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Anandamide attenuates Th-17 cell-mediated delayed-type hypersensitivity response by triggering IL-10 production and consequent microRNA induction.

Jackson AR, Nagarkatti P, Nagarkatti M - PLoS ONE (2014)

Bottom Line: AEA treatment significantly reduced IL-17 and IFN-γ production, as well as decreased RORγt expression while causing significant induction of IL-10 in the draining LNs.IL-10 was critical for the AEA-induced mitigation of DTH response inasmuch as neutralization of IL-10 reversed the effects of AEA.Together, the current study demonstrates that AEA may suppress Th-17 cell-mediated DTH response by inducing IL-10 which in turn triggers miRNA that target proinflammatory pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Microbiology, and Immunology, University of South Carolina School of Medicine, Columbia, South Carolina, United States of America.

ABSTRACT
Endogenous cannabinoids [endocannabinoids] are lipid signaling molecules that have been shown to modulate immune functions. However, their role in the regulation of Th17 cells has not been studied previously. In the current study, we used methylated Bovine Serum Albumin [mBSA]-induced delayed type hypersensitivity [DTH] response in C57BL/6 mice, mediated by Th17 cells, as a model to test the anti-inflammatory effects of endocannabinoids. Administration of anandamide [AEA], a member of the endocannabinoid family, into mice resulted in significant mitigation of mBSA-induced inflammation, including foot pad swelling, cell infiltration, and cell proliferation in the draining lymph nodes [LN]. AEA treatment significantly reduced IL-17 and IFN-γ production, as well as decreased RORγt expression while causing significant induction of IL-10 in the draining LNs. IL-10 was critical for the AEA-induced mitigation of DTH response inasmuch as neutralization of IL-10 reversed the effects of AEA. We next analyzed miRNA from the LN cells and found that 100 out of 609 miRNA species were differentially regulated in AEA-treated mice when compared to controls. Several of these miRNAs targeted proinflammatory mediators. Interestingly, many of these miRNA were also upregulated upon in vitro treatment of LN cells with IL-10. Together, the current study demonstrates that AEA may suppress Th-17 cell-mediated DTH response by inducing IL-10 which in turn triggers miRNA that target proinflammatory pathways.

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Effect of AEA on miRNA induction in mice with mBSA-induced DTH: Affymetrix miRNA array analysis.[A] Heat map of miRNA expression patterns. RNA was isolated from five pooled popliteal lymph nodes of mBSA-sensitized mice rechallenged with mBSA and treated with either AEA or Vehicle. The samples were analyzed using Affymetrix miRNA 1.0 microarray. The values for each sample were generated by normalizing the expression of the microRNA to each sample. Ward's Method was used for clustering patterns and the samples were ordered by the Input Rank method. [B] AEA induced differential regulation of several miRNA species, as enumerated by the Venn diagram. [C] The most differential regulated miRNA expressed as a bar chart. [D] Ingenuity Pathway Analysis of the most differentially regulated biological networks.
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pone-0093954-g005: Effect of AEA on miRNA induction in mice with mBSA-induced DTH: Affymetrix miRNA array analysis.[A] Heat map of miRNA expression patterns. RNA was isolated from five pooled popliteal lymph nodes of mBSA-sensitized mice rechallenged with mBSA and treated with either AEA or Vehicle. The samples were analyzed using Affymetrix miRNA 1.0 microarray. The values for each sample were generated by normalizing the expression of the microRNA to each sample. Ward's Method was used for clustering patterns and the samples were ordered by the Input Rank method. [B] AEA induced differential regulation of several miRNA species, as enumerated by the Venn diagram. [C] The most differential regulated miRNA expressed as a bar chart. [D] Ingenuity Pathway Analysis of the most differentially regulated biological networks.

Mentions: Recent studies have shown that microRNAs [miRNA] can regulate cytokines and inflammatory diseases [25]-[27]. To test if the immunosuppression by AEA was mediated by miRNA, we isolated total RNA from draining lymph nodes in mice sensitized to mBSA and treated with vehicle or AEA, as before. We analyzed those samples using Affymetrix miRNA 1.0 microarray. We analyzed 609 different miRNA species for changes in AEA treated mice compared to vehicle controls [Fig 5A]. We found that AEA showed a different expression pattern of miRNA species than vehicle-treated mice. Specifically, there were 100 miRNA species that were differentially regulated in AEA-treated animals, 55 upregulated 1.5 fold or greater and 45 downregulated 1.5 fold or greater [Fig 5B]. We next looked for the most differentially regulated miRNA species. We found that AEA highly upregulated the expression of miRNA 125a-5p [+3.35], miRNA 301a [+2.50], miRNA 30e [+2.57], miRNA 151 [+2.32], and many others. Conversely, AEA downregulated miRNA 374 [−2.38], miRNA 491 [−2.46], and miRNA 132 [−2.89], among others [Fig 5C]. Using Ingenuity Pathway Analysis, we found that AEA significantly affected several miRNA-regulated canonical biological pathways [Fig 5D]. Among these, Infection Disease, Cell Cycle, Cell Death and Survival, and the Cell-mediated Immune Response were of interest in our system.


Anandamide attenuates Th-17 cell-mediated delayed-type hypersensitivity response by triggering IL-10 production and consequent microRNA induction.

Jackson AR, Nagarkatti P, Nagarkatti M - PLoS ONE (2014)

Effect of AEA on miRNA induction in mice with mBSA-induced DTH: Affymetrix miRNA array analysis.[A] Heat map of miRNA expression patterns. RNA was isolated from five pooled popliteal lymph nodes of mBSA-sensitized mice rechallenged with mBSA and treated with either AEA or Vehicle. The samples were analyzed using Affymetrix miRNA 1.0 microarray. The values for each sample were generated by normalizing the expression of the microRNA to each sample. Ward's Method was used for clustering patterns and the samples were ordered by the Input Rank method. [B] AEA induced differential regulation of several miRNA species, as enumerated by the Venn diagram. [C] The most differential regulated miRNA expressed as a bar chart. [D] Ingenuity Pathway Analysis of the most differentially regulated biological networks.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3974854&req=5

pone-0093954-g005: Effect of AEA on miRNA induction in mice with mBSA-induced DTH: Affymetrix miRNA array analysis.[A] Heat map of miRNA expression patterns. RNA was isolated from five pooled popliteal lymph nodes of mBSA-sensitized mice rechallenged with mBSA and treated with either AEA or Vehicle. The samples were analyzed using Affymetrix miRNA 1.0 microarray. The values for each sample were generated by normalizing the expression of the microRNA to each sample. Ward's Method was used for clustering patterns and the samples were ordered by the Input Rank method. [B] AEA induced differential regulation of several miRNA species, as enumerated by the Venn diagram. [C] The most differential regulated miRNA expressed as a bar chart. [D] Ingenuity Pathway Analysis of the most differentially regulated biological networks.
Mentions: Recent studies have shown that microRNAs [miRNA] can regulate cytokines and inflammatory diseases [25]-[27]. To test if the immunosuppression by AEA was mediated by miRNA, we isolated total RNA from draining lymph nodes in mice sensitized to mBSA and treated with vehicle or AEA, as before. We analyzed those samples using Affymetrix miRNA 1.0 microarray. We analyzed 609 different miRNA species for changes in AEA treated mice compared to vehicle controls [Fig 5A]. We found that AEA showed a different expression pattern of miRNA species than vehicle-treated mice. Specifically, there were 100 miRNA species that were differentially regulated in AEA-treated animals, 55 upregulated 1.5 fold or greater and 45 downregulated 1.5 fold or greater [Fig 5B]. We next looked for the most differentially regulated miRNA species. We found that AEA highly upregulated the expression of miRNA 125a-5p [+3.35], miRNA 301a [+2.50], miRNA 30e [+2.57], miRNA 151 [+2.32], and many others. Conversely, AEA downregulated miRNA 374 [−2.38], miRNA 491 [−2.46], and miRNA 132 [−2.89], among others [Fig 5C]. Using Ingenuity Pathway Analysis, we found that AEA significantly affected several miRNA-regulated canonical biological pathways [Fig 5D]. Among these, Infection Disease, Cell Cycle, Cell Death and Survival, and the Cell-mediated Immune Response were of interest in our system.

Bottom Line: AEA treatment significantly reduced IL-17 and IFN-γ production, as well as decreased RORγt expression while causing significant induction of IL-10 in the draining LNs.IL-10 was critical for the AEA-induced mitigation of DTH response inasmuch as neutralization of IL-10 reversed the effects of AEA.Together, the current study demonstrates that AEA may suppress Th-17 cell-mediated DTH response by inducing IL-10 which in turn triggers miRNA that target proinflammatory pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Microbiology, and Immunology, University of South Carolina School of Medicine, Columbia, South Carolina, United States of America.

ABSTRACT
Endogenous cannabinoids [endocannabinoids] are lipid signaling molecules that have been shown to modulate immune functions. However, their role in the regulation of Th17 cells has not been studied previously. In the current study, we used methylated Bovine Serum Albumin [mBSA]-induced delayed type hypersensitivity [DTH] response in C57BL/6 mice, mediated by Th17 cells, as a model to test the anti-inflammatory effects of endocannabinoids. Administration of anandamide [AEA], a member of the endocannabinoid family, into mice resulted in significant mitigation of mBSA-induced inflammation, including foot pad swelling, cell infiltration, and cell proliferation in the draining lymph nodes [LN]. AEA treatment significantly reduced IL-17 and IFN-γ production, as well as decreased RORγt expression while causing significant induction of IL-10 in the draining LNs. IL-10 was critical for the AEA-induced mitigation of DTH response inasmuch as neutralization of IL-10 reversed the effects of AEA. We next analyzed miRNA from the LN cells and found that 100 out of 609 miRNA species were differentially regulated in AEA-treated mice when compared to controls. Several of these miRNAs targeted proinflammatory mediators. Interestingly, many of these miRNA were also upregulated upon in vitro treatment of LN cells with IL-10. Together, the current study demonstrates that AEA may suppress Th-17 cell-mediated DTH response by inducing IL-10 which in turn triggers miRNA that target proinflammatory pathways.

Show MeSH
Related in: MedlinePlus