TCRbeta1/TCRbeta2 (TRBC1/TRBC2) antibody pair for determining T-cell monotypia as a surrogate for clonality in formalin-fixed paraffin-embedded material

Sep 23, 2026

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Aim: T-cell lymphomas are often histologically indistinguishable from benign T-cell infiltrates, making clonality testing important yet slow and costly. Existing workflows rely on complex multiplexed PCR with limited spatial context and specialist interpretation. Methods: The study generated highly specific monoclonal antibodies targeting TCRβ1 and TCRβ2, encoded by TRBC1 and TRBC2, and evaluated immunohistochemical detection in formalin-fixed, paraffin-embedded tissue. Results: Single staining was assessed across lymphoma and benign T-cell samples, with double staining examples. Automated cell counting enabled accurate TCRβ2:TCRβ1 ratio calculation. Conclusion: The assay offers a timely, automated approach analogous to kappa/lambda testing.

LanguageEnglish

CategoryHealthcare

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Pages8

Reading Time20 min

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Abstract

Introduction

What is already known on this topic

What this study adds

How this study might affect research, practice or policy

Original research TCRbeta1/TCRbeta2 (TRBC1/TRBC2) antibody pair for determining T-cell monotypia as a surrogate forclonality in formalin-fixed paraffin-embedded material Anuradha Kaistha, 1 Jinlong J Situ, 1 Shelley Evans, 1 Margaret Ashton-Key,2 Graham Ogg,3 Elizabeth Soilleux 1 1Department of Pathology, University of Cambridge, Cambridge, UK 2Department of Cellular Pathology, Southampton University Hospitals NHS Trust, Southampton, UK 3MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, UK Correspondence to Professor Elizabeth Soilleux; [ejs17@cam.ac.uk](ejs17@cam.ac.uk) AK and JJS contributed equally. Received 13 February 2026 Accepted 12 April 2026 © Author(s) (or their employer(s)) 2026. Re-use permitted under CC BY. Published by BMJ Group. To cite: Kaistha A, Situ JJ, Evans S, et al. J Clin Pathol Epub ahead of print: [please include Day Month Year] . doi:10 . 1136/jcp-2026- 210684 ABSTRACT Aim T-cell lymphomas are often histologically indistinguishable from benign T-cell infiltrates. Clonality testing is frequently required for diagnosis. It lacks the spatial context and is slow and expensive, relying on complex multiplexed PCR reactions, interpreted by scientists or pathologists with specialist molecular training. We set out to make monoclonal antibodies to develop a novel immunohistochemical test for T-cell lymphoma, analogous to the kappa/lambda assay for B-cell and plasma cell neoplasms. Methods We developed a pair of highly specific monoclonal antibodies against the two alternatively used but very similar T-cell receptor β constant regions, TCRβ1 and TCRβ2 (encoded by the TRBC1 and TRBC2 gene segments). We demonstrate the feasibility of immunohistochemical detection of TCRβ1 and TCRβ2 in formalin-fixed, paraffin-embedded tissue as a novel diagnostic strategy for T-cell lymphomas. Results Single immunostaining results are presented for 13 T-cell lymphomas and 8 benign T-cell populations, together with illustrative examples of TCRβ1/2 double immunostaining. Finally, we show that this single immunostaining is amenable to automated cell counting, permitting accurate calculation of the TCRβ2:TCRβ1 ratio. Conclusion This novel assay can be used in a similar way to the kappa/lambda assay for B-cell and plasma cell neoplasms. INTRODUCTION T-cell lymphomas account for 10%–15% of lymphomas and frequently have a rapid and aggressive course.1 They are difficult, time-consuming and expensive to diagnose, so patients often require multiple biopsies, with months or years-long diagnostic delays impacting survival (reviewed in Soilleux et al).1 T-cell lymphomas composed of small-to-mediumsized cells (eg, peripheral T-cell lymphoma, angioimmunoblastic lymphoma and mycosis fungoides)2 are particularly difficult to distinguish, both morphologically and immunophenotypically, from benign lymphocytic infiltrates. Diagnosis often necessitates multiplexed PCR T-cell clonality analysis of DNA extracted from tissue, a complex test interpreted by specially trained staff members. Currently, clonality studies are only undertaken in large specialist centres and can delay diagnosis by several weeks. Compared with immunostained tissue on a slide, WHAT IS ALREADY KNOWN ON THIS TOPIC | ⇒ T-cell lymphomas often require complex, multiplex PCR clonality tests for definitive diagnosis. We know that each alpha-beta T cell expresses either a TCRbeta1 or TCRbeta2 constant region as part of its T-cell receptor, encoded by the TRBC1 and TRBC2 gene segments, respectively.
WHAT THIS STUDY ADDS | | --- | | ⇒ This study validates a rabbit monoclonal antiTCRbeta1, rabbit monoclonal anti-TCRbeta2 and chimeric mouse monoclonal anti-TCRbeta2 antibody in formalin fixed paraffin embedded sections of T-cell lymphomas and benign T-cell populations.
HOW THIS STUDY MIGHT AFFECT RESEARCH, PRACTICE OR POLICY | ⇒ This work has the potential to make the diagnosis of T-cell lymphoma cheaper and quicker,

FAQs

Why are T-cell lymphomas difficult to diagnose compared with benign T-cell infiltrates?

T-cell lymphomas are often histologically indistinguishable from benign T-cell infiltrates. This makes diagnosis time-consuming and frequently requires clonality testing.

What approach does the study introduce to determine T-cell monotypia?

The study develops a pair of monoclonal antibodies that detect TCRβ1 and TCRβ2 in formalin-fixed, paraffin-embedded tissue. Detection is used as a surrogate for T-cell clonality/monoclonality.

How is the TCRβ2:TCRβ1 ratio obtained in the study?

Single immunostaining results are used to support automated cell counting. This enables accurate calculation of the TCRβ2:TCRβ1 ratio, including from double immunostaining examples.

Sep 23, 2026

Uploaded by Ben