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Expression of miRNAs miR-133b and miR-206 in the Il17a/f locus is co-regulated with IL-17 production in αβ and γδ T cells.

Haas JD, Nistala K, Petermann F, Saran N, Chennupati V, Schmitz S, Korn T, Wedderburn LR, Förster R, Krueger A, Prinz I - PLoS ONE (2011)

Bottom Line: Expression of these microRNAs was specific for Th17 as compared to other CD4(+) T cell subsets and this was equally valid for in vitro polarized and ex vivo derived cells.Importantly, we found a similar co-regulation in CCR6(+) and other γδ T cell subsets that are predisposed to production of IL-17.This qualifies expression of miR-133b and miR-206 in T cells as novel biomarkers for Th17-type immune reactions.

View Article: PubMed Central - PubMed

Affiliation: Institute for Immunology, Hannover Medical School, Hannover, Germany.

ABSTRACT
Differentiation of T helper 17 cells (Th17) is a multistep process that involves the cytokines IL-6, TGF-β, and IL-23 as well as IL-1β, IL-21, and TNF-α. Thereby, robust induction of the capacity to produce IL-17 involves epigenetic modifications of the syntenic Il17a/f locus. Using inbred mouse strains, we identified co-regulation of gene transcription at the Il17a/f locus with the nearby microRNAs miR-133b and miR-206 that are clustered approximately 45 kb upstream of Il17a/f. Expression of these microRNAs was specific for Th17 as compared to other CD4(+) T cell subsets and this was equally valid for in vitro polarized and ex vivo derived cells. From all factors analyzed, IL-23 was the most important cytokine for the in vitro induction of miR-133b and miR-206 in naive CD4(+) T cells of wild type mice. However, analysis of IL-23R deficient mice revealed that IL-23R signaling was not essential for the induction of miR-133b and miR-206. Importantly, we found a similar co-regulation in CCR6(+) and other γδ T cell subsets that are predisposed to production of IL-17. Taken together, we discovered a novel feature of T cell differentiation towards an IL-17-producing phenotype that is shared between αβ and γδ T cells. Notably, the specific co-regulation of miR-133b and miR-206 with the Il17a/f locus also extended to human Th17 cells. This qualifies expression of miR-133b and miR-206 in T cells as novel biomarkers for Th17-type immune reactions.

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Primary human Th17 cells, but not Th1 cells, express miR-133b and miR-206.CD4+ T cells were isolated from peripheral blood of 3 human healthy donors, enriched using magnetic beads and then FACS sorted into 3 different populations: IL-17A+, IL-17A− IFNγ+ and IL-17A−IFN-γ−. Expression levels of miR-133b and miR-206 were analyzed by qRT-PCR. Values are plotted as fold increase compared to the respective IL-17A− IFN-γ− population. P-value for miR-206 in IL-17A+ compared to IFN-γ+ expressing cells was 0.0645. Error bars show ±SEM for the 3 different donors.
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pone-0020171-g006: Primary human Th17 cells, but not Th1 cells, express miR-133b and miR-206.CD4+ T cells were isolated from peripheral blood of 3 human healthy donors, enriched using magnetic beads and then FACS sorted into 3 different populations: IL-17A+, IL-17A− IFNγ+ and IL-17A−IFN-γ−. Expression levels of miR-133b and miR-206 were analyzed by qRT-PCR. Values are plotted as fold increase compared to the respective IL-17A− IFN-γ− population. P-value for miR-206 in IL-17A+ compared to IFN-γ+ expressing cells was 0.0645. Error bars show ±SEM for the 3 different donors.

Mentions: Human Th17 cells have been suggested to be involved in diseases such as multiple sclerosis [31] or juvenile idiopathic arthritis [32], but have been described to differ in certain aspects from their murine counterparts, e.g. human Th17 cells co-express IFN-γ more often than murine Th17 cells [33]. Given these apparent differences, we tested whether miR-206/133b and IL-17 were co-regulated in human Th17 cells as well. Therefore, we isolated human primary Th17, Th1 and Th0 cells from healthy donors using a combination of IL-17 and IFN-γ cytokine secretion assays as described [34]. Although a certain degree of donor variability was observed, expression levels of miR-133b and miR-206 were consistently higher in IL-17A secreting human T cells (2 to 4-fold and 15 to 60-fold, respectively) when compared to Th0 cells (Fig. 6). In contrast, miR-206/133b levels were largely identical between human Th1 and Th0 cells. Thus, our data indicate that the miR-206/133b cluster is co-regulated with IL-17 in human T cells as well. In conclusion, we provide evidence that the two miRNAs of the miR-206/133b cluster neighboring the Il17a/f locus are co-regulated in murine and human lymphocytes.


Expression of miRNAs miR-133b and miR-206 in the Il17a/f locus is co-regulated with IL-17 production in αβ and γδ T cells.

Haas JD, Nistala K, Petermann F, Saran N, Chennupati V, Schmitz S, Korn T, Wedderburn LR, Förster R, Krueger A, Prinz I - PLoS ONE (2011)

Primary human Th17 cells, but not Th1 cells, express miR-133b and miR-206.CD4+ T cells were isolated from peripheral blood of 3 human healthy donors, enriched using magnetic beads and then FACS sorted into 3 different populations: IL-17A+, IL-17A− IFNγ+ and IL-17A−IFN-γ−. Expression levels of miR-133b and miR-206 were analyzed by qRT-PCR. Values are plotted as fold increase compared to the respective IL-17A− IFN-γ− population. P-value for miR-206 in IL-17A+ compared to IFN-γ+ expressing cells was 0.0645. Error bars show ±SEM for the 3 different donors.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3102691&req=5

pone-0020171-g006: Primary human Th17 cells, but not Th1 cells, express miR-133b and miR-206.CD4+ T cells were isolated from peripheral blood of 3 human healthy donors, enriched using magnetic beads and then FACS sorted into 3 different populations: IL-17A+, IL-17A− IFNγ+ and IL-17A−IFN-γ−. Expression levels of miR-133b and miR-206 were analyzed by qRT-PCR. Values are plotted as fold increase compared to the respective IL-17A− IFN-γ− population. P-value for miR-206 in IL-17A+ compared to IFN-γ+ expressing cells was 0.0645. Error bars show ±SEM for the 3 different donors.
Mentions: Human Th17 cells have been suggested to be involved in diseases such as multiple sclerosis [31] or juvenile idiopathic arthritis [32], but have been described to differ in certain aspects from their murine counterparts, e.g. human Th17 cells co-express IFN-γ more often than murine Th17 cells [33]. Given these apparent differences, we tested whether miR-206/133b and IL-17 were co-regulated in human Th17 cells as well. Therefore, we isolated human primary Th17, Th1 and Th0 cells from healthy donors using a combination of IL-17 and IFN-γ cytokine secretion assays as described [34]. Although a certain degree of donor variability was observed, expression levels of miR-133b and miR-206 were consistently higher in IL-17A secreting human T cells (2 to 4-fold and 15 to 60-fold, respectively) when compared to Th0 cells (Fig. 6). In contrast, miR-206/133b levels were largely identical between human Th1 and Th0 cells. Thus, our data indicate that the miR-206/133b cluster is co-regulated with IL-17 in human T cells as well. In conclusion, we provide evidence that the two miRNAs of the miR-206/133b cluster neighboring the Il17a/f locus are co-regulated in murine and human lymphocytes.

Bottom Line: Expression of these microRNAs was specific for Th17 as compared to other CD4(+) T cell subsets and this was equally valid for in vitro polarized and ex vivo derived cells.Importantly, we found a similar co-regulation in CCR6(+) and other γδ T cell subsets that are predisposed to production of IL-17.This qualifies expression of miR-133b and miR-206 in T cells as novel biomarkers for Th17-type immune reactions.

View Article: PubMed Central - PubMed

Affiliation: Institute for Immunology, Hannover Medical School, Hannover, Germany.

ABSTRACT
Differentiation of T helper 17 cells (Th17) is a multistep process that involves the cytokines IL-6, TGF-β, and IL-23 as well as IL-1β, IL-21, and TNF-α. Thereby, robust induction of the capacity to produce IL-17 involves epigenetic modifications of the syntenic Il17a/f locus. Using inbred mouse strains, we identified co-regulation of gene transcription at the Il17a/f locus with the nearby microRNAs miR-133b and miR-206 that are clustered approximately 45 kb upstream of Il17a/f. Expression of these microRNAs was specific for Th17 as compared to other CD4(+) T cell subsets and this was equally valid for in vitro polarized and ex vivo derived cells. From all factors analyzed, IL-23 was the most important cytokine for the in vitro induction of miR-133b and miR-206 in naive CD4(+) T cells of wild type mice. However, analysis of IL-23R deficient mice revealed that IL-23R signaling was not essential for the induction of miR-133b and miR-206. Importantly, we found a similar co-regulation in CCR6(+) and other γδ T cell subsets that are predisposed to production of IL-17. Taken together, we discovered a novel feature of T cell differentiation towards an IL-17-producing phenotype that is shared between αβ and γδ T cells. Notably, the specific co-regulation of miR-133b and miR-206 with the Il17a/f locus also extended to human Th17 cells. This qualifies expression of miR-133b and miR-206 in T cells as novel biomarkers for Th17-type immune reactions.

Show MeSH
Related in: MedlinePlus