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    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 one of the most common forms of lymphoma.4 It preferentially develops in older adults and is less responsive to treatment than Hodgkin lymphoma.

    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. 

     

    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 CE-IVD reagents that span across a range of specificities and dyes that can help in the characterization of hematologic neoplasia. Multicolor analysis using relevant combinations of reagents is highly recommended.14

     

    Furthermore, BD Biosciences provides the BD OneFlow™ Solution, comprising a comprehensive set of reagents, setup beads, protocols and assay templates, to help standardize leukemia and lymphoma immunophenotyping. This can improve laboratory efficiency and enable reliability and accuracy of results.15,16

    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

    5. 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

    6. 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

    7. Galimberti S, Genuardi E et al. The Minimal Residual Disease in Non-Hodgkin's lymphomas: From the Laboratory to the Clinical Practice. G Front Oncol. 2019; 9: 528. doi: 10.3389/fonc.2019.00528

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

    9. 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

    10. 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

    11. 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

    12. Stelzer GT, Marti G, Hurley A, McCoy P Jr, Lovett EJ, Schwartz A. US-Canadian Consensus recommendations on the immunophenotypic analysis of hematologic neoplasia by flow cytometry: standardization and validation of laboratory procedures. Cytometry. 1997;30:214-230.

    13. 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

    14. 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 Single-Color Antibody reagents, as intended on this page, and BD OneFlow™ Reagents are in vitro diagnostic medical devices bearing a CE mark.

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