title: Calreticulin is an interleukin-3-sensitive calcium-binding protein in human basophil leukocytes.
abstract: BACKGROUND
  IL-3 enhances basophil histamine release upon stimulation with any known secretagogue. The molecular mechanism behind this regulation is not known, although some observations suggest that IL-3 modulates the calcium part of the signal transduction mechanism. The inhibitory action of glucocorticoids on basophils can be reversed by stimulation with IL-3.
  
  
  METHODS
  Calcium-binding proteins in the basophil cell line KU812 were identified by two-dimensional gel electrophoresis, Calcium-overlay assay, N-terminal sequence analysis, and mass spectometry. The presence of the same proteins in purified human basophil leukocytes was established by comigration of KU812 and human basophil proteins on the two-dimensional gels. The expression of the calcium-binding proteins in the absence and presence of IL-3 and/or anti-IgE was determined by densitometric measurement of the spots on the two-dimensional gels.
  
  
  RESULTS
  Calreticulin was identified on the two-dimensional gel of KU812 proteins. A protein with exactly the same migration pattern was found on the gels of proteins from purified human basophils. Immunoblotting with a specific antihuman calreticulin antibody confirmed that this protein was calreticulin. Subsequent analysis showed that the expression of calreticulin in the basophils is upregulated twofold upon stimulation with rhIL-3, even in doses below those needed for enhancement of histamine release.
  
  
  CONCLUSIONS
  The expression of calreticulin in human basophil leukocytes is regulated by IL-3. Calreticulin is known to modulate IP3-dependent Ca2+ influx in different cell systems, and calreticulin overexpression inhibits steroid-induced transcriptional activation. Therefore, modulation of calreticulin expression may be one mechanism by which IL-3 exerts its effects on human basophils.
authors.affiliation: Department of Pediatrics, Aarhus University Hospital, Denmark.
authors.forename: J M
authors.initials: JM
authors.lastname: Lyngholm
authors.forename: H V
authors.initials: HV
authors.lastname: Nielsen
authors.forename: M
authors.initials: M
authors.lastname: Holm
authors.forename: P O
authors.initials: PO
authors.lastname: Schiøtz
authors.forename: A H
authors.initials: AH
authors.lastname: Johnsen
mesh_terms: D001491:Basophils / Q000187:drug effects / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D037282:Calreticulin; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D015180:Electrophoresis, Gel, Two-Dimensional; D015870:Gene Expression; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology*; D012261:Ribonucleoproteins / Q000378:metabolism*
chemical_list: D002135:Calcium-Binding Proteins; D037282:Calreticulin; D007377:Interleukin-3; D012261:Ribonucleoproteins; D002118:Calcium