title: Effect of N-methyl-D-aspartate receptor antagonist on T helper cell differentiation induced by phorbol-myristate-acetate and ionomycin.
abstract: OBJECTIVE
  To investigate the effect of N-methyl-D-aspartate (NMDA) receptor antagonist on T helper (TH) cell differentiation and intracellular transcriptional factors in vitro.
  
  
  METHODS
  Ten male healthy volunteers (aged 20-45 y, BMI 18-25) were enrolled in this study. Twenty milliliters peripheral blood was collected in the morning from fasted volunteers and peripheral blood mononuclear cells (PBMC) were isolated. PBMC were incubated with phorbol-myristate-acetate (25 ng/ml) (PMA) plus ionomycin (1 μg/ml) in the presence of ketamine or MK-801 at 37 °C. TH subsets, supernatant interferon γ (IFN-γ), interleukin 4 (IL-4), and intracellular transcriptional factors T-bet and GATA3 were analyzed 4 h later.
  
  
  RESULTS
  The number of TH0 cells was kept constant and at baseline before PMA and ionomycin stimulation in each group. TH1 cells, TH2 cells, IFN-γ and IL-4 levels were significantly increased after PMA and ionomycin stimulation. Ketamine and MK-801 decreased TH1 cells, TH2 cells, IFN-γ and IL-4 levels but increased the ratio of TH1/TH2 and IFN-γ/IL-4 in the presence of PMA and ionomycin. Ketamine or MK-801 alone had no effect on either of them. T-bet and GATA3 activities in PBMC were significantly increased after PMA and ionomycin stimulation. Ketamine and MK-801 decreased T-bet and GATA3 activities but increased the ratio of T-bet/GATA3 following PMA and ionomycin stimulation. Ketamine or MK-801 alone had no effect on the activity of T-bet, GATA3 or T-bet/GATA3.
  
  
  CONCLUSIONS
  NMDA antagonist can suppress TH cell differentiation and subsequent cytokines production but increase TH1/TH2 ratio following PMA and ionomycin stimulation, it may be related to its regulation on T-bet and GATA3 activities.
authors.affiliation: Department of Anesthesiology, The First Affiliated Hospital with Nanjing Medical University, No. 300, Guangzhou road, Nanjing 210029, PR China.
authors.forename: Mei
authors.initials: M
authors.lastname: Gao
authors.forename: Wenjie
authors.initials: W
authors.lastname: Jin
authors.forename: Yanning
authors.initials: Y
authors.lastname: Qian
authors.forename: Lulu
authors.initials: L
authors.lastname: Ji
authors.forename: Genbao
authors.initials: G
authors.lastname: Feng
authors.forename: Jie
authors.initials: J
authors.lastname: Sun
mesh_terms: D000328:Adult; D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects*; D016291:Dizocilpine Maleate / Q000494:pharmacology; D024202:Electrophoretic Mobility Shift Assay; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology*; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D015759:Ionomycin / Q000494:pharmacology*; D007649:Ketamine / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D015345:Oligonucleotide Probes; D016194:Receptors, N-Methyl-D-Aspartate / Q000037:antagonists & inhibitors*; D006377:T-Lymphocytes, Helper-Inducer / Q000166:cytology / Q000187:drug effects*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
chemical_list: D018691:Excitatory Amino Acid Antagonists; D015345:Oligonucleotide Probes; D016194:Receptors, N-Methyl-D-Aspartate; D015847:Interleukin-4; D015759:Ionomycin; D007649:Ketamine; D016291:Dizocilpine Maleate; D007371:Interferon-gamma; D013755:Tetradecanoylphorbol Acetate