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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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Identification of intraepithelial and lamina propria lymphocytes by flow cytometry. Example of characterization of intraepithelial lymphocytes (IELs)/lamina propria lymphocytes (LPL) in an untreated celiac disease donor. Lamina propria (CD45+) and IEL (CD45+CD103+) were identified and percentages of TCRαβ cells, TCRγδ cells and non-T cells determined (A). Example of characterization of invariant NKT (iNKT) cells in an untreated celiac disease donor: CD45+CD3+Vα24-Jα18+ cells within the total of CD45+CD3+ cells, iNKT (CD45+CD3+Vα24-Jα18+) phenotype according to the expression of CD4 and/or CD8 within the total of iNKTs (B).
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nutrients-07-05444-f001: Identification of intraepithelial and lamina propria lymphocytes by flow cytometry. Example of characterization of intraepithelial lymphocytes (IELs)/lamina propria lymphocytes (LPL) in an untreated celiac disease donor. Lamina propria (CD45+) and IEL (CD45+CD103+) were identified and percentages of TCRαβ cells, TCRγδ cells and non-T cells determined (A). Example of characterization of invariant NKT (iNKT) cells in an untreated celiac disease donor: CD45+CD3+Vα24-Jα18+ cells within the total of CD45+CD3+ cells, iNKT (CD45+CD3+Vα24-Jα18+) phenotype according to the expression of CD4 and/or CD8 within the total of iNKTs (B).

Mentions: Cells were acquired in a Beckman Coulter FC500 flow cytometer and data processed with Cell BC software (Beckman Coulter, Brea, CA, USA). All IEL and lamina propria lymphocyte (LPL) cells were identified as CD45+ (leukocyte pan-marker) and IELs were also identified as CD103+. Non-T cells (CD3−), TCRγδ cells (CD3+TCRγδ+), TCRαβ cells (CD3+TCRγδ−) (Figure 1A), iNKT cells (CD3+Vα24-Jα18+) (Figure 1B) and Treg cells (CD3+CD4+CD25+FoxP3+ or CD3+CD4+FoxP3+) were identified by flow cytometry within the intraepithelial and the lamina propria compartments. Numbers of cells were expressed as percentages.


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)

Identification of intraepithelial and lamina propria lymphocytes by flow cytometry. Example of characterization of intraepithelial lymphocytes (IELs)/lamina propria lymphocytes (LPL) in an untreated celiac disease donor. Lamina propria (CD45+) and IEL (CD45+CD103+) were identified and percentages of TCRαβ cells, TCRγδ cells and non-T cells determined (A). Example of characterization of invariant NKT (iNKT) cells in an untreated celiac disease donor: CD45+CD3+Vα24-Jα18+ cells within the total of CD45+CD3+ cells, iNKT (CD45+CD3+Vα24-Jα18+) phenotype according to the expression of CD4 and/or CD8 within the total of iNKTs (B).
© Copyright Policy
Related In: Results  -  Collection

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

nutrients-07-05444-f001: Identification of intraepithelial and lamina propria lymphocytes by flow cytometry. Example of characterization of intraepithelial lymphocytes (IELs)/lamina propria lymphocytes (LPL) in an untreated celiac disease donor. Lamina propria (CD45+) and IEL (CD45+CD103+) were identified and percentages of TCRαβ cells, TCRγδ cells and non-T cells determined (A). Example of characterization of invariant NKT (iNKT) cells in an untreated celiac disease donor: CD45+CD3+Vα24-Jα18+ cells within the total of CD45+CD3+ cells, iNKT (CD45+CD3+Vα24-Jα18+) phenotype according to the expression of CD4 and/or CD8 within the total of iNKTs (B).
Mentions: Cells were acquired in a Beckman Coulter FC500 flow cytometer and data processed with Cell BC software (Beckman Coulter, Brea, CA, USA). All IEL and lamina propria lymphocyte (LPL) cells were identified as CD45+ (leukocyte pan-marker) and IELs were also identified as CD103+. Non-T cells (CD3−), TCRγδ cells (CD3+TCRγδ+), TCRαβ cells (CD3+TCRγδ−) (Figure 1A), iNKT cells (CD3+Vα24-Jα18+) (Figure 1B) and Treg cells (CD3+CD4+CD25+FoxP3+ or CD3+CD4+FoxP3+) were identified by flow cytometry within the intraepithelial and the lamina propria compartments. Numbers of cells were expressed as percentages.

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