title: Clinical Experience With Modified, Single-Tube T-Cell Receptor Vβ Flow Cytometry Analysis for T-Cell Clonality.
abstract: OBJECTIVE
  Analysis of T-cell clonality using the IO Test Beta Mark (Beckman Coulter, Fullerton, CA) flow cytometry assay is cumbersome owing to the required eight-part analyses that limit its clinical utility. Here, we describe a simple modification that may improve its clinical utility.
  
  
  METHODS
  By simply combining all T-cell receptor (TCR) Vβ antibodies into one reagent tube, we show that T-cell repertoire analysis can be efficiently performed with an increased potential to closely link the aberrant immunophenotype of a neoplastic T-cell clone to its apparent clonal TCR Vβ restriction. We review our clinical experience and include flow cytometry sorting studies in conjunction with TRG gene rearrangement analysis to demonstrate the feasibility of this approach.
  
  
  RESULTS
  We show in representative cases that this modified approach permits evaluation of TCR Vβ clonality in T-cell neoplasms without a priori knowledge of the TCR Vβ isoform restriction and that this approach is suitable for samples containing a minor population of neoplastic T cells, hypocellular samples such as skin biopsy specimens and body fluids, and fine-needle aspirations. We estimate a false-positive rate for identifying a clonal population for this approach to be less than 6.2%.
  
  
  CONCLUSIONS
  Our modified approach of TCR Vβ analysis could improve routine clinical evaluation for abnormal T-cell populations by flow cytometry.
authors.affiliation: From the Department of Laboratory Medicine, University of Washington, Seattle;
authors.forename: David
authors.initials: D
authors.lastname: Wu
authors.affiliation: From the Department of Laboratory Medicine, University of Washington, Seattle;
authors.forename: Melanie M
authors.initials: MM
authors.lastname: Anderson
authors.affiliation: Biostatistics and Biomathematics, Fred Hutchinson Cancer Research Center, Public Health Sciences, Seattle, WA.
authors.forename: Megan
authors.initials: M
authors.lastname: Othus
authors.affiliation: From the Department of Laboratory Medicine, University of Washington, Seattle; woodbl@uw.edu.
authors.forename: Brent L
authors.initials: BL
authors.lastname: Wood
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002999:Clone Cells / Q000166:cytology; D005189:False Positive Reactions; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor / Q000235:genetics*; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D008232:Lymphoproliferative Disorders / Q000175:diagnosis; D008297:Male; D008875:Middle Aged; D016693:Receptors, Antigen, T-Cell, alpha-beta
chemical_list: D016693:Receptors, Antigen, T-Cell, alpha-beta
keywords: Flow cytometry; IO Test Beta Mark kit; T-cell clonality; T-cell receptor analysis; TCR Vβ