title: Modulation of immune functions in polymorphonuclear neutrophils induced by physostigmine, but not neostigmine, independent of cholinergic neurons.
abstract: BACKGROUND
  Cholinesterase inhibitors (Ch-I) improve survival in experimental sepsis consistent with activation of the cholinergic-anti-inflammatory-pathway. So far, less is known about whether Ch-I have a direct immunomodulatory effect on immune cells (polymorphonuclear neutrophils, PMN) in the absence of cholinergic neurons. We investigated the concentration-response-effects of physostigmine and neostigmine on the oxidative burst activity (human and rat PMN) and the expression of adhesion molecules on the surface of human PMN under in vitro conditions.
  
  
  METHODS
  PMN from 10 healthy humans or 10 rats were pretreated with 2, 10, 24, 97 μM physostigmine or 3, 15, 30, 150 μM neostigmine, primed with tumor-necrosis-factor-alpha (TNF-alpha) followed by stimulation with n-formyl-methionyl-leucylphenylalanine (fMLP) or stimulated with phorbol-12-myristate-13-acetate (PMA). Human and rat samples were assessed by flow cytometry for the generation of oxidative free radicals. Stimulated human PMN were additionally incubated with antibodies against Mac-1 (CD11b) or L-selectin (CD62l).
  
  
  RESULTS
  Physostigmine and neostigmine did not alter oxidative burst activity or the expression of adhesion molecules of PMN induced by receptor-dependent activators like fMLP or TNF-alpha/fMLP (rat and human PMN, p=n.s.). Physostigmine, but not neostigmine, inhibited the protein-kinase-C-mediated oxidative burst activity by PMA in a dose-dependent manner (rat and human PMN, p<0.05). Physostigmine, in the concentration range tested, suppressed the expression of CD11b following stimulation with PMA not significantly (human PMN: control: 63.1±10.7 vs. 97 μM physostigmine: 49.9±12.8 MESF, p=n.s.).
  
  
  CONCLUSIONS
  While neostigmine has no effect on functional and phenotypic changes of PMN, the lipid soluble Ch-I physostigmine causes a dose dependent reduction in PMA-induced oxidative burst, independent of neuronal released acetylcholine.
authors: Department of Anaesthesiology, University of Regensburg, Regensburg, Germany. diane.bitzinger@ukr.de
authors: Diane I
authors: DI
authors: Bitzinger
authors: York A
authors: YA
authors: Zausig
authors: Christoph
authors: C
authors: Paech
authors: Michael
authors: M
authors: Gruber
authors: Hendrik
authors: H
authors: Busse
authors: Barbara
authors: B
authors: Sinner
authors: Bernhard M
authors: BM
authors: Graf
authors: Benedikt
authors: B
authors: Trabold
mesh_terms: D000818:Animals; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000187:drug effects; D002478:Cells, Cultured; D059329:Cholinergic Neurons; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D006801:Humans; D019041:L-Selectin / Q000276:immunology; D016177:Macrophage-1 Antigen / Q000276:immunology; D008297:Male; D009240:N-Formylmethionine Leucyl-Phenylalanine; D009388:Neostigmine / Q000494:pharmacology*; D009504:Neutrophils / Q000187:drug effects*; D010830:Physostigmine / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D016897:Respiratory Burst / Q000187:drug effects*; D013755:Tetradecanoylphorbol Acetate; D014409:Tumor Necrosis Factor-alpha
chemical_list: D015815:Cell Adhesion Molecules; D002800:Cholinesterase Inhibitors; D016177:Macrophage-1 Antigen; D014409:Tumor Necrosis Factor-alpha; D019041:L-Selectin; D009388:Neostigmine; D009240:N-Formylmethionine Leucyl-Phenylalanine; D010830:Physostigmine; D013755:Tetradecanoylphorbol Acetate