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The retinoic acid-metabolizing enzyme Cyp26b1 regulates CD4 T cell differentiation and function.

Chenery A, Burrows K, Antignano F, Underhill TM, Petkovich M, Zaph C - PLoS ONE (2013)

Bottom Line: We examined the role of Cyp26b1, the T cell-specific family member, in CD4(+) T cells.Transfer of naïve Cyp26b1 (-/-) CD4(+) T cells into Rag1 (-/-) mice resulted in significantly reduced disease in a model of T cell-dependent colitis.Our results show that T cell-specific expression of Cyp26b1 is required for the development of T cell-mediated colitis and may be applicable to the development of therapeutics that target Cyp26b1 for the treatment of inflammatory bowel disease.

View Article: PubMed Central - PubMed

Affiliation: The Biomedical Research Centre, University of British Columbia, Vancouver, British Columbia, Canada.

ABSTRACT
The vitamin A metabolite retinoic acid (RA) has potent immunomodulatory properties that affect T cell differentiation, migration and function. However, the precise role of RA metabolism in T cells remains unclear. Catabolism of RA is mediated by the Cyp26 family of cytochrome P450 oxidases. We examined the role of Cyp26b1, the T cell-specific family member, in CD4(+) T cells. Mice with a conditional knockout of Cyp26b1 in T cells (Cyp26b1 (-/-) mice) displayed normal lymphoid development but showed an increased sensitivity to serum retinoids, which led to increased differentiation under both inducible regulatory T (iTreg) cell- and TH17 cell-polarizing conditions in vitro. Further, Cyp26b1 expression was differentially regulated in iTreg and TH17 cells. Transfer of naïve Cyp26b1 (-/-) CD4(+) T cells into Rag1 (-/-) mice resulted in significantly reduced disease in a model of T cell-dependent colitis. Our results show that T cell-specific expression of Cyp26b1 is required for the development of T cell-mediated colitis and may be applicable to the development of therapeutics that target Cyp26b1 for the treatment of inflammatory bowel disease.

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Cyp26b1 limits iTreg and TH17 cell differentiation in vitro.CD4+ T cells were isolated from Cyp26b1fl/fl and Cyp26b1−/− mice and cultured in iTreg cell- or TH17 cell-promoting conditions, in either serum-containing media or in serum-free media. (A) Gene expression of Cyp26b1 (normalized relative to Actb) was measured by qRT-PCR. (B) Frequencies of IL-17a+ TH17 cells and (C) Foxp3+ CD25+ iTreg cells were determined by flow cytometry. Data in (A) represent mean±SEM of 4 independent experiments; Data in (B) and (C) are from one representative experiment of 4 independent experiments.
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pone-0072308-g003: Cyp26b1 limits iTreg and TH17 cell differentiation in vitro.CD4+ T cells were isolated from Cyp26b1fl/fl and Cyp26b1−/− mice and cultured in iTreg cell- or TH17 cell-promoting conditions, in either serum-containing media or in serum-free media. (A) Gene expression of Cyp26b1 (normalized relative to Actb) was measured by qRT-PCR. (B) Frequencies of IL-17a+ TH17 cells and (C) Foxp3+ CD25+ iTreg cells were determined by flow cytometry. Data in (A) represent mean±SEM of 4 independent experiments; Data in (B) and (C) are from one representative experiment of 4 independent experiments.

Mentions: RA plays an important role in the differentiation of naive CD4+ T cells into iTreg and TH17 cells [6], [7], [16]. To directly test whether Cyp26b1-dependent regulation of RA signaling controls iTreg or TH17 cell differentiation, we stimulated CD4+ T cells from Cyp26b1fl/fl and Cyp26b1−/− mice under iTreg cell- and TH17 cell-promoting conditions. Increased expression of Cyp26b1 was observed in TH17 cells, with a lower expression in iTreg cells (Figure 3A). Following stimulation under TH17 cell-promoting conditions, we observed a marked increased frequency of IL-17a-producing CD4+ T cells in the absence of Cyp26b1 (Figure 3B). These results are consistent with the expression pattern of Cyp26b1 and suggest that induction of Cyp26b1 is required for limiting TH17 cell differentiation. Surprisingly, we also found that the absence of Cyp26b1 resulted in heightened frequencies of CD4+CD25+Foxp3+ iTreg cells (Figure 3C), despite the low levels of Cyp26b1 expression observed in iTreg cells. These results suggest that metabolism of RA is important for limiting iTreg and TH17 cell responses. However, we had not added any exogenous RA to these cultures, suggesting that low levels of serum retinoids affect iTreg and TH17 cell differentiation in the absence of Cyp26b1. To directly test this, we repeated the experiment in serum-free media. Under these conditions, we found equivalent frequencies of iTreg cells and TH17 cells following stimulation of CD4+ T cells from both Cyp26b1fl/fl and Cyp26b1−/− mice. Thus, Cyp26b1 regulates RA signaling in T cells and is critical for limiting iTreg and TH17 cell differentiation. Further, these results suggest that physiological levels of RA in naïve cells impacts effector T cell differentiation.


The retinoic acid-metabolizing enzyme Cyp26b1 regulates CD4 T cell differentiation and function.

Chenery A, Burrows K, Antignano F, Underhill TM, Petkovich M, Zaph C - PLoS ONE (2013)

Cyp26b1 limits iTreg and TH17 cell differentiation in vitro.CD4+ T cells were isolated from Cyp26b1fl/fl and Cyp26b1−/− mice and cultured in iTreg cell- or TH17 cell-promoting conditions, in either serum-containing media or in serum-free media. (A) Gene expression of Cyp26b1 (normalized relative to Actb) was measured by qRT-PCR. (B) Frequencies of IL-17a+ TH17 cells and (C) Foxp3+ CD25+ iTreg cells were determined by flow cytometry. Data in (A) represent mean±SEM of 4 independent experiments; Data in (B) and (C) are from one representative experiment of 4 independent experiments.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3750006&req=5

pone-0072308-g003: Cyp26b1 limits iTreg and TH17 cell differentiation in vitro.CD4+ T cells were isolated from Cyp26b1fl/fl and Cyp26b1−/− mice and cultured in iTreg cell- or TH17 cell-promoting conditions, in either serum-containing media or in serum-free media. (A) Gene expression of Cyp26b1 (normalized relative to Actb) was measured by qRT-PCR. (B) Frequencies of IL-17a+ TH17 cells and (C) Foxp3+ CD25+ iTreg cells were determined by flow cytometry. Data in (A) represent mean±SEM of 4 independent experiments; Data in (B) and (C) are from one representative experiment of 4 independent experiments.
Mentions: RA plays an important role in the differentiation of naive CD4+ T cells into iTreg and TH17 cells [6], [7], [16]. To directly test whether Cyp26b1-dependent regulation of RA signaling controls iTreg or TH17 cell differentiation, we stimulated CD4+ T cells from Cyp26b1fl/fl and Cyp26b1−/− mice under iTreg cell- and TH17 cell-promoting conditions. Increased expression of Cyp26b1 was observed in TH17 cells, with a lower expression in iTreg cells (Figure 3A). Following stimulation under TH17 cell-promoting conditions, we observed a marked increased frequency of IL-17a-producing CD4+ T cells in the absence of Cyp26b1 (Figure 3B). These results are consistent with the expression pattern of Cyp26b1 and suggest that induction of Cyp26b1 is required for limiting TH17 cell differentiation. Surprisingly, we also found that the absence of Cyp26b1 resulted in heightened frequencies of CD4+CD25+Foxp3+ iTreg cells (Figure 3C), despite the low levels of Cyp26b1 expression observed in iTreg cells. These results suggest that metabolism of RA is important for limiting iTreg and TH17 cell responses. However, we had not added any exogenous RA to these cultures, suggesting that low levels of serum retinoids affect iTreg and TH17 cell differentiation in the absence of Cyp26b1. To directly test this, we repeated the experiment in serum-free media. Under these conditions, we found equivalent frequencies of iTreg cells and TH17 cells following stimulation of CD4+ T cells from both Cyp26b1fl/fl and Cyp26b1−/− mice. Thus, Cyp26b1 regulates RA signaling in T cells and is critical for limiting iTreg and TH17 cell differentiation. Further, these results suggest that physiological levels of RA in naïve cells impacts effector T cell differentiation.

Bottom Line: We examined the role of Cyp26b1, the T cell-specific family member, in CD4(+) T cells.Transfer of naïve Cyp26b1 (-/-) CD4(+) T cells into Rag1 (-/-) mice resulted in significantly reduced disease in a model of T cell-dependent colitis.Our results show that T cell-specific expression of Cyp26b1 is required for the development of T cell-mediated colitis and may be applicable to the development of therapeutics that target Cyp26b1 for the treatment of inflammatory bowel disease.

View Article: PubMed Central - PubMed

Affiliation: The Biomedical Research Centre, University of British Columbia, Vancouver, British Columbia, Canada.

ABSTRACT
The vitamin A metabolite retinoic acid (RA) has potent immunomodulatory properties that affect T cell differentiation, migration and function. However, the precise role of RA metabolism in T cells remains unclear. Catabolism of RA is mediated by the Cyp26 family of cytochrome P450 oxidases. We examined the role of Cyp26b1, the T cell-specific family member, in CD4(+) T cells. Mice with a conditional knockout of Cyp26b1 in T cells (Cyp26b1 (-/-) mice) displayed normal lymphoid development but showed an increased sensitivity to serum retinoids, which led to increased differentiation under both inducible regulatory T (iTreg) cell- and TH17 cell-polarizing conditions in vitro. Further, Cyp26b1 expression was differentially regulated in iTreg and TH17 cells. Transfer of naïve Cyp26b1 (-/-) CD4(+) T cells into Rag1 (-/-) mice resulted in significantly reduced disease in a model of T cell-dependent colitis. Our results show that T cell-specific expression of Cyp26b1 is required for the development of T cell-mediated colitis and may be applicable to the development of therapeutics that target Cyp26b1 for the treatment of inflammatory bowel disease.

Show MeSH
Related in: MedlinePlus