Abstract
The BIOMED-2 Concerted Action BMH4-CT98-3936 on ‘Polymerase chain reaction (PCR)-based clonality studies for early diagnosis of lymphoproliferative disorders’ developed standardized PCR protocols for detection of immunoglobulin (Ig) and T-cell receptor (TCR) rearrangements, including TCR beta (TCRB). As no comparable TCRB PCR method pre-existed and only a limited number of samples was tested within the BIOMED-2 study, we initiated this study for further validation of the newly developed TCRB PCR approach by comparing PCR data with previously generated Southern blot (SB) data in a series of 66 immature (ALL) and 36 mature T-cell malignancies. In 91% of cases, concordant PCR and SB results were found. Discrepancies consisted of either failure to detect SB-detected TCRB rearrangements by PCR (6.5%) or detection of an additional non-SB defined rearrangement (2.5%). In 99% of cases (99/100), at least one clonal TCRB rearrangement was detected by PCR in the SB-positive cases. A predominance of complete Vβ-Jβ rearrangements was seen in TCRαβ+ T-cell malignancies and CD3-negative T-ALL (100 and 90%, respectively), whereas in TCRγδ+ T-ALL, more incomplete Dβ-Jβ TCRB rearrangements were detected (73%). Our results underline the reliability of this new TCRB PCR method and its strategic applicability in clonality diagnostics of lymphoproliferative disorders and MRD studies.
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References
Van Dongen JJM, Comans-Bitter WM, Wolvers-Tettero ILM, Borst J . Development of human T lymphocytes and their thymus-dependency. Thymus 1990; 16: 207–234.
Van Dongen JJM, Wolvers-Tettero ILM . Analysis of immunoglobulin and T cell receptor genes. Part I: Basic and technical aspects. Clin Chim Acta 1991; 198: 1–91.
Van Dongen JJM, Wolvers-Tettero ILM . Analysis of immunoglobulin and T-cell receptor genes. Part II: Possibilities and limitations in the diagnosis and management of lymphoproliferative diseases and related disorders. Clin Chim Acta 1991; 198: 93–174.
Langerak AW, Wolvers-Tettero ILM, Van Dongen JJM . Detection of T cell receptor beta (TCRB) gene rearrangement patterns in T cell malignancies by Southern blot analysis. Leukemia 1999; 13: 965–974.
Breit TM, Wolvers-Tettero ILM, Beishuizen A, Verhoeven M-AJ, Van Wering ER, Van Dongen JJM . Southern blot patterns, frequencies and junctional diversity of T-cell receptor d gene rearrangements in acute lymphoblastic leukemia. Blood 1993; 82: 3063–3074.
Lefranc MP . Locus maps and genomic repertoire of the human T-cell receptor genes. The Immunologist 2000; 8: 72–79.
Lefranc MP . IMGT databases, web resources and tools for immunoglobulin and T cell receptor sequence analysis. http://imgt.cines.fr. Leukemia 2003; 17: 260–266.
Rowen L, Koop BF, Hood L . The complete 685-kilobase DNA sequence of the human beta T cell receptor locus. Science 1996; 272: 1755–1762.
Toyonaga B, Yoshikai Y, Vadasz V, Chin B, Mak TW . Organization and sequences of the diversity, joining, and constant region genes of the human T-cell receptor b chain. Proc Natl Acad Sci USA 1985; 82: 8624–8628.
Wei S, Charmley P, Robinson MA, Concannon P . The extent of the human germline T-cell receptor V beta gene segment repertoire. Immunogenetics 1994; 40: 27–36.
Tsuda S, Rieke S, Hashimoto Y, Nakauchi H, Takahama Y . Il-7 supports D-J but not V-DJ rearrangement of TCR-beta gene in fetal liver progenitor cells. J Immunol 1996; 156: 3233–3242.
van der Velden VH, Hochhaus A, Cazzaniga G, Szczepanski T, Gabert J, van Dongen JJ . Detection of minimal residual disease in hematologic malignancies by real-time quantitative PCR: principles, approaches, and laboratory aspects. Leukemia 2003; 17: 1013–1034.
Szczepanski T, van der Velden VH, Raff T, Jacobs DC, van Wering ER, Bruggemann M et al. Comparative analysis of T-cell receptor gene rearrangements at diagnosis and relapse of T-cell acute lymphoblastic leukemia (T-ALL) shows high stability of clonal markers for monitoring of minimal residual disease and reveals the occurrence of second T-ALL. Leukemia 2003; 17: 2149–2156.
Bruggemann M, Van der Velden VHJ, Raff T, Droese J, Ritgen M, Pott C et al. Rearranged TCRB genes represent powerful targets for detection of minimal residual disease by real-time quantitative PCR in adult and childhood T-ALL. Leukemia 2004; 18: 709–719.
Szczepanski T, Langerak AW, Wolvers-Tettero ILM, Ossenkoppele GJ, Verhoef G, Stul M et al. Immunoglobulin and T cell receptor gene rearrangement patterns in acute lymphoblastic leukemia are less mature in adults than in children: implications for selection of PCR targets for detection of minimal residual disease. Leukemia 1998; 12: 1081–1088.
Szczepanski T, Beishuizen A, Pongers-Willemse MJ, Hählen K, van Wering ER, Wijkhuijs JM et al. Cross-lineage T-cell receptor gene rearrangements occur in more than ninety percent of childhood precursor-B-acute lymphoblastic leukemias: alternative PCR targets for detection of minimal residual disease. Leukemia 1999; 13: 196–205.
McCarthy KP, Sloane JP, Kabarowski JH, Matutes E, Wiedemann LM . The rapid detection of clonal T-cell proliferations in patients with lymphoid disorders. Am J Pathol 1991; 138: 821–828.
Kneba M, Bolz I, Linke B, Hiddemann W . Analysis of rearranged T-cell receptor beta-chain genes by polymerase chain reaction (PCR) DNA sequencing and automated high resolution PCR fragment analysis. Blood 1995; 86: 3930–3937.
Assaf C, Hummel M, Dippel E, Goerdt S, Muller HH, Anagnostopoulos I et al. High detection rate of T-cell receptor beta chain rearrangements in T-cell lymphoproliferations by family specific polymerase chain reaction in combination with the GeneScan technique and DNA sequencing. Blood 2000; 96: 640–646.
Beers T, Du TL, Rickert M, Overturf P, Choi Y, Greenberg SJ . Ex vivo clonotype primer-directed gene amplification to identify malignant T cell repertoires. J Leukocyte Biol 1993; 54: 343–350.
Gorski J, Yassai M, Zhu X, Kissella B, Keever C, Flomenberg N . Circulating T cell repertoire complexity in normal individuals and bone marrow recipients analyzed by CDR3 size spectratyping. Correlation with immune status. J Immunol 1994; 152: 5109–5119.
O'Shea U, Wyatt JI, Howdle PD . Analysis of T cell receptor beta chain CDR3 size using RNA extracted from formalin fixed paraffin wax embedded tissue. J Clin Pathol 1997; 50: 811–814.
Van Dongen JJM, Langerak AW, Bruggemann M, Evans PAS, Hummel M, Lavender FL et al. Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: report of the BIOMED-2 Concerted Action BMH4-CT98-3936. Leukemia 2003; 17: 2257–2317.
Langerak AW, Szczepanski T, Van der Burg M, Wolvers-Tettero ILM, Van Dongen JJM . Heteroduplex PCR analysis of rearranged T cell receptor genes for clonality assessment in suspect T cell proliferations. Leukemia 1997; 11: 2192–2199.
Langerak AW, van Den Beemd R, Wolvers-Tettero IL, Boor PP, van Lochem EG, Hooijkaas H et al. Molecular and flow cytometric analysis of the Vbeta repertoire for clonality assessment in mature TCRalphabeta T-cell proliferations. Blood 2001; 98: 165–173.
Duby AD, Seidman JG . Abnormal recombination products result from aberrant DNA rearrangement of the human T-cell antigen receptor beta-chain gene. Proc Natl Acad Sci USA 1986; 83: 4890–4894.
Alatrakchi N, Farace F, Frau E, Carde P, Munck JN, Triebel F . T-cell clonal expansion in patients with B-cell lymphoproliferative disorders. J Immunother 1998; 21: 363–370.
Guidal C, Vilmer E, Grandchamp B, Cave H . A competitive PCR-based method using TCRD, TCRG and IGH rearrangements for rapid detection of patients with high levels of minimal residual disease in acute lymphoblastic leukemia. Leukemia 2002; 16: 762–764.
Foa R, Pelicci PG, Migone N, Lauria F, Pizzolo G, Flug F et al. Analysis of T-cell receptor beta chain (T beta) gene rearrangements demonstrates the monoclonal nature of T-cell chronic lymphoproliferative disorders. Blood 1986; 67: 247–250.
Feller AC, Griesser H, Schilling CV, Wacker HH, Dallenbach F, Bartels H et al. Clonal gene rearrangement patterns correlate with immunophenotype and clinical parameters in patients with angioimmunoblastic lymphadenopathy. Am J Pathol 1988; 133: 549–556.
Asnafi V, Beldjord K, Boulanger E, Comba B, Le Tutour P, Estienne MH et al. Analysis of TCR, pT alpha, and RAG-1 in T-acute lymphoblastic leukemias improves understanding of early human T-lymphoid lineage commitment. Blood 2003; 101: 2693–2703.
Acknowledgements
We gratefully acknowledge Petra Chall, Frauke Hemken, Ellen van Gastel-Mol for their technical assistance and Mr Tar van Os for help in preparation of the figure. This work was supported by the BIOMED-2 Concerted Action BMH4-CT98-3936 ‘PCR-based clonality studies for early diagnosis of lymphoproliferative disorders’ of the European Commission.
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Droese, J., Langerak, A., Groenen, P. et al. Validation of BIOMED-2 multiplex PCR tubes for detection of TCRB gene rearrangements in T-cell malignancies. Leukemia 18, 1531–1538 (2004). https://doi.org/10.1038/sj.leu.2403428
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DOI: https://doi.org/10.1038/sj.leu.2403428
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