Anti-TRBC1 Antibody-Based Flow Cytometric Detection of T-Cell Clonality: Standardization of Sample Preparation and Diagnostic Implementation

Noemí Muñoz-García; Margarida Lima; Neus Villamor; F. Javier Morán-Plata; Susana Barrena; Sheila Mateos; Carolina Caldas; Ana Balanzategui; Miguel Alcoceba; Alejandro Domínguez; Fabio Gómez; Anton W. Langerak; Jacques J. M. van Dongen; Alberto Orfao; Julia Almeida

doi:10.3390/cancers13174379

Cancers (Aug 2021)

Anti-TRBC1 Antibody-Based Flow Cytometric Detection of T-Cell Clonality: Standardization of Sample Preparation and Diagnostic Implementation

Noemí Muñoz-García,
Margarida Lima,
Neus Villamor,
F. Javier Morán-Plata,
Susana Barrena,
Sheila Mateos,
Carolina Caldas,
Ana Balanzategui,
Miguel Alcoceba,
Alejandro Domínguez,
Fabio Gómez,
Anton W. Langerak,
Jacques J. M. van Dongen,
Alberto Orfao,
Julia Almeida

Affiliations

Noemí Muñoz-García: Translational and Clinical Research Program, Centro de Investigación del Cáncer and IBMCC (CSIC-University of Salamanca), Cytometry Service, NUCLEUS, Department of Medicine, University of Salamanca (USAL) and Institute of Biomedical Research of Salamanca (IBSAL), 37007 Salamanca, Spain
Margarida Lima: Department of Hematology, Laboratory of Cytometry, Hospital de Santo António, Centro Hospitalar do Porto, 4099-001 Porto, Portugal
Neus Villamor: Biomedical Research Networking Centre Consortium of Oncology (CIBERONC), Instituto de Salud Carlos III, 28029 Madrid, Spain
F. Javier Morán-Plata: Translational and Clinical Research Program, Centro de Investigación del Cáncer and IBMCC (CSIC-University of Salamanca), Cytometry Service, NUCLEUS, Department of Medicine, University of Salamanca (USAL) and Institute of Biomedical Research of Salamanca (IBSAL), 37007 Salamanca, Spain
Susana Barrena: Translational and Clinical Research Program, Centro de Investigación del Cáncer and IBMCC (CSIC-University of Salamanca), Cytometry Service, NUCLEUS, Department of Medicine, University of Salamanca (USAL) and Institute of Biomedical Research of Salamanca (IBSAL), 37007 Salamanca, Spain
Sheila Mateos: Translational and Clinical Research Program, Centro de Investigación del Cáncer and IBMCC (CSIC-University of Salamanca), Cytometry Service, NUCLEUS, Department of Medicine, University of Salamanca (USAL) and Institute of Biomedical Research of Salamanca (IBSAL), 37007 Salamanca, Spain
Carolina Caldas: Translational and Clinical Research Program, Centro de Investigación del Cáncer and IBMCC (CSIC-University of Salamanca), Cytometry Service, NUCLEUS, Department of Medicine, University of Salamanca (USAL) and Institute of Biomedical Research of Salamanca (IBSAL), 37007 Salamanca, Spain
Ana Balanzategui: Biomedical Research Networking Centre Consortium of Oncology (CIBERONC), Instituto de Salud Carlos III, 28029 Madrid, Spain
Miguel Alcoceba: Biomedical Research Networking Centre Consortium of Oncology (CIBERONC), Instituto de Salud Carlos III, 28029 Madrid, Spain
Alejandro Domínguez: Centro de Salud Miguel Armijo, Sanidad de Castilla y León (SACYL), 37007 Salamanca, Spain
Fabio Gómez: Centro de Salud Miguel Armijo, Sanidad de Castilla y León (SACYL), 37007 Salamanca, Spain
Anton W. Langerak: Department of Immunology, Laboratory Medical immunology, Erasmus MC, University Medical Center Rotterdam, 3015 GD Rotterdam, The Netherlands
Jacques J. M. van Dongen: Department of Immunology, Leiden University Medical Center (LUMC), 2333 ZA Leiden, The Netherlands
Alberto Orfao: Translational and Clinical Research Program, Centro de Investigación del Cáncer and IBMCC (CSIC-University of Salamanca), Cytometry Service, NUCLEUS, Department of Medicine, University of Salamanca (USAL) and Institute of Biomedical Research of Salamanca (IBSAL), 37007 Salamanca, Spain
Julia Almeida: Translational and Clinical Research Program, Centro de Investigación del Cáncer and IBMCC (CSIC-University of Salamanca), Cytometry Service, NUCLEUS, Department of Medicine, University of Salamanca (USAL) and Institute of Biomedical Research of Salamanca (IBSAL), 37007 Salamanca, Spain

DOI: https://doi.org/10.3390/cancers13174379
Journal volume & issue: Vol. 13, no. 17
p. 4379

Abstract

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A single antibody (anti-TRBC1; JOVI-1 antibody clone) against one of the two mutually exclusive T-cell receptor β-chain constant domains was identified as a potentially useful flow-cytometry (FCM) marker to assess Tαβ-cell clonality. We optimized the TRBC1-FCM approach for detecting clonal Tαβ-cells and validated the method in 211 normal, reactive and pathological samples. TRBC1 labeling significantly improved in the presence of CD3. Purified TRBC1+ and TRBC1− monoclonal and polyclonal Tαβ-cells rearranged TRBJ1 in 44/47 (94%) and TRBJ1+TRBJ2 in 48 of 48 (100%) populations, respectively, which confirmed the high specificity of this assay. Additionally, TRBC1+/TRBC1− ratios within different Tαβ-cell subsets are provided as reference for polyclonal cells, among which a bimodal pattern of TRBC1-expression profile was found for all TCRVβ families, whereas highly-variable TRBC1+/TRBC1− ratios were observed in more mature vs. naïve Tαβ-cell subsets (vs. total T-cells). In 112/117 (96%) samples containing clonal Tαβ-cells in which the approach was validated, monotypic expression of TRBC1 was confirmed. Dilutional experiments showed a level of detection for detecting clonal Tαβ-cells of ≤10−4 in seven out of eight pathological samples. These results support implementation of the optimized TRBC1-FCM approach as a fast, specific and accurate method for assessing T-cell clonality in diagnostic-FCM panels, and for minimal (residual) disease detection in mature Tαβ+ leukemia/lymphoma patients.

Published in Cancers

ISSN: 2072-6694 (Online)
Publisher: MDPI AG
Country of publisher: Switzerland
LCC subjects: Medicine: Internal medicine: Neoplasms. Tumors. Oncology. Including cancer and carcinogens
Website: https://www.mdpi.com/journal/cancers/

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