title: Pimecrolimus reduces eosinophil activation associated with calcium mobilization.
abstract: BACKGROUND
  Pimecrolimus is a calcineurin inhibitor that inhibits T cell and mast cell activation and effectively treats atopic dermatitis. However, its effects on eosinophils, a cell type implicated in allergic disease pathology, are unknown. Therefore, we examined the effects of pimecrolimus on eosinophil superoxide anion production, degranulation and survival.
  
  
  METHODS
  Purified eosinophils from normal or atopic donors were incubated with serial dilutions of pimecrolimus (microM to nM) and then stimulated with platelet activating factor (PAF), interleukin 5 (IL5), secretory immunoglobulin A (sIgA) or Alternaria alternata (Alt) fungus extract. Eosinophil activation was monitored by cytochrome c reduction resulting from superoxide anion production and by a 2-site immunoassay for eosinophil-derived neurotoxin (EDN) in cellular supernatants, as a marker of degranulation. Eosinophil survival was measured by propidium iodide exclusion using flow cytometry after 4 days in culture.
  
  
  RESULTS
  Normal and atopic eosinophil superoxide anion production induced by PAF, and associated with increased intracellular calcium, was inhibited up to 37% with 1 microM pimecrolimus. However, superoxide anion production induced by IL5 and sIgA was not consistently inhibited. EDN release, which ultimately depends on calcium, was inhibited about 30% with PAF, IL5 and sIgA stimulation for normal and atopic donor eosinophils. Furthermore, calcium-dependent Alt-induced EDN release was inhibited up to 49% with nanomolar pimecrolimus. Finally, increased eosinophil survival promoted by IL5 and sIgA was not influenced by pimecrolimus.
  
  
  CONCLUSIONS
  Pimecrolimus moderately inhibits eosinophil superoxide anion production and EDN release associated with calcium mobilization, which may contribute to its efficacy in treating atopic dermatitis.
authors.affiliation: Allergic Diseases Research Laboratory, Department of Dermatology, Mayo Clinic, Guggenheim 4-94, 200 First Street Southwest, Rochester, MN 55905, USA. plager.douglas@mayo.edu
authors.forename: Douglas A
authors.initials: DA
authors.lastname: Plager
authors.forename: Susan A
authors.initials: SA
authors.lastname: Henke
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Matsuwaki
authors.forename: Arvind
authors.initials: A
authors.lastname: Madaan
authors.forename: Diane L
authors.initials: DL
authors.lastname: Squillace
authors.forename: Ross A
authors.initials: RA
authors.lastname: Dierkhising
authors.forename: Hirohito
authors.initials: H
authors.lastname: Kita
mesh_terms: D065095:Calcineurin Inhibitors; D002118:Calcium / Q000276:immunology / Q000378:metabolism; D015550:Cell Degranulation / Q000187:drug effects* / Q000276:immunology; D002470:Cell Survival / Q000187:drug effects / Q000276:immunology; D002478:Cells, Cultured; D047208:Eosinophil-Derived Neurotoxin / Q000032:analysis / Q000276:immunology*; D004804:Eosinophils / Q000187:drug effects* / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007071:Immunoglobulin A, Secretory / Q000494:pharmacology; D015848:Interleukin-5 / Q000494:pharmacology; D010972:Platelet Activating Factor / Q000494:pharmacology; D013481:Superoxides / Q000032:analysis / Q000276:immunology; D016559:Tacrolimus / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: D065095:Calcineurin Inhibitors; D007071:Immunoglobulin A, Secretory; D015848:Interleukin-5; D010972:Platelet Activating Factor; D013481:Superoxides; C117268:pimecrolimus; D047208:Eosinophil-Derived Neurotoxin; D002118:Calcium; D016559:Tacrolimus