title: Flow cytometric immunophenotypic assessment of T-cell clonality by vβ repertoire analysis in fine-needle aspirates and cerebrospinal fluid.
abstract: Flow cytometric T-cell receptor V(β) repertoire analysis (TCR-V(β)-R) is a sensitive method to detect T-cell clonality; however, its implementation in low-cellularity specimens has not been established. We developed a strategy to use TCR-V(β)-R in cerebrospinal fluid (CSF) and fine-needle aspirate (FNA) specimens. Initially, full TCR-V(β)-R was evaluated in diagnostic/screening specimens from 8 patients with T-cell neoplasia to determine tumor-specific TCR-V(β) protein expression. Subsequently, an abbreviated, patient-specific TCR-V(β)-R evaluation was performed in 17 paucicellular specimens from the patients (8 CSF, 9 FNA) for staging and monitoring of minimal residual disease (MRD). A single cocktail containing 3 anti-V(β) antibodies (1 tumor-specific and 2 negative controls) in combination with other antibodies chosen to help gate on atypical T cells is highly sensitive and specific for detecting low-level neoplastic T-cell involvement in paucicellular specimens. This TCR-V(β)-R strategy is valuable in staging and evaluating MRD in patients with T-cell non-Hodgkin lymphoma.
authors.affiliation: Flow Cytometry Unit, Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Prashant
authors.initials: P
authors.lastname: Tembhare
authors.forename: Constance M
authors.initials: CM
authors.lastname: Yuan
authors.forename: John C
authors.initials: JC
authors.lastname: Morris
authors.forename: John E
authors.initials: JE
authors.lastname: Janik
authors.forename: Armando C
authors.initials: AC
authors.lastname: Filie
authors.forename: Maryalice
authors.initials: M
authors.lastname: Stetler-Stevenson
mesh_terms: D044963:Biopsy, Fine-Needle; D002471:Cell Transformation, Neoplastic; D002555:Cerebrospinal Fluid / Q000276:immunology*; D002999:Clone Cells / Q000276:immunology; D004273:DNA, Neoplasm / Q000032:analysis; D005434:Flow Cytometry / Q000379:methods*; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor / Q000235:genetics / Q000276:immunology*; D019674:Genes, T-Cell Receptor beta / Q000235:genetics / Q000276:immunology*; D006801:Humans; D016130:Immunophenotyping; D016399:Lymphoma, T-Cell / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000235:genetics / Q000276:immunology; D018365:Neoplasm, Residual / Q000134:cerebrospinal fluid / Q000175:diagnosis / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000032:analysis; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D004273:DNA, Neoplasm; D016693:Receptors, Antigen, T-Cell, alpha-beta