Impaired NFAT and NFκB Activation Are Involved in Suppression of CD40 Ligand Expression by Δ(9)-tetrahydrocannabinol in Human CD4(+) T Cells

Toxicol Appl Pharmacol. 2013 Nov 15;273(1):209-18. doi: 10.1016/j.taap.2013.08.023. Epub 2013 Aug 30.

Abstract

We have previously reported that Δ(9)-tetrahydrocannabinol (Δ(9)-THC), the main psychoactive cannabinoid in marijuana, suppresses CD40 ligand (CD40L) expression by activated mouse CD4(+) T cells. CD40L is involved in pathogenesis of many autoimmune and inflammatory diseases. In the present study, we investigated the molecular mechanism of Δ(9)-THC-mediated suppression of CD40L expression using peripheral blood human T cells. Pretreatment with Δ(9)-THC attenuated CD40L expression in human CD4(+) T cells activated by anti-CD3/CD28 at both the protein and mRNA level, as determined by flow cytometry and quantitative real-time PCR, respectively. Electrophoretic mobility shift assays revealed that Δ(9)-THC suppressed the DNA-binding activity of both NFAT and NFκB to their respective response elements within the CD40L promoter. An assessment of the effect of Δ(9)-THC on proximal T cell-receptor (TCR) signaling induced by anti-CD3/CD28 showed significant impairment in the rise of intracellular calcium, but no significant effect on the phosphorylation of ZAP70, PLCγ1/2, Akt, and GSK3β. Collectively, these findings identify perturbation of the calcium-NFAT and NFκB signaling cascade as a key mechanistic event by which Δ(9)-THC suppresses human T cell function.

Keywords: CD40L; Intracellular calcium; NFAT; NFκB; T cell-receptor signaling; Δ(9)-tetrahydrocannabinol (Δ(9)-THC).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CD28 Antigens / metabolism
  • CD4-Positive T-Lymphocytes / cytology
  • CD4-Positive T-Lymphocytes / drug effects*
  • CD4-Positive T-Lymphocytes / metabolism
  • CD40 Ligand / antagonists & inhibitors
  • CD40 Ligand / genetics*
  • CD40 Ligand / metabolism
  • Calcium / analysis
  • Cannabinoids / chemistry
  • Dronabinol / pharmacology*
  • Female
  • Flow Cytometry
  • Gene Expression Regulation
  • Glycogen Synthase Kinase 3 / genetics
  • Glycogen Synthase Kinase 3 / metabolism
  • Glycogen Synthase Kinase 3 beta
  • Humans
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / drug effects
  • Mice
  • Mice, Inbred C57BL
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / genetics*
  • NF-kappa B / metabolism
  • NFATC Transcription Factors / antagonists & inhibitors
  • NFATC Transcription Factors / genetics*
  • NFATC Transcription Factors / metabolism
  • Phospholipase C gamma / genetics
  • Phospholipase C gamma / metabolism
  • Phosphorylation
  • Proto-Oncogene Proteins c-akt / genetics
  • Proto-Oncogene Proteins c-akt / metabolism
  • Real-Time Polymerase Chain Reaction
  • Signal Transduction
  • ZAP-70 Protein-Tyrosine Kinase / genetics
  • ZAP-70 Protein-Tyrosine Kinase / metabolism

Substances

  • CD28 Antigens
  • Cannabinoids
  • NF-kappa B
  • NFATC Transcription Factors
  • CD40 Ligand
  • Dronabinol
  • ZAP-70 Protein-Tyrosine Kinase
  • ZAP70 protein, human
  • GSK3B protein, human
  • Glycogen Synthase Kinase 3 beta
  • Gsk3b protein, mouse
  • Proto-Oncogene Proteins c-akt
  • Glycogen Synthase Kinase 3
  • Phospholipase C gamma
  • Calcium