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A microRNA upregulated in asthma airway T cells promotes TH2 cytokine production.

Simpson LJ, Patel S, Bhakta NR, Choy DF, Brightbill HD, Ren X, Wang Y, Pua HH, Baumjohann D, Montoya MM, Panduro M, Remedios KA, Huang X, Fahy JV, Arron JR, Woodruff PG, Ansel KM - Nat. Immunol. (2014)

Bottom Line: MicroRNAs (miRNAs) exert powerful effects on immunological function by tuning networks of target genes that orchestrate cell activity.Modulating miR-19 activity altered TH2 cytokine production in both human and mouse T cells, and TH2 cell responses were markedly impaired in cells lacking the entire miR-17∼92 cluster. miR-19 promoted TH2 cytokine production and amplified inflammatory signaling by direct targeting of the inositol phosphatase PTEN, the signaling inhibitor SOCS1 and the deubiquitinase A20.Thus, upregulation of miR-19a in asthma may be an indicator and a cause of increased TH2 cytokine production in the airways.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Sandler Asthma Basic Research Center, University of California San Francisco, San Francisco, California, USA.

ABSTRACT
MicroRNAs (miRNAs) exert powerful effects on immunological function by tuning networks of target genes that orchestrate cell activity. We sought to identify miRNAs and miRNA-regulated pathways that control the type 2 helper T cell (TH2 cell) responses that drive pathogenic inflammation in asthma. Profiling miRNA expression in human airway-infiltrating T cells revealed elevated expression of the miRNA miR-19a in asthma. Modulating miR-19 activity altered TH2 cytokine production in both human and mouse T cells, and TH2 cell responses were markedly impaired in cells lacking the entire miR-17∼92 cluster. miR-19 promoted TH2 cytokine production and amplified inflammatory signaling by direct targeting of the inositol phosphatase PTEN, the signaling inhibitor SOCS1 and the deubiquitinase A20. Thus, upregulation of miR-19a in asthma may be an indicator and a cause of increased TH2 cytokine production in the airways.

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miR-19a and miR-19b rescue the TH2 cytokine defect in 17∼92Δ/Δ cells(a) Intracellular cytokine staining of 17∼92+/+ or 17∼92Δ/Δ CD4 T cells transfected with control mimic (CM), miR-19a, -19b, -17, -18a, -20a, or -92a mimics. Cells were transfected on days 1 and 4 of TH2 polarization, and analyzed on day 5. Numbers in each quadrant indicate percentage of cells producing the indicated cytokine. Data are representative of 3 independent experiments. (b-c) Quantification of IL-13 (b) or IL-4 (c) production at day 5 after transfection with miRNA mimics. Bars represent mean ± SEM for 3 individual transfections for each condition. Data are representative of 3 independent experiments. (d-e) 17∼92+/+ or 17∼92Δ/Δ CD4+ T cells were transfected with CM, miR-19a, or -19b on day 1 only, day 4 only, or both day 1 and 4. Quantification of IL-13 (d) or IL-4 (e) production at day 5 of TH2 polarization. Error bars are mean ± SEM for 3 individual transfections for each condition. Data are representative of 2 independent experiments. (b, c) One-way ANOVA with Dunnett's post test (comparing each column to 17∼92Δ/Δ + CM). (d, e) p< 0.0001 by 2-way ANOVA with Bonferroni post test (comparing each column to 17∼92Δ/Δ + CM per time point). ns = “not significant”, *p< 0.05, ** p < 0.01, *** p < 0.001, ****p < 0.0001.
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Figure 4: miR-19a and miR-19b rescue the TH2 cytokine defect in 17∼92Δ/Δ cells(a) Intracellular cytokine staining of 17∼92+/+ or 17∼92Δ/Δ CD4 T cells transfected with control mimic (CM), miR-19a, -19b, -17, -18a, -20a, or -92a mimics. Cells were transfected on days 1 and 4 of TH2 polarization, and analyzed on day 5. Numbers in each quadrant indicate percentage of cells producing the indicated cytokine. Data are representative of 3 independent experiments. (b-c) Quantification of IL-13 (b) or IL-4 (c) production at day 5 after transfection with miRNA mimics. Bars represent mean ± SEM for 3 individual transfections for each condition. Data are representative of 3 independent experiments. (d-e) 17∼92+/+ or 17∼92Δ/Δ CD4+ T cells were transfected with CM, miR-19a, or -19b on day 1 only, day 4 only, or both day 1 and 4. Quantification of IL-13 (d) or IL-4 (e) production at day 5 of TH2 polarization. Error bars are mean ± SEM for 3 individual transfections for each condition. Data are representative of 2 independent experiments. (b, c) One-way ANOVA with Dunnett's post test (comparing each column to 17∼92Δ/Δ + CM). (d, e) p< 0.0001 by 2-way ANOVA with Bonferroni post test (comparing each column to 17∼92Δ/Δ + CM per time point). ns = “not significant”, *p< 0.05, ** p < 0.01, *** p < 0.001, ****p < 0.0001.

Mentions: To understand the mechanism of miR-17∼92 control of TH2 cell cytokine production, the functions of individual miRNA members of the cluster need to be addressed. Therefore, we transfected mature miRNA mimics corresponding to each of the six miRNAs in the miR-17∼92 cluster into 17∼92Δ/Δ CD4+ T cells on days 1 and 4 of TH2 polarizing cultures. Transfection of either miR-19a or miR-19b was sufficient to completely rescue TH2 cytokine production (Fig. 4a-c). Other miRNAs within the cluster conferred only partial rescue of TH2 cytokine production compared to miR-19a or miR-19b. These data indicate that miR-19 is the primary component of the miR-17∼92 cluster that augments TH2 differentiation.


A microRNA upregulated in asthma airway T cells promotes TH2 cytokine production.

Simpson LJ, Patel S, Bhakta NR, Choy DF, Brightbill HD, Ren X, Wang Y, Pua HH, Baumjohann D, Montoya MM, Panduro M, Remedios KA, Huang X, Fahy JV, Arron JR, Woodruff PG, Ansel KM - Nat. Immunol. (2014)

miR-19a and miR-19b rescue the TH2 cytokine defect in 17∼92Δ/Δ cells(a) Intracellular cytokine staining of 17∼92+/+ or 17∼92Δ/Δ CD4 T cells transfected with control mimic (CM), miR-19a, -19b, -17, -18a, -20a, or -92a mimics. Cells were transfected on days 1 and 4 of TH2 polarization, and analyzed on day 5. Numbers in each quadrant indicate percentage of cells producing the indicated cytokine. Data are representative of 3 independent experiments. (b-c) Quantification of IL-13 (b) or IL-4 (c) production at day 5 after transfection with miRNA mimics. Bars represent mean ± SEM for 3 individual transfections for each condition. Data are representative of 3 independent experiments. (d-e) 17∼92+/+ or 17∼92Δ/Δ CD4+ T cells were transfected with CM, miR-19a, or -19b on day 1 only, day 4 only, or both day 1 and 4. Quantification of IL-13 (d) or IL-4 (e) production at day 5 of TH2 polarization. Error bars are mean ± SEM for 3 individual transfections for each condition. Data are representative of 2 independent experiments. (b, c) One-way ANOVA with Dunnett's post test (comparing each column to 17∼92Δ/Δ + CM). (d, e) p< 0.0001 by 2-way ANOVA with Bonferroni post test (comparing each column to 17∼92Δ/Δ + CM per time point). ns = “not significant”, *p< 0.05, ** p < 0.01, *** p < 0.001, ****p < 0.0001.
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Figure 4: miR-19a and miR-19b rescue the TH2 cytokine defect in 17∼92Δ/Δ cells(a) Intracellular cytokine staining of 17∼92+/+ or 17∼92Δ/Δ CD4 T cells transfected with control mimic (CM), miR-19a, -19b, -17, -18a, -20a, or -92a mimics. Cells were transfected on days 1 and 4 of TH2 polarization, and analyzed on day 5. Numbers in each quadrant indicate percentage of cells producing the indicated cytokine. Data are representative of 3 independent experiments. (b-c) Quantification of IL-13 (b) or IL-4 (c) production at day 5 after transfection with miRNA mimics. Bars represent mean ± SEM for 3 individual transfections for each condition. Data are representative of 3 independent experiments. (d-e) 17∼92+/+ or 17∼92Δ/Δ CD4+ T cells were transfected with CM, miR-19a, or -19b on day 1 only, day 4 only, or both day 1 and 4. Quantification of IL-13 (d) or IL-4 (e) production at day 5 of TH2 polarization. Error bars are mean ± SEM for 3 individual transfections for each condition. Data are representative of 2 independent experiments. (b, c) One-way ANOVA with Dunnett's post test (comparing each column to 17∼92Δ/Δ + CM). (d, e) p< 0.0001 by 2-way ANOVA with Bonferroni post test (comparing each column to 17∼92Δ/Δ + CM per time point). ns = “not significant”, *p< 0.05, ** p < 0.01, *** p < 0.001, ****p < 0.0001.
Mentions: To understand the mechanism of miR-17∼92 control of TH2 cell cytokine production, the functions of individual miRNA members of the cluster need to be addressed. Therefore, we transfected mature miRNA mimics corresponding to each of the six miRNAs in the miR-17∼92 cluster into 17∼92Δ/Δ CD4+ T cells on days 1 and 4 of TH2 polarizing cultures. Transfection of either miR-19a or miR-19b was sufficient to completely rescue TH2 cytokine production (Fig. 4a-c). Other miRNAs within the cluster conferred only partial rescue of TH2 cytokine production compared to miR-19a or miR-19b. These data indicate that miR-19 is the primary component of the miR-17∼92 cluster that augments TH2 differentiation.

Bottom Line: MicroRNAs (miRNAs) exert powerful effects on immunological function by tuning networks of target genes that orchestrate cell activity.Modulating miR-19 activity altered TH2 cytokine production in both human and mouse T cells, and TH2 cell responses were markedly impaired in cells lacking the entire miR-17∼92 cluster. miR-19 promoted TH2 cytokine production and amplified inflammatory signaling by direct targeting of the inositol phosphatase PTEN, the signaling inhibitor SOCS1 and the deubiquitinase A20.Thus, upregulation of miR-19a in asthma may be an indicator and a cause of increased TH2 cytokine production in the airways.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Sandler Asthma Basic Research Center, University of California San Francisco, San Francisco, California, USA.

ABSTRACT
MicroRNAs (miRNAs) exert powerful effects on immunological function by tuning networks of target genes that orchestrate cell activity. We sought to identify miRNAs and miRNA-regulated pathways that control the type 2 helper T cell (TH2 cell) responses that drive pathogenic inflammation in asthma. Profiling miRNA expression in human airway-infiltrating T cells revealed elevated expression of the miRNA miR-19a in asthma. Modulating miR-19 activity altered TH2 cytokine production in both human and mouse T cells, and TH2 cell responses were markedly impaired in cells lacking the entire miR-17∼92 cluster. miR-19 promoted TH2 cytokine production and amplified inflammatory signaling by direct targeting of the inositol phosphatase PTEN, the signaling inhibitor SOCS1 and the deubiquitinase A20. Thus, upregulation of miR-19a in asthma may be an indicator and a cause of increased TH2 cytokine production in the airways.

Show MeSH
Related in: MedlinePlus