD-CD) had decreased FoxP3 expression compared with non-inflamed non-CD-controls (C-controls) (uCD, p < 0.001; GFD-CD, p < 0.001) (Figure 4A). Similar results were found on a second independent analysis of the Argentinian population (uCD, p < 0.001) (Figure 4B). Because CD samples were characterized by increased duodenal mRNA expression of Vα24-Jα18 (Figure 2A,B) and decreased expression of FoxP3 (Figure 4A,B), we studied whether the joint analysis of these two molecules could help us to identify a CD-like molecular profile.


Increased Intraepithelial Vα24 Invariant NKT Cells in the Celiac Duodenum.

Montalvillo E, Bernardo D, Martínez-Abad B, Allegretti Y, Fernández-Salazar L, Calvo C, Chirdo FG, Garrote JA, Arranz E - Nutrients (2015)

Duodenal mRNA levels of Vα24-Jα18 and FoxP3 reveal a celiac disease molecular profile. Expression levels of FoxP3 mRNA in duodenal biopsies indicated as the ratio FoxP3/βactin, in arbitrary units (AU) in untreated Celiac Disease (uCD), Gluten Free Diet-CD patients (GFD-CD), inflamed non-CD controls (I-controls) and non-inflamed non-CD controls (C-controls) in the Spanish population (A) and in uCD and C-controls in the Argentinian population (B). Statistical differences are shown (two tailed Mann-Whitney U test; Krustall-Wallis test). Horizontal bars are median values. Duodenal molecular profile: Correlation between expression levels of FoxP3 and Vα24-Jα18 mRNA, in arbitrary units (AU) (C-controls: Spearman r = 0.3495, p value < 0.05; I-controls: Spearman r = −0.4577, p value = n.s; GFD-CD: Spearman r = −0.4858, p value < 0.05; uCD: Spearman r = −0.056, p value = n.s) (C).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4663572&req=5

nutrients-07-05444-f004: Duodenal mRNA levels of Vα24-Jα18 and FoxP3 reveal a celiac disease molecular profile. Expression levels of FoxP3 mRNA in duodenal biopsies indicated as the ratio FoxP3/βactin, in arbitrary units (AU) in untreated Celiac Disease (uCD), Gluten Free Diet-CD patients (GFD-CD), inflamed non-CD controls (I-controls) and non-inflamed non-CD controls (C-controls) in the Spanish population (A) and in uCD and C-controls in the Argentinian population (B). Statistical differences are shown (two tailed Mann-Whitney U test; Krustall-Wallis test). Horizontal bars are median values. Duodenal molecular profile: Correlation between expression levels of FoxP3 and Vα24-Jα18 mRNA, in arbitrary units (AU) (C-controls: Spearman r = 0.3495, p value < 0.05; I-controls: Spearman r = −0.4577, p value = n.s; GFD-CD: Spearman r = −0.4858, p value < 0.05; uCD: Spearman r = −0.056, p value = n.s) (C).
Mentions: We next studied FoxP3 mRNA expression in the duodenum as an indirect way of quantifying Treg cells in tissue. CD samples (both uCD and GFD-CD) had decreased FoxP3 expression compared with non-inflamed non-CD-controls (C-controls) (uCD, p < 0.001; GFD-CD, p < 0.001) (Figure 4A). Similar results were found on a second independent analysis of the Argentinian population (uCD, p < 0.001) (Figure 4B). Because CD samples were characterized by increased duodenal mRNA expression of Vα24-Jα18 (Figure 2A,B) and decreased expression of FoxP3 (Figure 4A,B), we studied whether the joint analysis of these two molecules could help us to identify a CD-like molecular profile.

Bottom Line: The expression of Vα24-Jα18 correlated with Marsh score for the severity of mucosal lesion and also with increased mRNA IFNγ levels. uCD and GFD-CD patients had decreased mRNA expression of FoxP3 but increased expression of Vα24-Jα18, which revealed a CD-like molecular profile.In conclusion, we have found an increased number of iNKT cells in the duodenum from both uCD and GFD-CD patients, irrespective of the mucosal status.A CD-like molecular profile, defined by an increased mRNA expression of Vα24-Jα18 together with a decreased expression of FoxP3, may represent a pro-inflammatory signature of the CD duodenum.

View Article: PubMed Central - PubMed

Affiliation: Mucosal Immunology Lab, IBGM, University of Valladolid-CSIC, Sanz y Forés 3, 47003 Valladolid, Spain. emontalvillo@gmail.com.

ABSTRACT
Celiac Disease (CD) is an interferon (IFN)γ-mediated duodenal hypersensitivity to wheat gluten occurring in genetically predisposed individuals. Gluten-free diet (GFD) leads to a complete remission of the disease. Vα24-restricted invariant NKT (iNKT) cells are important to maintain immune homeostasis in the gut mucosa because of their unique capacity to rapidly produce large quantities of both T-helper (Th)1 and Th2 cytokines upon stimulation. We studied the presence of these cells in the CD duodenum. Duodenal biopsies were obtained from 45 untreated-CD patients (uCD), 15 Gluten Free Diet-CD patients (GFD-CD), 44 non-inflamed non-CD controls (C-controls) and 15 inflamed non-CD controls (I-controls). Two populations from Spain and Argentina were recruited. Messenger RNA (mRNA) expression of Vα24-Jα18 (invariant TCRα chain of human iNKT cells), IFNγ and intracellular transcription factor Forkhead Box P3 (Foxp3), and flow cytometry intraepithelial lymphocyte (IEL) profile were determined. Both uCD and GFD-CD patients had higher Vα24-Jα18 mRNA levels than non-CD controls (I and C-controls). The expression of Vα24-Jα18 correlated with Marsh score for the severity of mucosal lesion and also with increased mRNA IFNγ levels. uCD and GFD-CD patients had decreased mRNA expression of FoxP3 but increased expression of Vα24-Jα18, which revealed a CD-like molecular profile. Increased numbers of iNKT cells were confirmed by flow cytometry within the intraepithelial lymphocyte compartment of uCD and GFD-CD patients and correlated with Vα24-Jα18 mRNA expression. In conclusion, we have found an increased number of iNKT cells in the duodenum from both uCD and GFD-CD patients, irrespective of the mucosal status. A CD-like molecular profile, defined by an increased mRNA expression of Vα24-Jα18 together with a decreased expression of FoxP3, may represent a pro-inflammatory signature of the CD duodenum.

Show MeSH
Related in: MedlinePlus