title: Basophil markers for identification and activation in the indirect basophil activation test by flow cytometry for diagnosis of autoimmune urticaria.
abstract: The indirect basophil activation test using flow cytometry is a promising tool for autoimmune urticaria diagnosis. We aimed to identify better donor basophils (from atopic vs. non-atopic donors and interleukin-3 primed vs. unprimed basophils) and improve basophil identification and activation markers (eotaxin CC chemokine receptor-3 [CCR3] vs. CD123 and CD63 vs. CD203c).
  
  
  
  Donor basophils were obtained from non-atopic and atopic group O donors. Positive control sera were artificially prepared to simulate autoimmune urticaria patients' sera. Patient sera were obtained from nine children with chronic urticaria. Assay sensitivity was compared among each variation by using positive control sera (n=21), applying cutoff values defined from negative control sera (n=20).
  
  
  
  For basophil identification, a combination of CCR3 and CD123 markers revealed a higher correlation with automated complete blood count (r=0.530) compared with that observed using CD123 (r=0.498) or CCR3 alone (r=0.195). Three activation markers on the atopic donor basophils attained 100% assay sensitivity: CD203c on unprimed basophils, CD63+CD203+ or CD63 alone on primed basophils; however, these markers on the non-atopic donor basophils attained lower assay sensitivity.
  
  
  
  For basophil identification markers, a combination of CD123 and CCR3 is recommended, while CD123 alone may be used as an alternative. Donor basophils should be obtained from an atopic donor. For basophil activation markers, either CD203c alone on unprimed basophils or CD203c and CD63 on primed basophils are recommended, while CD63 alone on primed basophils may be used as an alternative.
authors.affiliation: Department of Clinical Pathology, Kyungpook National University School of Medicine, Daegu, Korea.
authors.forename: Zehwan
authors.initials: Z
authors.lastname: Kim
authors.affiliation: Department of Pediatrics, Kyungpook National University School of Medicine, Daegu, Korea.
authors.forename: Bong Seok
authors.initials: BS
authors.lastname: Choi
authors.affiliation: Department of Emergency Medicine, Kyungpook National University School of Medicine, Daegu, Korea.
authors.forename: Jong Kun
authors.initials: JK
authors.lastname: Kim
authors.affiliation: Department of Clinical Pathology, Kyungpook National University School of Medicine, Daegu, Korea. wondi@knu.ac.kr.
authors.forename: Dong Il
authors.initials: DI
authors.lastname: Won
mesh_terms: D001327:Autoimmune Diseases / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D001491:Basophils / Q000276:immunology* / Q000378:metabolism; D015415:Biomarkers / Q000097:blood; D002648:Child; D005434:Flow Cytometry; D006801:Humans; D053650:Interleukin-3 Receptor alpha Subunit / Q000097:blood; D008297:Male; D054397:Receptors, CCR3 / Q000097:blood; D014581:Urticaria / Q000097:blood / Q000175:diagnosis* / Q000276:immunology
chemical_list: D015415:Biomarkers; C515350:CCR3 protein, human; C115022:IL3RA protein, human; D053650:Interleukin-3 Receptor alpha Subunit; D054397:Receptors, CCR3
keywords: Autoimmune urticaria; Basophil; Chronic urticaria; Donor; Flow cytometry; Markers