title: Blockade of nicotinic and muscarinic receptors facilitates spontaneous migration of human peripheral granulocytes: failure in cystic fibrosis.
abstract: OBJECTIVE
  Circulating leucocytes express muscarinic (m) and nicotinic (n) receptors and synthesize acetylcholine (ACh) regulating various cell functions. Leucocytes from patients with cystic fibrosis contain less ACh; therefore it was tested whether the regulation of cellular functions like migration differed from healthy volunteers.
  
  
  METHODS
  Peripheral blood (10-20 ml) was used, leucocytes were isolated by Ficoll® gradient and the commercial MIGRATEST® combined with flow cytometric analysis was applied (pore size 3 μm).
  
  
  RESULTS
  In the absence of test substances 4900±1800 (n=10) leucocytes migrated within a time period of 2 h. In the presence of tubocurarine (TC, 30 μM) the cell number increased to 7500±2700 [n=10] corresponding to an increase of 162±20% (mean of individual experiments; p<0.02). Atropine (1 μM) was not effective (120±17%, n=7). Simultaneous application of atropine and TC produced a slightly higher effect than TC alone (185±23%; n=8); a 10-fold increase of TC and atropine resulted to a somewhat stronger effect (248±39%; n=8). When migration time was reduced to 30 min or the chemoattractant fMLP (0.05 μM) present neither atropine nor TC affected migration. Granulocytes isolated from patients with cystic fibrosis did not respond (2h migration) to 30 μM TC (control: 5180±1400 cells [n=10]; TC: 5800±1400 [n=10]). Also in the presence of atropine (1 μM) and TC (30 μM) a significant effect was not detected (5800±1300 [n=10]).
  
  
  CONCLUSIONS
  Auto-paracrine acetylcholine limits the migration of unstimulated peripheral granulocytes. This effect is impaired in cystic fibrosis most likely because of a reduced endogenous cholinergic tone.
authors.affiliation: Institute of Pathology, Johannes Gutenberg University Mainz, D-55101 Mainz, Germany. wessler@uni-mainz.de
authors.forename: Ignaz
authors.initials: I
authors.lastname: Wessler
authors.forename: Stefanie
authors.initials: S
authors.lastname: Neumann
authors.forename: Michael
authors.initials: M
authors.lastname: Razen
authors.forename: Fred
authors.initials: F
authors.lastname: Zepp
authors.forename: Charles James
authors.initials: CJ
authors.lastname: Kirkpatrick
mesh_terms: D000293:Adolescent; D000328:Adult; D054441:Cell Migration Assays, Leukocyte; D002465:Cell Movement / Q000187:drug effects; D002648:Child; D018680:Cholinergic Antagonists / Q000494:pharmacology*; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000473:pathology*; D005260:Female; D006098:Granulocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D008297:Male; D011976:Receptors, Muscarinic / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000378:metabolism*; D055815:Young Adult
chemical_list: D018680:Cholinergic Antagonists; D011976:Receptors, Muscarinic; D011978:Receptors, Nicotinic