Demonstration of T-Cell Monotypia Using Anti-TCRbeta1/2 (<i>TRBC1</i>/2) Immunostaining as a Rapid and Cost-Effective Alternative to PCR-Based Clonality Studies for the Diagnosis of T-Cell Lymphoma
Background/Objectives: T-cell lymphomas are often histologically indistinguishable from benign T-cell infiltrates, and diagnosis typically relies on slow, complex, and expensive multiplexed PCR reactions, requiring significant training and experience to interpret them. We aimed to raise highly speci...
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2024-11-01
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| author | Elizabeth J. Soilleux Daniel T. Rodgers Jinlong J. Situ Shelley C. Evans Venkata N. Konda Han-Chieh Yang Jianxiong Pang Isabella Gilbey Smith Pete Rajesh Maryam Salimi Soo Weei Ng Julia Jones Jodi L. Miller Rachel Etherington Margaret Ashton-Key Graham Ogg |
| author_facet | Elizabeth J. Soilleux Daniel T. Rodgers Jinlong J. Situ Shelley C. Evans Venkata N. Konda Han-Chieh Yang Jianxiong Pang Isabella Gilbey Smith Pete Rajesh Maryam Salimi Soo Weei Ng Julia Jones Jodi L. Miller Rachel Etherington Margaret Ashton-Key Graham Ogg |
| author_sort | Elizabeth J. Soilleux |
| collection | DOAJ |
| description | Background/Objectives: T-cell lymphomas are often histologically indistinguishable from benign T-cell infiltrates, and diagnosis typically relies on slow, complex, and expensive multiplexed PCR reactions, requiring significant training and experience to interpret them. We aimed to raise highly specific antibodies against the two alternatively used and very similar T-cell receptor beta constant regions, TCRbeta1 and TCRbeta2, encoded by the <i>TRBC1</i> and <i>TRBC2</i> gene segments, respectively. We sought to demonstrate the feasibility of detecting TCRbeta1 and TCRbeta2 immunohistochemically in routine clinical (formalin-fixed, paraffin-embedded (FFPE)) tissue sections as a novel diagnostic strategy for T-cell lymphomas. Methods: Recombinant rabbit antibodies were validated using Western blotting and FFPE immunostaining of T-cell leukemia lines. The immunostaining of FFPE tissue containing benign and lymphomatous T-cell populations was undertaken, with corroboration by BaseScope<sup>TM</sup> high-sensitivity in situ hybridization and quantitative real-time PCR (Q-PCR). An additional Q-PCR literature review and analysis of publicly available RNAseq data was used to determine the TCRbeta2/TCRbeta1 ratio cut-off to separate benign and malignant T-cell populations. Results: Our TCRbeta1/TCRbeta2 antibody pair gave highly specific FFPE tissue staining. All benign samples analyzed (immunohistochemically, by BaseScope<sup>TM</sup>, by Q-PCR, and by RNAseq data analysis) had TCRbeta1/TCRbeta2 or <i>TRBC1/TRBC2</i> ranges well within the previously published flow cytometric benign range (TCRbeta2/TCRbeta1 = 0.18:1–5.7:1), while samples of T-cell lymphoma did not. One out of thirteen (7.7%) lymphoma samples showed some detectable TCRbeta1/TCRbeta2 protein co-expression, and 4 out of 13 (30.8%) T-cell lymphomas showed a <i>TRBC1/TRBC2</i> transcript co-expression using BaseScope<sup>TM</sup>. Conclusions: Analyzing T-cell monotypia immunohistochemically, analogous to B-cell monotypia (kappa: lambda ratio for B-cell and plasma cell neoplasms), could make the diagnosis of T-cell lymphomas cheaper, quicker, and more accurate. Larger studies are needed to validate our antibodies for clinical use. |
| format | Article |
| id | doaj-art-e6a2c6b212a24342a8501376bc0f3ffe |
| institution | OA Journals |
| issn | 2075-4418 |
| language | English |
| publishDate | 2024-11-01 |
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| series | Diagnostics |
| spelling | doaj-art-e6a2c6b212a24342a8501376bc0f3ffe2025-08-20T01:53:45ZengMDPI AGDiagnostics2075-44182024-11-011422247910.3390/diagnostics14222479Demonstration of T-Cell Monotypia Using Anti-TCRbeta1/2 (<i>TRBC1</i>/2) Immunostaining as a Rapid and Cost-Effective Alternative to PCR-Based Clonality Studies for the Diagnosis of T-Cell LymphomaElizabeth J. Soilleux0Daniel T. Rodgers1Jinlong J. Situ2Shelley C. Evans3Venkata N. Konda4Han-Chieh Yang5Jianxiong Pang6Isabella Gilbey Smith7Pete Rajesh8Maryam Salimi9Soo Weei Ng10Julia Jones11Jodi L. Miller12Rachel Etherington13Margaret Ashton-Key14Graham Ogg15Department of Pathology, University of Cambridge, Cambridge CB2 0SP, UKHuman Research Tissue Bank, Cambridge University Hospitals NHS Foundation Trust, Cambridge CB2 0QQ, UKDepartment of Pathology, University of Cambridge, Cambridge CB2 0SP, UKDepartment of Pathology, University of Cambridge, Cambridge CB2 0SP, UKDepartment of Pathology, University of Cambridge, Cambridge CB2 0SP, UKDepartment of Pathology, University of Cambridge, Cambridge CB2 0SP, UKDepartment of Pathology, University of Cambridge, Cambridge CB2 0SP, UKDepartment of Pathology, University of Cambridge, Cambridge CB2 0SP, UKDepartment of Pathology, University of Cambridge, Cambridge CB2 0SP, UKMRC Translational Immune Discovery Unit, MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UKMRC Translational Immune Discovery Unit, MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UKCancer Research UK Cambridge Institute, Li Ka Shing Centre, University of Cambridge, Cambridge CB2 0RE, UKCancer Research UK Cambridge Institute, Li Ka Shing Centre, University of Cambridge, Cambridge CB2 0RE, UKMRC Translational Immune Discovery Unit, MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UKDepartment of Cellular Pathology, University Hospital Southampton, Southampton SO16 6YD, UKMRC Translational Immune Discovery Unit, MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UKBackground/Objectives: T-cell lymphomas are often histologically indistinguishable from benign T-cell infiltrates, and diagnosis typically relies on slow, complex, and expensive multiplexed PCR reactions, requiring significant training and experience to interpret them. We aimed to raise highly specific antibodies against the two alternatively used and very similar T-cell receptor beta constant regions, TCRbeta1 and TCRbeta2, encoded by the <i>TRBC1</i> and <i>TRBC2</i> gene segments, respectively. We sought to demonstrate the feasibility of detecting TCRbeta1 and TCRbeta2 immunohistochemically in routine clinical (formalin-fixed, paraffin-embedded (FFPE)) tissue sections as a novel diagnostic strategy for T-cell lymphomas. Methods: Recombinant rabbit antibodies were validated using Western blotting and FFPE immunostaining of T-cell leukemia lines. The immunostaining of FFPE tissue containing benign and lymphomatous T-cell populations was undertaken, with corroboration by BaseScope<sup>TM</sup> high-sensitivity in situ hybridization and quantitative real-time PCR (Q-PCR). An additional Q-PCR literature review and analysis of publicly available RNAseq data was used to determine the TCRbeta2/TCRbeta1 ratio cut-off to separate benign and malignant T-cell populations. Results: Our TCRbeta1/TCRbeta2 antibody pair gave highly specific FFPE tissue staining. All benign samples analyzed (immunohistochemically, by BaseScope<sup>TM</sup>, by Q-PCR, and by RNAseq data analysis) had TCRbeta1/TCRbeta2 or <i>TRBC1/TRBC2</i> ranges well within the previously published flow cytometric benign range (TCRbeta2/TCRbeta1 = 0.18:1–5.7:1), while samples of T-cell lymphoma did not. One out of thirteen (7.7%) lymphoma samples showed some detectable TCRbeta1/TCRbeta2 protein co-expression, and 4 out of 13 (30.8%) T-cell lymphomas showed a <i>TRBC1/TRBC2</i> transcript co-expression using BaseScope<sup>TM</sup>. Conclusions: Analyzing T-cell monotypia immunohistochemically, analogous to B-cell monotypia (kappa: lambda ratio for B-cell and plasma cell neoplasms), could make the diagnosis of T-cell lymphomas cheaper, quicker, and more accurate. Larger studies are needed to validate our antibodies for clinical use.https://www.mdpi.com/2075-4418/14/22/2479lymphomamonotypiaclonalityimmunohistochemistrydiagnostic testformalin-fixed paraffin-embedded tissue |
| spellingShingle | Elizabeth J. Soilleux Daniel T. Rodgers Jinlong J. Situ Shelley C. Evans Venkata N. Konda Han-Chieh Yang Jianxiong Pang Isabella Gilbey Smith Pete Rajesh Maryam Salimi Soo Weei Ng Julia Jones Jodi L. Miller Rachel Etherington Margaret Ashton-Key Graham Ogg Demonstration of T-Cell Monotypia Using Anti-TCRbeta1/2 (<i>TRBC1</i>/2) Immunostaining as a Rapid and Cost-Effective Alternative to PCR-Based Clonality Studies for the Diagnosis of T-Cell Lymphoma Diagnostics lymphoma monotypia clonality immunohistochemistry diagnostic test formalin-fixed paraffin-embedded tissue |
| title | Demonstration of T-Cell Monotypia Using Anti-TCRbeta1/2 (<i>TRBC1</i>/2) Immunostaining as a Rapid and Cost-Effective Alternative to PCR-Based Clonality Studies for the Diagnosis of T-Cell Lymphoma |
| title_full | Demonstration of T-Cell Monotypia Using Anti-TCRbeta1/2 (<i>TRBC1</i>/2) Immunostaining as a Rapid and Cost-Effective Alternative to PCR-Based Clonality Studies for the Diagnosis of T-Cell Lymphoma |
| title_fullStr | Demonstration of T-Cell Monotypia Using Anti-TCRbeta1/2 (<i>TRBC1</i>/2) Immunostaining as a Rapid and Cost-Effective Alternative to PCR-Based Clonality Studies for the Diagnosis of T-Cell Lymphoma |
| title_full_unstemmed | Demonstration of T-Cell Monotypia Using Anti-TCRbeta1/2 (<i>TRBC1</i>/2) Immunostaining as a Rapid and Cost-Effective Alternative to PCR-Based Clonality Studies for the Diagnosis of T-Cell Lymphoma |
| title_short | Demonstration of T-Cell Monotypia Using Anti-TCRbeta1/2 (<i>TRBC1</i>/2) Immunostaining as a Rapid and Cost-Effective Alternative to PCR-Based Clonality Studies for the Diagnosis of T-Cell Lymphoma |
| title_sort | demonstration of t cell monotypia using anti tcrbeta1 2 i trbc1 i 2 immunostaining as a rapid and cost effective alternative to pcr based clonality studies for the diagnosis of t cell lymphoma |
| topic | lymphoma monotypia clonality immunohistochemistry diagnostic test formalin-fixed paraffin-embedded tissue |
| url | https://www.mdpi.com/2075-4418/14/22/2479 |
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