Title: Flow cytometric immunophenotyping including Bcl-2 detection on fine needle aspirates of lymph node in the diagnosis of Diffuse Large B-Cell Lymphoma

Authors: Dr Shirin Tarafder, Dr Khondoker Hafiza Khanom, Professor Humayun Sattar

 DOI:  https://dx.doi.org/10.18535/jmscr/v6i9.13

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.

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Corresponding Author

Dr Shirin Tarafder

Associate Professor, Department of Microbiology and Immunology, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh

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