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Human gastric epithelial cells contribute to gastric immune regulation by providing retinoic acid to dendritic cells.

Bimczok D, Kao JY, Zhang M, Cochrun S, Mannon P, Peter S, Wilcox CM, Mönkemüller KE, Harris PR, Grams JM, Stahl RD, Smith PD, Smythies LE - Mucosal Immunol (2014)

Bottom Line: Indeed, DCs purified from gastric mucosa had similar levels of aldehyde dehydrogenase activity and RA biosynthesis gene expression as small intestinal DCs, although gastric DCs lacked CD103.In H. pylori-infected gastric mucosa, gastric RA biosynthesis gene expression was severely disrupted, which may lead to reduced RA signaling and thus contribute to disease progression.Collectively, our results support a critical role for RA in human gastric immune regulation.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA.

ABSTRACT
Despite the high prevalence of chronic gastritis caused by Helicobacter pylori, the gastric mucosa has received little investigative attention as a unique immune environment. Here, we analyzed whether retinoic acid (RA), an important homeostatic factor in the small intestinal mucosa, also contributes to gastric immune regulation. We report that human gastric tissue contains high levels of the RA precursor molecule retinol (ROL), and that gastric epithelial cells express both RA biosynthesis genes and RA response genes, indicative of active RA biosynthesis. Moreover, primary gastric epithelial cells cultured in the presence of ROL synthesized RA in vitro and induced RA biosynthesis in co-cultured monocytes through an RA-dependent mechanism, suggesting that gastric epithelial cells may also confer the ability to generate RA on gastric dendritic cells (DCs). Indeed, DCs purified from gastric mucosa had similar levels of aldehyde dehydrogenase activity and RA biosynthesis gene expression as small intestinal DCs, although gastric DCs lacked CD103. In H. pylori-infected gastric mucosa, gastric RA biosynthesis gene expression was severely disrupted, which may lead to reduced RA signaling and thus contribute to disease progression. Collectively, our results support a critical role for RA in human gastric immune regulation.

No MeSH data available.


Related in: MedlinePlus

Gastric DCs lack expression of CD103(a) Gating strategy for isolation of gastric and intestinal DCs. (b) Phenotype of isolated gastric DCs. Data are representative of n=6–8. (c) Intestinal DCs express higher levels of CD103 than gastric DCs. Diamonds: individual samples; bars: mean; P=0.02, Kruskal-Wallis test with Bonferroni correction; n=6–8. (d, e) Gastric DCs lack ITGAE (CD103) gene expression but express similar levels of ZBTB46. RNA was isolated from FACS-sorted gastric and intestinal dendritic cells (live HLA-DR+/CD45+/lineage− cells), and expression of (d) ITGAE and (e) ZBTB46 was analyzed by qRT-PCR. Monocyte-derived DCs (MoDCs; n=1) and elutriated intestinal macrophages (Macs, n=1) were analyzed for comparison. Diamonds: individual samples; bars: mean; lines connect paired samples; n=7. Statistical significance was determined using the Student’s t-test.
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Figure 5: Gastric DCs lack expression of CD103(a) Gating strategy for isolation of gastric and intestinal DCs. (b) Phenotype of isolated gastric DCs. Data are representative of n=6–8. (c) Intestinal DCs express higher levels of CD103 than gastric DCs. Diamonds: individual samples; bars: mean; P=0.02, Kruskal-Wallis test with Bonferroni correction; n=6–8. (d, e) Gastric DCs lack ITGAE (CD103) gene expression but express similar levels of ZBTB46. RNA was isolated from FACS-sorted gastric and intestinal dendritic cells (live HLA-DR+/CD45+/lineage− cells), and expression of (d) ITGAE and (e) ZBTB46 was analyzed by qRT-PCR. Monocyte-derived DCs (MoDCs; n=1) and elutriated intestinal macrophages (Macs, n=1) were analyzed for comparison. Diamonds: individual samples; bars: mean; lines connect paired samples; n=7. Statistical significance was determined using the Student’s t-test.

Mentions: CD103+ DCs, the major DC subset in murine small intestinal lamina propria, have an enhanced capacity to generate RA (6, 25), but whether human gastric DCs express CD103 and synthesize RA is currently unknown. We have previously characterized human gastric DCs as HLA-DRhigh/CD13low cells that induce CD4+ T cell IFN-γ production but only weak proliferative responses (2, 26). Here, we show that freshly isolated human gastric DCs gated as live/lineage/HLA-DR high/CD45+ singlet cells (Fig. 5a) showed subset expression of CD11c (46±11%, n=6), BDCA1 (CD1c, 28±9%), DC-SIGN (CD209, 13±3%) and SIRP-α (CD172a, 27±10%), but expressed only low levels of CXCR1 (4±1%) and CD103 (3±1%) (Fig. 5b). In contrast, similarly gated DCs from human small intestine expressed significantly higher levels of CD103 protein (Fig. 5c, P=0.02, Kruskal-Wallis test with Bonferroni correction). The observed difference in CD103 expression between gastric and intestinal DCs was corroborated by quantitative RT-PCR analysis, which also revealed a significantly lower expression of ITGAE (CD103) mRNA in gastric compared with intestinal DCs (Fig. 5d, P=0.002). Importantly, FACS-purified gastric and intestinal DCs expressed similar levels of the DC-specific transcription factor ZBTB46, confirming that both cell populations both were DCs (27) (Fig. 5e). In comparison, control MoDCs displayed high expression of ITGAE and low expression of ZBTB46, whereas intestinal macrophages showed low expression of both CD103 and ZBTB46.


Human gastric epithelial cells contribute to gastric immune regulation by providing retinoic acid to dendritic cells.

Bimczok D, Kao JY, Zhang M, Cochrun S, Mannon P, Peter S, Wilcox CM, Mönkemüller KE, Harris PR, Grams JM, Stahl RD, Smith PD, Smythies LE - Mucosal Immunol (2014)

Gastric DCs lack expression of CD103(a) Gating strategy for isolation of gastric and intestinal DCs. (b) Phenotype of isolated gastric DCs. Data are representative of n=6–8. (c) Intestinal DCs express higher levels of CD103 than gastric DCs. Diamonds: individual samples; bars: mean; P=0.02, Kruskal-Wallis test with Bonferroni correction; n=6–8. (d, e) Gastric DCs lack ITGAE (CD103) gene expression but express similar levels of ZBTB46. RNA was isolated from FACS-sorted gastric and intestinal dendritic cells (live HLA-DR+/CD45+/lineage− cells), and expression of (d) ITGAE and (e) ZBTB46 was analyzed by qRT-PCR. Monocyte-derived DCs (MoDCs; n=1) and elutriated intestinal macrophages (Macs, n=1) were analyzed for comparison. Diamonds: individual samples; bars: mean; lines connect paired samples; n=7. Statistical significance was determined using the Student’s t-test.
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Figure 5: Gastric DCs lack expression of CD103(a) Gating strategy for isolation of gastric and intestinal DCs. (b) Phenotype of isolated gastric DCs. Data are representative of n=6–8. (c) Intestinal DCs express higher levels of CD103 than gastric DCs. Diamonds: individual samples; bars: mean; P=0.02, Kruskal-Wallis test with Bonferroni correction; n=6–8. (d, e) Gastric DCs lack ITGAE (CD103) gene expression but express similar levels of ZBTB46. RNA was isolated from FACS-sorted gastric and intestinal dendritic cells (live HLA-DR+/CD45+/lineage− cells), and expression of (d) ITGAE and (e) ZBTB46 was analyzed by qRT-PCR. Monocyte-derived DCs (MoDCs; n=1) and elutriated intestinal macrophages (Macs, n=1) were analyzed for comparison. Diamonds: individual samples; bars: mean; lines connect paired samples; n=7. Statistical significance was determined using the Student’s t-test.
Mentions: CD103+ DCs, the major DC subset in murine small intestinal lamina propria, have an enhanced capacity to generate RA (6, 25), but whether human gastric DCs express CD103 and synthesize RA is currently unknown. We have previously characterized human gastric DCs as HLA-DRhigh/CD13low cells that induce CD4+ T cell IFN-γ production but only weak proliferative responses (2, 26). Here, we show that freshly isolated human gastric DCs gated as live/lineage/HLA-DR high/CD45+ singlet cells (Fig. 5a) showed subset expression of CD11c (46±11%, n=6), BDCA1 (CD1c, 28±9%), DC-SIGN (CD209, 13±3%) and SIRP-α (CD172a, 27±10%), but expressed only low levels of CXCR1 (4±1%) and CD103 (3±1%) (Fig. 5b). In contrast, similarly gated DCs from human small intestine expressed significantly higher levels of CD103 protein (Fig. 5c, P=0.02, Kruskal-Wallis test with Bonferroni correction). The observed difference in CD103 expression between gastric and intestinal DCs was corroborated by quantitative RT-PCR analysis, which also revealed a significantly lower expression of ITGAE (CD103) mRNA in gastric compared with intestinal DCs (Fig. 5d, P=0.002). Importantly, FACS-purified gastric and intestinal DCs expressed similar levels of the DC-specific transcription factor ZBTB46, confirming that both cell populations both were DCs (27) (Fig. 5e). In comparison, control MoDCs displayed high expression of ITGAE and low expression of ZBTB46, whereas intestinal macrophages showed low expression of both CD103 and ZBTB46.

Bottom Line: Indeed, DCs purified from gastric mucosa had similar levels of aldehyde dehydrogenase activity and RA biosynthesis gene expression as small intestinal DCs, although gastric DCs lacked CD103.In H. pylori-infected gastric mucosa, gastric RA biosynthesis gene expression was severely disrupted, which may lead to reduced RA signaling and thus contribute to disease progression.Collectively, our results support a critical role for RA in human gastric immune regulation.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA.

ABSTRACT
Despite the high prevalence of chronic gastritis caused by Helicobacter pylori, the gastric mucosa has received little investigative attention as a unique immune environment. Here, we analyzed whether retinoic acid (RA), an important homeostatic factor in the small intestinal mucosa, also contributes to gastric immune regulation. We report that human gastric tissue contains high levels of the RA precursor molecule retinol (ROL), and that gastric epithelial cells express both RA biosynthesis genes and RA response genes, indicative of active RA biosynthesis. Moreover, primary gastric epithelial cells cultured in the presence of ROL synthesized RA in vitro and induced RA biosynthesis in co-cultured monocytes through an RA-dependent mechanism, suggesting that gastric epithelial cells may also confer the ability to generate RA on gastric dendritic cells (DCs). Indeed, DCs purified from gastric mucosa had similar levels of aldehyde dehydrogenase activity and RA biosynthesis gene expression as small intestinal DCs, although gastric DCs lacked CD103. In H. pylori-infected gastric mucosa, gastric RA biosynthesis gene expression was severely disrupted, which may lead to reduced RA signaling and thus contribute to disease progression. Collectively, our results support a critical role for RA in human gastric immune regulation.

No MeSH data available.


Related in: MedlinePlus