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      Scientist examining test tube in a laboratory.

       

      For Professionals in Clinical Diagnostics

       

      Lymphoma affects the lymphatic system and B and T cell populations of white blood cells (WBCs). Lymphoma develops with the malignant transformation of lymphocytes. Most lymphomas originate from B cells with only 10–15% being of T and NK cell origins.1

       

       

      What causes lymphoma?

      As with many cancers, genetic alterations have been identified in several types of lymphoma and family history of the disease accounts for 5% of cases. Several factors are considered as risks promoting the development of lymphoma. These include radiation therapy, immune deficiency and viral infections such as from human immunodeficiency virus (HIV), human T lymphotropic virus type 1 (HTLV-1), hepatitis C virus (HCV) and Epstein-Barr virus (EBV).2

      Types of lymphoma

      More than 70 kinds of lymphomas have been described, and they are grouped in two main types—Hodgkin and non-Hodgkin lymphoma.

       

      Hodgkin lymphoma

      In most cases of Hodgkin lymphoma (HL), the neoplastic cells are derived from mature B-cells. It preferentially develops in young adults between 20 and 34 years old. Hodgkin Reed-Sternberg (HRS) cells are a hallmark of Hodgkin lymphoma. They are giant multinucleated cells forming a clonal tumor pool of Hodgkin lymphoma. CD30 is the hallmark of HL and HRS cell surface markers.3

       

      Non-Hodgkin lymphoma

      Non-Hodgkin lymphoma (NHL) is the most common form of lymphoma.4 It preferentially develops in older adults and is less responsive to treatment than Hodgkin lymphoma. Surface markers such as CD20 and CD19 are expressed in NHL that are derived from B lymphocytes.5 Flow cytometry–based immunophenotypic analysis of NHL relies on the detection of specific lineage markers of B and T cells.

      Flow cytometry–based assays for lymphoma

      Immunophenotyping by flow cytometry is a solution to determine the type of lymphoma as panels of markers can be run to track the aberrant expression of antigens on lymphocytes.

       

      Flow cytometry can also support lymphoma subtype diagnosis such as diffuse large B cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), Burkitt lymphoma (BL) and lymphoplasmacytic lymphoma (LPL).6 Circulating tumor cells can be detected in peripheral blood by highly sensitive flow cytometry assays at diagnosis but also serve as a prognostic marker of progression.7

       

      Immunophenotyping

      When a lymphoproliferative disorder remains a significant possibility after clinical evaluation, cell surface and some intracellular phenotyping of lymphocytes is warranted.

       

      Flow cytometry allows the screening of peripheral blood mononuclear cells (PBMCs), bone marrow and other fluids such as cerebrospinal fluid (CSF) to monitor not only the percentage of lymphocytes positive for a specific antigen but also the density of the antigens. Normal PBMC composition includes approximately 10% B cells, 80% T cells and 10% NK cells. Variations in these cell populations are cause for suspicion of lymphoproliferative disease. Specific flow cytometry panels can be run to assess each of these populations. Flow cytometry can also be used for phenotyping other lymphoid cells and tissues, including lymph nodes, skin, organs and brain as well as body fluids.  

       

      Diffuse large B cell lymphoma

      Diffuse large B cell lymphoma (DLBCL) is the largest category of lymphoma and accounts for 30 to 40% of adult NHL in western countries. This category of lymphoma can be a progression from another lymphoid malignancy such as follicular lymphoma (FL) or chronic lymphocytic leukemia (CLL). It is characterized by large lymphocytes that can be detected by flow cytometry. A standard immunophenotyping panel for DLBCL includes pan-B cell markers (CD19, CD22, CD79a), germinal center marker CD10, and proliferation marker Ki67. A high Ki67 indicates increased proliferation of the malignant lymphoma cells.8,9

       

      T cell/NK cell neoplasms

      Abnormalities in pan-T cell antigens, e.g., CD2, CD3, CD5, CD7, are indicative of a T cell lymphoproliferative disorder.10 T cell non-Hodgkin lymphoma (T cell NHL) represents about 12% of all non-Hodgkin lymphomas.11

       

      T/NK cell neoplasms represent a small portion of lymphomas. They display an aberrant T cell or NK cell phenotype. Chronic lymphoproliferative disorders of NK cells (CLPD-NK) are rare heterogeneous disorders with continuous expansion of NK cells in peripheral blood, expressing CD16 and low levels of CD56 and CD57.12 Peripheral T cell lymphoma is an uncommon heterogeneous malignancy, accounting for less than 15% of all lymphomas.13  

       

      Flow cytometry–based diagnostic applications

      BD Biosciences offers a large portfolio of single-color antibody reagents that span across a range of specificities and dyes that can help in the characterization of hematologic neoplasia. These panels are to be verified and validated by the lab.

       

      Furthermore, BD Biosciences provides the BD OneFlow™ Solution, comprising a comprehensive set of reagents, setup beads, protocols and assay templates, to help standardize leukaemia and lymphoma immunophenotyping. This can improve laboratory efficiency and enable reliability and accuracy of results.14,15

      References

      1. Jiang M, Bennani MN, Feldman AL. Lymphoma classification  update: T-cell lymphomas, Hodgkin lymphomas and histiocytic/dendritic cell neoplasms. Expert Rev Hematol. 2017;10(3):239-249. doi: 10.1080/17474086.2017.1281122

      2. Morton LM, Slager SL, Cerhan JR, et al. Etiologic heterogeneity among non-Hodgkin lymphoma subtypes: the InterLymph Non-Hodgkin Lymphoma Subtypes Project. J Natl Cancer Inst Monogr. 2014;2014(48):130-144. doi:10.1093/jncimonographs/lgu013

      3. Küppers R, Hansmann ML. The Hodgkin and Reed/Sternberg cell. Int J Biochem Cell Biol. 2005;37(3):511-517. doi:10.1016/j.biocel.2003.10.025

      4. Swerdlow SH, Campo E, Pileri SA, et al. The 2016 revision of the World Health Organization classification of lymphoid neoplasms. Blood. 2016;127(20):2375-2390. doi:10.1182/blood-2016-01-643569 

      6. Sarkozy C, Baseggio L, Feugier P, et al. Peripheral blood involvement in patients with follicular lymphoma: a rare disease manifestation associated with poor prognosis. Br J Haematol. 2014;164(5):659-667. doi:10.1111/bjh.12675

      8. van Dongen JJ, Lhermitte L, Böttcher S, et al. EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. Leukaemia. 2012;26(9):1908-1975. doi:10.1038/leu.2012.120

      10. Aujla A, Aujla R, Liu D. Inotuzumab ozogamicin in clinical development for acute lymphoblastic leukemia and non-Hodgkin lymphoma. Biomark Res. 2019;7:9. doi:10.1186/s40364-019-0160-4

      12. Abuelgasim KA, Rehan H, Alsubaie M, et al. Coexistence of chronic myeloid leukemia and diffuse large B-cell lymphoma with antecedent chronic lymphocytic leukemia: a case report and review of the literature. J Med Case Rep. 2018;12(1):64. doi:10.1186/s13256-018-1612-4

      14. Friedberg JW, Fisher RI. Diffuse large B-cell lymphoma. Hematol Oncol Clin North Am. 2008;22(5):941-ix. doi:10.1016/j.hoc.2008.07.002

      16. Pulitzer M. Cutaneous T-cell lymphoma. Clin Lab Med. 2017;37(3):527-546. doi:10.1016/j.cll.2017.06.006

      18. Park HS, McIntosh L, Braschi-Amirfarzan M, Shinagare AB, Krajewski KM. T-cell non-Hodgkin lymphomas: spectrum of disease and the role of imaging in the management of common subtypes. Korean J Radiol. 2017;18(1):71-83. doi:10.3348/kjr.2017.18.1.71

      20. Zambello R, Teramo A, Barila G et al. Activating KIRs in chronic lymphoproliferative disorder of NK cells: Protection from viruses and disease induction? Front Immunol. 2014;5:72. doi: 10.3389/fimmu.2014.00072

      22. D’Amore F, Relander T, Lauritzsen GF, et al. Up-front autologous stem-cell transplantation in peripheral T-cell lymphoma: NLG-T-01. J Clin Oncol. 2012;30(25):3093-3099. doi: 10.1200/JCO.2011.40.2719

      24. van der Velden VHJ, Flores-Montero J, Perez-Andres M, et al. Optimization and testing of dried antibody tube: The EuroFlow LST and PIDOT tubes as examples. J Immunol Methods. 2019;475:112287. doi: 10.1016/j.jim.2017.03.011

      26. Moloney E, Watson H, Barge D, et al. Efficiency and health economic evaluations of BD OneFlow™ Flow Cytometry Reagents for diagnosing chronic lymphoid leukemia. Cytometry B Clin Cytom. 2019;96(6):514-520. doi: 10.1002/cyto.b.21779

      BD Biosciences clinical flow cytometry solutions, including instrumentation, software and reagents, offer the building blocks for laboratory-developed tests used in the identification of markers associated with lymphomas.

      These solutions are not FDA cleared or approved for the diagnosis of lymphomas. Analyte Specific Reagent. Analytical and performance characteristics are not established.

      These solutions are ordered as Analyte Specific Reagents (ASR) and their analytical and performance characteristics are not established.

      Class 1 Laser Product per IEC standard

      Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

      Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

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