Abstract
Diffuse large B-cell lymphoma (DLBCL) is a fast growing and aggressive high grade non-Hodgkin lymphoma (HG-NHL) that spreads quickly, present with lymphadenopathy or enlarged nodes in mediastinum, mesenteric region or peritoneum, from where tissue excision and biopsy with histopathology is not possible. The aim of this study was to diagnose DLBCL by Flow cytometric immunophenotyping (FCI) on fine needle aspirate (FNA) of lymphnode following immunophenotypic diagnostic criteria based on expression of CD markers. All samples were preliminary assessed by fine needle aspiration cytology (FNAC) as NHL or lymph proliferative disorder (LPD). FCI was performed with a complete panel of antibodies (CD3, CD4, CD8, CD5, CD7, CD10, CD19, CD20, CD23, CD22, CD25, CD30, CD45, CD79a, CD79b, CD95, CD56, FMC7, CD40, CD15, Kappa, Lambda and Bcl-2) by dual color flowcytometry. FCI data were interpreted to diagnose and sub classify NHL according to WHO classification. Wherever possible the diagnoses were compared with available immunohistochemistry (IHC) and histopathology reports. During one year period (from February 2016 to March 2017)10 cases of DLBCL were identified by FCI. Out of 10 DLBCL cases, 8 histopathology and 6 1HC reports were available of which 6 DLBCL cases showed 100% (6/6) concordance with combined histopathology and IHC and 100% concordance with IHC alone; 75% (6/8) concordance and 25% (2/8) discordance with histopathology alone. Ig light chain was detected in 8 (80%) DLBCL cases and 40% (4/10) DLBCL cases showed Bcl-2 expression. Diagnosis of DLBCL by FCI on FNA of Lymph node can be of great help as in most cases of DLBCL biopsy of histopathology is not possible. Detection of Bcl-2 expression can help to assess the prognosis of the disease and resistance to chemotherapy.
Keywords: Flow ytometry, Immunophenotyping, Cytology, DLBCL, HG-NHL, Bcl-2.
References
- Jaffe ES, Harris NL, Stein H, Campo E, Pileri SA, Swerdlow SH. Introduction and overview of the classification of the lymphoid neoplasms.WHO classification oftumours of haematopoietic and lymphoid tissues 2008;158-166.
- Swerdlow SH, CampoE, Pileri SA, et al The 2016 revision of the World Health Organization classification of lymphoid neoplasms. Blood 2016;127(20): 2375-2390.
- John NA, Slack GW, Savage KJ, Et al. Concurrent expression of MYC and BCL 2 in diffuse large B-Cell lymphoma treated with rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisolone. J Clinoncol 2012; 30 (28): 3452-3459.
- Perry AM, crockett D, Dave BJ, et al. B-Cell lymphoma, unclassifiable, with features intermediate between diffuse large B-Cell lymphoma and burkitt lymphoma: study of 39 cases. Br J Haematol 2013; 162(I): 40-49.
- McDonnel TJ, Deane N, Platt FM, et al. bcl-2 immunoglobulin trans genic mice demonstrate extended B-Cell survival and follicular lymphoproliferation. Cell 1989; 58 (I): 79-88.
- Pierreseques, Nathalie A. Johnson. Approach to the diagnosis and treatment of high grade B-Cell lymphomas with MYC and BCL 2 and / or BCL 6 rearrangement. Blood 2017; 129(3): 280-288.
- Steel BL, Schwartz MR, Ibrahim R. Fine needle aspiration biopsy in diagnosis of lymphadenopathy in 1103 patients. Acta Cytologica 1995;39:76-81.
- Braylan RC, Orfao A, Borowitz MJ, Davis BH. Optimal number of reagents required to evaluate hematolymphoidneoplasias: results of an international consensus meeting. Cytometry 2001; 46(1):23-27.
- Dunphy CH and Ramos R. Combining fine‐needle aspiration and flow lymphoma: A retrospective review. Diagnostic cytopathology 1997;16(3):200-206.
- Young NA, Al‐Saleem TI, Ehya H, Smith Utilization of fine‐needle aspiration cytology and flow cytometry in the diagnosis and sub classification of primary and recurrent lymphoma. Cancer Cytopathology 1998;84(4):252-261.
- Bangerter M, Hildebrand A, Griesshammer M. Immunophenotypic analysis of simultaneous specimens from different sites from the same patient with malignant lymphoma.Cytopathology 2001; 12(3): 168-176.
- Mather JP, RobbersPE. Introduction to cell and tissue culture:Theory and Technique .Plenum press, New York;1998:11-1
- Stetler-Stevenson M. Flow cytometry in lymphoma diagnosis and prognosis: useful ? Best Pract Res Clin Haematol 2003; 16: 583-597.
- DeMay RM. The Art and Science of Cytopathology, 2nd Chicago: ASCP Press; 2012: 319
- Mc Pherson, R.A. and Pincus, M.R., 2017. Henry's Clinical Diagnosis and Management by Laboratory Methods E-Book. Elsevier Health Sciences
- Mahmud F, 2014, ‘Establishment of Pathology Based tumor registry at BSMMU’, Department of Pathology, BSMMU January 2014.
- Parker A, Bain B, Devereux S,et al. Best practice in lymphoma diagnosis and reporting. London, UK: British Committee for Standards inHaematology, Royal College of Pathologists;2010.
- Craig FE, Foon KA. Flow cytometric immunophenotyping for hematologic neoplasms. Blood2008 ;111(8):3941-3967.
- Dey P, Amir T, Al Jassar A et al. Combined applications of fine needle aspiration cytology and Flow cytometric immunphenotyping for diagnosis and classification of Non-Hodgkin Lymphoma. Cytojournal 2006; 3(1):24-31.
- Li S, Eshleman JR, Borowitz MJ. Lack of surface immunoglobulin light chain expression by flow cytometric immunophenotyping can help diagnose peripheral B-cell lymphoma. American journal of clinical pathology 2002 ; 118(2):229-234.
- Tomita N, Takeuchi K, Hyo R, et al. Diffuse Large B-Cell Lymphoma without immunoglobulin light chain restriction by flow cytometry. Acta Haematol 2009;121: 196-201.
- Xu Y, McKenna RW, Molberg KH, Kroft SH. Clinicopathologic analysis of CD10+ and CD10–diffuse large B-cell lymphoma: identification of a high-risk subset with coexpression of CD10 and bcl-2. American journal of clinical pathology 2001; 116(2):183-190.
- Dong HY, Harris N.L,Preffer FI, Pitman MB. Fine-needle aspiration biopsy in the diagnosis and classification of primary and recurrent lymphoma: a retrospective analysis of the utility of cytomorphology and flow cytometry. Modern Pathology 2001;14(5):472-481.
- Lai R, Arber DA, Chang KI, Wilson CS, Weiss LM. Frequency of bcl-2 expression in non-Hodgkin’s lymphoma: a study of 778 cases with comparison of marginal zone lymphoma and monocytoid B-Cell hyperplasia. Mod pathol 1998; 11: 864-869.
- Cornfield DB, Mitchell DM, Almasri NM, Anderson JB, Ahrens KP, Dooley EO, Braylan RC. Follicular lymphoma can be distinguished from benign follicularhyperplasia by flow cytometry using simultaneous stainig of cytoplasmic BCL-2 and cell surface CD20. Am J clinpathol 2000; 114: 258-263.
- Cook JR, Craig FE, Swerdlow SH. bcl-2 expression by multicolor flow cytometric analysis assists in the diagnosis of follicular lymphoma in lymph node and bone marrow. Am J Clinpathol 2003; 119:145-151.
Corresponding Author
Dr Shirin Tarafder
Associate Professor, Department of Microbiology and Immunology, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh
Email: This email address is being protected from spambots. You need JavaScript enabled to view it., This email address is being protected from spambots. You need JavaScript enabled to view it., Phone: +8801732 622242
DOI: https://dx.doi.org/10.18535/jmscr/v6i9.13
