Diagnostic Value of Genotypic Analysis in Primary Cutaneous Lymphomas using Standardized BIOMED-2 Polymerase Chain Reaction Protocols: Experience in Daily Clinical Practice
DOI:
https://doi.org/10.2340/00015555-3828Keywords:
BIOMED-2, clonality, cutaneous B-cell lymphoma, cutaneous T-cell lymphoma, multiplex polymerase chain reaction, T-cell receptor genes, immunoglobulin genesAbstract
BIOMED-2 Concerted Action BMH4-CT98-3936 (BIOMED-2) PCR protocols are an important diagnostic tool in the evaluation of cutaneous lymphomas. The aim of this study was to assess the diagnostic value of the genotyping results obtained by these techniques in daily clinical practice. A total of 360 paraffin-embedded skin samples were retrospectively reviewed from 114 cutaneous T-cell lymphomas and 35 cutaneous B-cell lymphomas. A total of 249 biopsies from 180 patients with benign lymphoid infiltrates served as controls. T-cell receptor and immunoglobulin gene rearrangements were assessed using the BIOMED-2 method. A combined T-cell receptor gamma and beta assay approach reliably distinguished cutaneous T-cell lymphomas from benign skin T-cell infiltrates (sensitivity 89.4%; specificity 81.5%). Analysis of complete immunoglobulin heavy chain rearrangements also differentiated cutaneous B-cell lymphomas from benign B-cell infiltrates (sensitivity 85.7%; specificity 82.4%). In conclusion, the full BIOMED-2 protocol is a useful aid combined with clinical, histological and immunophenotypical findings for assessment of lymphoid clonality in skin lymphoid proliferations.
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Willemze R, Jaffe ES, Burg G, Cerroni L, Berti E, Swerdlow SH, et al. WHO-EORTC classification for cutaneous lymphomas. Blood 2005; 105: 3768-3785.
DOI: https://doi.org/10.1182/blood-2004-09-3502
Willemze R, Cerroni L, Kempf W, Berti E, Facchetti F, Swerdlow SH, et al. The 2018 update of the WHO-EORTC classification for primary cutaneous lymphomas. Blood 2019; 133: 1703-1714.
DOI: https://doi.org/10.1182/blood-2018-11-881268
Goeldel AL, Cornillet-Lefebvre P, Durlach A, Birembaut P, Bernard P, Nguyen P, et al. T-cell receptor ? gene rearrangement in cutaneous T-cell lymphoma: comparative study of polymerase chain reaction with denaturing gradient gel electrophoresis and GeneScan analysis. Br J Dermatol 2009; 162: 822-829.
DOI: https://doi.org/10.1111/j.1365-2133.2009.09575.x
Lukowsky A, Muche JM, Möbs M, Assaf C, Humme D, Hummel M, et al. Evaluation of T-cell clonality in archival skin biopsy samples of cutaneous T-cell lymphomas using the Biomed-2 PCR protocol. Diagn Mol Pathol 2010; 19: 70-77.
DOI: https://doi.org/10.1097/PDM.0b013e3181b2a1b7
Felcht M, Booken N, Stroebel P, Goerdt S, Klemke CD. The value of molecular diagnostics in primary cutaneous B-cell lymphomas in the context of clinical findings, histology, and immunohistochemistry. J Am Acad Dermatol 2011; 64: 135-143.
DOI: https://doi.org/10.1016/j.jaad.2009.12.062
Bouloc A, Delfau-Larue MH, Lenormand B, Meunier F, Wechsler J, Thomine E, et al. Polymerase chain reaction analysis of immunoglobulin gene rearrangement in cutaneous lymphoid hyperplasias. Arch Dermatol 1999; 135: 168-172.
DOI: https://doi.org/10.1001/archderm.135.2.168
Sandberg Y, Heule F, Lam K, Lugtenburg PJ, Wolvers-Tettero IL, van Dongen JJ, et al. Molecular immunoglobulin/T-cell receptor clonality analysis in cutaneous lymphoproliferations. Experience with the BIOMED-2 standardized polymerase chain reaction protocol. Haematologica 2003; 88: 659-670.
Zhang B, Beck AH, Taube JM, Kohler S, Seo K, Zwerner J, et al. Combined use of PCR-based TCRG and TCRB clonality tests on paraffin-embedded skin tissue in the differential diagnosis of mycosis fungoides and inflammatory dermatoses. J Mol Diagn 2010; 12: 320-327.
DOI: https://doi.org/10.2353/jmoldx.2010.090123
Morales AV, Arber DA, Seo K, Kohler S, Kim YH, Sundram UN. Evaluation of B-cell clonality using the BIOMED-2 PCR method effectively distinguishes cutaneous B-cell lymphoma from benign lymphoid infiltrates. Am J Dermatopathol 2008; 30: 425-430.
DOI: https://doi.org/10.1097/DAD.0b013e31818118f7
Guitart J, Magro C. Cutaneous T-cell lymphoid dyscrasia: a unifying term for idiopathic chronic dermatoses with persistent T-cell clones. Arch Dermatol 2007; 143: 921-932.
DOI: https://doi.org/10.1001/archderm.143.7.921
Lukowsky A, Muche JM, Sterry W, Audring H. Detection of expanded T cell clones in skin biopsy samples of patients with lichen sclerosus et atrophicus by T cell receptor-? polymerase chain reaction assays. J Invest Dermatol 2000; 115: 254-259.
DOI: https://doi.org/10.1046/j.1523-1747.2000.00040.x
Magro C, Crowson AN, Kovatich A, Burns F. Pityriasis lichenoides: a clonal T-cell lymphoproliferative disorder. Hum Pathol 2002; 33: 788-795.
DOI: https://doi.org/10.1053/hupa.2002.125381
Ponti R, Quaglino P, Novelli M, Fierro MT, Comessatti A, Peroni A, et al. T-cell receptor gamma gene rearrangement by multiplex polymerase chain reaction/heteroduplex analysis in patients with cutaneous T-cell lymphoma (mycosis fungoides/Sézary syndrome) and benign inflammatory disease: correlation with clinical, histological and immunophenotypical findings. Br J Dermatol 2005; 153: 565-573.
DOI: https://doi.org/10.1111/j.1365-2133.2005.06649.x
Möbs M, Cerroni L, Flaig MJ, Lenze D, Hummel M, Assaf C. Molecular diagnostics in cutaneous lymphomas. J Dtsch Dermatol Ges 2013; 11: 25-35.
DOI: https://doi.org/10.1111/ddg.12084_supp
van Dongen JJ, Langerak AW, Brüggemann M, Evans PA, 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.
DOI: https://doi.org/10.1038/sj.leu.2403202
Pimpinelli N, Olsen EA, Santucci M, Vonderheid E, Haeffner AC, Stevens S, et al. Defining early mycosis fungoides. J Am Acad Dermatol 2005; 53: 1053-1063.
DOI: https://doi.org/10.1016/j.jaad.2005.08.057
Langerak AW, Groenen PJ, Brüggemann M, Beldjord K, Bellan C, Bonello L, et al. EuroClonality/BIOMED-2 guidelines for interpretation and reporting of Ig/TCR clonality testing in suspected lymphoproliferations. Leukemia 2012; 26: 2159-2171.
DOI: https://doi.org/10.1038/leu.2012.246
Stock C, Hielscher T. Package 'DTComPair' comparison of binary diagnostic tests in a paired study design. 2015 [cited 2020 Jun 21]. Available from: https://cran.r-project.org/web/packages/DTComPair/DTComPair.pdf.
Cozzio A, French LE. T-cell clonality assays: How do they compare? J Invest Dermatol 2008; 128: 771-773.
DOI: https://doi.org/10.1038/jid.2008.49
Schachter O, Tabibian-Keissar H, Debby A, Segal O, Baum S, Barzilai A. Evaluation of the polymerase chain reaction-based T-cell receptor ? clonality test in the diagnosis of early mycosis fungoides. J Am Acad Dermatol 2020; 83: 1400-1405.
DOI: https://doi.org/10.1016/j.jaad.2020.05.110
Thurber SE, Zhang B, Kim YH, Schrijver I, Zehnder J, Kohler S. T-cell clonality analysis in biopsy specimens from two different skin sites shows high specificity in the diagnosis of patients with suggested mycosis fungoides. J Am Acad Dermatol 2007; 57: 782-790.
DOI: https://doi.org/10.1016/j.jaad.2007.06.004
Mitteldorf C, Kempf W. Cutaneous pseudolymphoma-a review on the spectrum and a proposal for a new classification. J Cutan Pathol 2020; 47: 76-97.
DOI: https://doi.org/10.1111/cup.13532
Lukowsky A, Marchwat M, Sterry W, Gellrich S. Evaluation of B-cell clonality in archival skin biopsy samples of cutaneous B-cell lymphoma by immunoglobulin heavy chain gene polymerase chain reaction. Leuk Lymphoma 2006; 47: 487-493.
DOI: https://doi.org/10.1080/10428190500305380
Fujiwara M, Morales AV, Seo K, Kim YH, Arber DA, Sundram UN. Clonal identity and differences in primary cutaneous B-cell lymphoma occurring at different sites or time points in the same patient. Am J Dermatopathol 2013; 35: 11-18.
DOI: https://doi.org/10.1097/DAD.0b013e318255dbae
Schafernak KT, Variakojis D, Goolsby CL, Tucker RM, Martínez-Escala ME, Smith FA, et al. Clonality assessment of cutaneous B-cell lymphoid proliferations: a comparison of flow cytometry immunophenotyping, molecular studies, and immunohistochemistry/in situ hybridization and review of the literature. Am J Dermatopathol 2014; 36: 781-795.
DOI: https://doi.org/10.1097/DAD.0000000000000022
Nihal M, Mikkola D, Horvath N, Gilliam AC, Stevens SR, Spiro TP, et al. Cutaneous lymphoid hyperplasia: a lymphoproliferative continuum with lymphomatous potential. Hum Pathol 2003; 34: 617-622.
DOI: https://doi.org/10.1016/S0046-8177(03)00075-3
Muche JM, Lukowsky A, Asadullah K, Gellrich S, Sterry W. Demonstration of frequent occurrence of clonal T cells in the peripheral blood of patients with primary cutaneous T-cell lymphoma. Blood 1997; 90: 1636-1642.
DOI: https://doi.org/10.1182/blood.V90.4.1636.1636_1636_1642
Delfau-Larue MH, Laroche L, Wechsler J, Lepage E, Lahet C, Asso-Bonnet M, et al. Diagnostic value of dominant T-cell clones in peripheral blood in 363 patients presenting consecutively with a clinical suspicion of cutaneous lymphoma. Blood 2000; 96: 2987-2992.
DOI: https://doi.org/10.1182/blood.V96.9.2987
Muche JM, Sterry W, Gellrich S, Rzany B, Audring H, Lukowsky A. Peripheral blood T-cell clonality in mycosis fungoides and nonlymphoma controls. Diagn Mol Pathol 2003; 12: 142-150.
DOI: https://doi.org/10.1097/00019606-200309000-00005
Russell-Jones R, Whittaker S. T-cell receptor gene analysis in the diagnosis of Sezary syndrome. J Am Acad Dermatol 1999; 41: 254-259.
DOI: https://doi.org/10.1016/S0190-9622(99)70057-5
Cordel N, Lenormand B, Courville P, Helot MF, Benichou J, Joly P. Usefulness of cutaneous T-cell clonality analysis for the diagnosis of cutaneous T-cell lymphoma in patients with erythroderma. Arch Pathol Lab Med 2005; 129: 372-376.
DOI: https://doi.org/10.5858/2005-129-372-UOCTCA
Fraser-Andrews EA, Woolford AJ, Russell-Jones R, Seed PT, Whittaker SJ. Detection of a peripheral blood T cell clone is an independent prognostic marker in mycosis fungoides. J Invest Dermatol 2000; 114: 117-121.
DOI: https://doi.org/10.1046/j.1523-1747.2000.00830.x
Muche JM, Lukowsky A, Ahnhudt C, Gellrich S, Sterry W. Peripheral blood T cell clonality in mycosis fungoides - an independent prognostic marker? J Invest Dermatol 2000; 115: 504-505.
DOI: https://doi.org/10.1046/j.1523-1747.2000.00088.x
Beylot-Barry M, Sibaud V, Thiebaut R, Vergier B, Beylot C, Delaunay M, et al. Evidence that an identical T cell clone in skin and peripheral blood lymphocytes is an independent prognostic factor in primary cutaneous T cell lymphomas. J Invest Dermatol 2001; 117: 920-926.
DOI: https://doi.org/10.1046/j.0022-202x.2001.01476.x
Iyer A, Hennessey D, O'Keefe S, Patterson J, Wang W, Wong GK, et al. Skin colonization by circulating neoplastic clones in cutaneous T-cell lymphoma. Blood 2019; 134: 1517-1527.
DOI: https://doi.org/10.1182/blood.2019002516
Scarisbrick JJ, Hodak E, Bagot M, Stranzenbach R, Stadler R, Ortiz-Romero PL, et al. Developments in the understanding of blood involvement and stage in mycosis fungoides/Sezary syndrome. Eur J Cancer 2018; 101: 278-280.
DOI: https://doi.org/10.1016/j.ejca.2018.06.013
Legrand A, Pham-Ledard A, Menguy S, Vergier B, Dubus P, Merlio JP, et al. Lack of clinical relevance of blood clonality in primary cutaneous marginal zone B-cell lymphoma. Eur J Dermatol 2021; 31: 94-96.
DOI: https://doi.org/10.1684/ejd.2020.3955
Sufficool KE, Lockwood CM, Abel HJ, Hagemann IS, Schumacher JA, Kelley TW, et al. T-cell clonality assessment by next-generation sequencing improves detection sensitivity in mycosis fungoides. J Am Acad Dermatol 2015; 73: 228-236.e2.
DOI: https://doi.org/10.1016/j.jaad.2015.04.030
Kirsch IR, Watanabe R, O'Malley JT, Williamson DW, Scott LL, Elco CP, et al. TCR sequencing facilitates diagnosis and identifies mature T cells as the cell of origin in CTCL. Sci Transl Med 2015; 7: 308ra158.
DOI: https://doi.org/10.1126/scitranslmed.aaa9122
Matos TR, de Rie MA, Teunissen MBM. Research techniques made simple: high-throughput sequencing of the T-cell receptor. J Invest Dermatol 2017; 137: e131-e138.
DOI: https://doi.org/10.1016/j.jid.2017.04.001
Rea B, Haun P, Emerson R, Vignali M, Farooqi M, Samimi S, et al. Role of high-throughput sequencing in the diagnosis of cutaneous T-cell lymphoma. J Clin Pathol 2018; 71: 814-820.
DOI: https://doi.org/10.1136/jclinpath-2018-205004
De Masson A, O'Malley JT, Elco CP, Garcia SS, Divito SJ, Lowry EL, et al. High-throughput sequencing of the T cell receptor ? gene identifies aggressive early-stage mycosis fungoides. Sci Transl Med 2018; 10: eaar5894.
DOI: https://doi.org/10.1126/scitranslmed.aar5894
Wang J, Rea B, Haun P, Emerson R, Kirsch I, Bagg A. High-throughput sequencing of the T-cell receptor ? chain gene distinguishes 2 subgroups of cutaneous T-cell lymphoma. J Am Acad Dermatol 2019; 80: 1148-1150.e1.
DOI: https://doi.org/10.1016/j.jaad.2018.10.027
Ruggiero E, Nicolay JP, Fronza R, Arens A, Paruzynski A, Nowrouzi A, et al. High-resolution analysis of the human T-cell receptor repertoire. Nat Commun 2015; 6: 8081.
DOI: https://doi.org/10.1038/ncomms9081
Olsen E, Vonderheid E, Pimpinelli N, Willemze R, Kim Y, Knobler R, et al. Revisions to the staging and classification of mycosis fungoides and Sézary syndrome: a proposal of the International Society for Cutaneous Lymphomas (ISCL) and the cutaneous lymphoma task force of the European Organization of Research and Treatment of Cancer. Blood 2007; 110: 1713-1722.
DOI: https://doi.org/10.1182/blood-2007-03-055749
Kim YH, Willemze R, Pimpinelli N, Whittaker S, Olsen EA, Ranki A, et al. TNM classification system for primary cutaneous lymphomas other than mycosis fungoides and Sézary syndrome: a proposal of the International Society for Cutaneous Lymphomas (ISCL) and the Cutaneous Lymphoma Task Force of the European Organization of Research and Treatment of Cancer (EORTC). Blood 2007; 110: 479-484.
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