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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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Phenotype of intraepithelial lymphocytes in the duodenum from celiac patients. Phenotype of IELs 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), analyzed by flow cytometry: Percentage of total IELs (CD103+CD45+) referred to the total of epithelial cells (A). Percentage of non-T cells (CD103+CD45+CD3−) (B), Tγδ cells (CD103+CD45+CD3+TCRγδ+) (C) and Tαβ cells (CD103+CD45+CD3+TCRγδ−) (D) referred to the total of IELs. Horizontal bar are median values. Statistically significant differences are shown (two tailed Mann Whitney U test; p < 0.05). Representative flow cytometry analysis data of IEL subpopulations (Tγδ, Tαβ and non-T cells) in uCD, GFD-CD, I-controls and C-controls (E).
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nutrients-07-05444-f005: Phenotype of intraepithelial lymphocytes in the duodenum from celiac patients. Phenotype of IELs 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), analyzed by flow cytometry: Percentage of total IELs (CD103+CD45+) referred to the total of epithelial cells (A). Percentage of non-T cells (CD103+CD45+CD3−) (B), Tγδ cells (CD103+CD45+CD3+TCRγδ+) (C) and Tαβ cells (CD103+CD45+CD3+TCRγδ−) (D) referred to the total of IELs. Horizontal bar are median values. Statistically significant differences are shown (two tailed Mann Whitney U test; p < 0.05). Representative flow cytometry analysis data of IEL subpopulations (Tγδ, Tαβ and non-T cells) in uCD, GFD-CD, I-controls and C-controls (E).

Mentions: Untreated CD patients had increased numbers of total IELs (median/IQR; 16.80%/4.80) (Figure 5A,E) together with decreased numbers of non-T cells (3.10%/4.00) (Figure 5B,E). The latter was also true in GFD-CD patients although the total number of IELs did not increase (GFD-CD, 10.40%/4.11) (Figure 5A,E). Within the CD3+ subpopulation, both uCD and GFD-CD patients showed higher numbers of TCRγδ+ cells (uCD, 35.78%/11.13; GFD-CD, 32.50%/8.90) (Figure 5C,E), previously described as a distinctive feature of CD patients [9,32]. In support of this, inflamed non-CD controls did not have increased numbers of TCRγδ+ cells (I-controls, 7.51%/2.60) (Figure 5C,E) despite the decreased percentage of non-T cells compared with non-inflamed non-CD controls (I-controls, 16.30%/8.12; C-controls, 28.90%/15.75) (Figure 5B,E). However, these patients had increased numbers of TCRαβ+ cells (I-controls, 76.20%/8.20) compared with the remainder patient groups (uCD, 59.30%/15.6; GFD-CD, 58.80%/10.90; C-controls, 62.10%/14.5) (Figure 5D,E).


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)

Phenotype of intraepithelial lymphocytes in the duodenum from celiac patients. Phenotype of IELs 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), analyzed by flow cytometry: Percentage of total IELs (CD103+CD45+) referred to the total of epithelial cells (A). Percentage of non-T cells (CD103+CD45+CD3−) (B), Tγδ cells (CD103+CD45+CD3+TCRγδ+) (C) and Tαβ cells (CD103+CD45+CD3+TCRγδ−) (D) referred to the total of IELs. Horizontal bar are median values. Statistically significant differences are shown (two tailed Mann Whitney U test; p < 0.05). Representative flow cytometry analysis data of IEL subpopulations (Tγδ, Tαβ and non-T cells) in uCD, GFD-CD, I-controls and C-controls (E).
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nutrients-07-05444-f005: Phenotype of intraepithelial lymphocytes in the duodenum from celiac patients. Phenotype of IELs 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), analyzed by flow cytometry: Percentage of total IELs (CD103+CD45+) referred to the total of epithelial cells (A). Percentage of non-T cells (CD103+CD45+CD3−) (B), Tγδ cells (CD103+CD45+CD3+TCRγδ+) (C) and Tαβ cells (CD103+CD45+CD3+TCRγδ−) (D) referred to the total of IELs. Horizontal bar are median values. Statistically significant differences are shown (two tailed Mann Whitney U test; p < 0.05). Representative flow cytometry analysis data of IEL subpopulations (Tγδ, Tαβ and non-T cells) in uCD, GFD-CD, I-controls and C-controls (E).
Mentions: Untreated CD patients had increased numbers of total IELs (median/IQR; 16.80%/4.80) (Figure 5A,E) together with decreased numbers of non-T cells (3.10%/4.00) (Figure 5B,E). The latter was also true in GFD-CD patients although the total number of IELs did not increase (GFD-CD, 10.40%/4.11) (Figure 5A,E). Within the CD3+ subpopulation, both uCD and GFD-CD patients showed higher numbers of TCRγδ+ cells (uCD, 35.78%/11.13; GFD-CD, 32.50%/8.90) (Figure 5C,E), previously described as a distinctive feature of CD patients [9,32]. In support of this, inflamed non-CD controls did not have increased numbers of TCRγδ+ cells (I-controls, 7.51%/2.60) (Figure 5C,E) despite the decreased percentage of non-T cells compared with non-inflamed non-CD controls (I-controls, 16.30%/8.12; C-controls, 28.90%/15.75) (Figure 5B,E). However, these patients had increased numbers of TCRαβ+ cells (I-controls, 76.20%/8.20) compared with the remainder patient groups (uCD, 59.30%/15.6; GFD-CD, 58.80%/10.90; C-controls, 62.10%/14.5) (Figure 5D,E).

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