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Journal of Molecular Medicine

, Volume 94, Issue 9, pp 1039–1051 | Cite as

Marijuana-derived Δ-9-tetrahydrocannabinol suppresses Th1/Th17 cell-mediated delayed-type hypersensitivity through microRNA regulation

  • Jessica M. Sido
  • Austin R. Jackson
  • Prakash S. Nagarkatti
  • Mitzi NagarkattiEmail author
Original Article

Abstract

9-Tetrahydrocannabinol (THC) is one of the major bioactive cannabinoids derived from the Cannabis sativa plant and is known for its anti-inflammatory properties. Delayed-type hypersensitivity (DTH) is driven by proinflammatory T helper cells including the classic inflammatory Th1 lineage as well as the more recently discovered Th17 lineage. In the current study, we investigated whether THC can alter the induction of Th1/Th17 cells involved in mBSA-induced DTH response. THC treatment (20 mg/kg) of C57BL/6 mice with DTH caused decreased swelling and infiltration of immune cells at the site of antigen rechallenge. Additionally, THC treatment decreased lymphocyte activation as well as Th1/Th17 lineage commitment, including reduced lineage-specific transcription factors and cytokines. Interestingly, while DTH caused an overexpression of miR-21, which increases Th17 differentiation via SMAD7 inhibition, and downregulation of miR-29b, an IFN-γ inhibitor, THC treatment reversed this microRNA (miR) dysregulation. Furthermore, when we transfected primary cells from DTH mice with miR-21 inhibitor or miR-29b mimic, as seen with THC treatment, the expression of target gene message was directly impacted increasing SMAD7 and decreasing IFN-γ expression, respectively. In summary, the current study suggests that THC treatment during DTH response can simultaneously inhibit Th1/Th17 activation via regulation of microRNA (miRNA) expression.

Key messages

• THC treatment inhibits simultaneous Th1/Th17 driven inflammation.

• THC treatment corrects DTH-mediated microRNA dysregulation.

• THC treatment regulates proinflammatory cytokines and transcription factors.

Keywords

Hypersensitivity microRNA Cannabinoid THC Inflammation Th1/Th17 regulation 

Notes

Acknowledgments

We wish to acknowledge the University of South Carolina School of Medicine Instrumentation Resource Facility (IRF) for their assistance with histology processing. This work was supported in part by the National Institute of Health Grants R01 MH094755, P01 AT003961 and P20 GM103641 to PN, R01 ES019313 to PN and MN, and R01 AT006888 and the Veterans Administration Merit Award BX001357 to MN.

Compliance with ethical standards

All experiments were conducted under an approved Institutional Animal Care and Use Committee animal protocol. This model of footpad induced DTH is well established and approved by IACUC (protocol# 2214-100895-090814: Mouse Models of Innate Immunity 1031 of the series Methods in Molecular Biology 101-107). The manuscript does not contain clinical studies or patient data.

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

109_2016_1404_MOESM1_ESM.pdf (293 kb)
Supplementary Fig. 1 THC treatment reduces activated lymphocyte proportions. DTH-induced mice were treated with THC or vehicle (n = 5 per experimental group) and the PopLN assessed for the early activation marker CD69. (a & b) Flow cytometry dot plots (gated on live cells) showing (a) CD3 + CD69+ and (b) CD19 + CD69+ cells. Cells from the draining LN were cultured in the presence of PMA, Inomycin, and Golgi Plug (4 hours) and stained for IL-17a secreting cells. (c) Flow cytometry dot plots (gated on CD4) showing CD4 + IL-17a+. Representative data from replicate experiments. (PDF 292 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Jessica M. Sido
    • 1
  • Austin R. Jackson
    • 1
  • Prakash S. Nagarkatti
    • 1
  • Mitzi Nagarkatti
    • 1
    • 2
    Email author
  1. 1.Department of Pathology, Microbiology, and ImmunologyUniversity of South Carolina School of MedicineColumbiaUSA
  2. 2.WJB Dorn Veterans Affairs Medical CenterColumbiaUSA

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