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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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IL-10 induces miRNA that affect proinflammatory mediators.[A] RNA was isolated from mBSA rechallenged LN cells treated with vehicle or IL-10 [20, 10 ng/ml], as described in Fig 4. qPCR was performed to analyze changes in miRNA 21, 504, 125a, 30e, and 301a. Data are representative of 3 individual experiments with n = 3. [B] Ingenuity pathway analysis [IPA] of miRNA targets for miRNA induced by IL-10. * p<0.05 compared to vehicle.
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pone-0093954-g006: IL-10 induces miRNA that affect proinflammatory mediators.[A] RNA was isolated from mBSA rechallenged LN cells treated with vehicle or IL-10 [20, 10 ng/ml], as described in Fig 4. qPCR was performed to analyze changes in miRNA 21, 504, 125a, 30e, and 301a. Data are representative of 3 individual experiments with n = 3. [B] Ingenuity pathway analysis [IPA] of miRNA targets for miRNA induced by IL-10. * p<0.05 compared to vehicle.

Mentions: Because we noted that IL-10 plays a crucial role in the regulation of anti-inflammatory effects of AEA, we next determined if IL-10 treatment induces miRNA that can suppress inflammatory pathways. To this end, we isolated RNA from mBSA sensitized cells treated with vehicle or rIL-10 at 10 and 20 ng/ml and analyzed them for specific miRNA that we identified from our in vivo data which could potentially target inflammatory pathways [Fig 5C]. Interestingly, we found that IL-10 was able to induce the expression of miRNA 30e, 504, 125a, 301a, and 21 [Fig 6A]. It was noteworthy that several of these miRNAs such as 30e, 125a, and 301a had been found to be up-regulated in vivo upon AEA treatment [Fig 4C]. We then used Ingenuity pathway analysis to determine the effect of these miRNAs on pro-inflammatory mediators [Fig 6B]. Interestingly, these microRNAs were found to target several proinflammatory cytokines and their receptors, including IFN-γ, TNF-α, and TGF-β/IL-6 necessary for Th17 differentiation. Along with these, STAT-3 was also targeted by several of these miRNAs. Together, these data suggested that AEA may promote IL-10 induction which in turn may up-regulate certain miRNA that suppress inflammatory pathways.


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)

IL-10 induces miRNA that affect proinflammatory mediators.[A] RNA was isolated from mBSA rechallenged LN cells treated with vehicle or IL-10 [20, 10 ng/ml], as described in Fig 4. qPCR was performed to analyze changes in miRNA 21, 504, 125a, 30e, and 301a. Data are representative of 3 individual experiments with n = 3. [B] Ingenuity pathway analysis [IPA] of miRNA targets for miRNA induced by IL-10. * p<0.05 compared to vehicle.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0093954-g006: IL-10 induces miRNA that affect proinflammatory mediators.[A] RNA was isolated from mBSA rechallenged LN cells treated with vehicle or IL-10 [20, 10 ng/ml], as described in Fig 4. qPCR was performed to analyze changes in miRNA 21, 504, 125a, 30e, and 301a. Data are representative of 3 individual experiments with n = 3. [B] Ingenuity pathway analysis [IPA] of miRNA targets for miRNA induced by IL-10. * p<0.05 compared to vehicle.
Mentions: Because we noted that IL-10 plays a crucial role in the regulation of anti-inflammatory effects of AEA, we next determined if IL-10 treatment induces miRNA that can suppress inflammatory pathways. To this end, we isolated RNA from mBSA sensitized cells treated with vehicle or rIL-10 at 10 and 20 ng/ml and analyzed them for specific miRNA that we identified from our in vivo data which could potentially target inflammatory pathways [Fig 5C]. Interestingly, we found that IL-10 was able to induce the expression of miRNA 30e, 504, 125a, 301a, and 21 [Fig 6A]. It was noteworthy that several of these miRNAs such as 30e, 125a, and 301a had been found to be up-regulated in vivo upon AEA treatment [Fig 4C]. We then used Ingenuity pathway analysis to determine the effect of these miRNAs on pro-inflammatory mediators [Fig 6B]. Interestingly, these microRNAs were found to target several proinflammatory cytokines and their receptors, including IFN-γ, TNF-α, and TGF-β/IL-6 necessary for Th17 differentiation. Along with these, STAT-3 was also targeted by several of these miRNAs. Together, these data suggested that AEA may promote IL-10 induction which in turn may up-regulate certain miRNA that suppress inflammatory pathways.

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