Original Article

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Blood Res 2021; 56(1):

Published online March 31, 2021

https://doi.org/10.5045/br.2021.2020146

© The Korean Society of Hematology

Optimal panel of immunohistochemistry for the diagnosis of B-cell non-Hodgkin lymphoma using bone marrow biopsy: a tertiary care center study

Nisha Marwah1, Manali Satiza1, Niti Dalal1, Sudhir Atri2, Monika Gupta1, Sunita Singh1, Rajeev Sen1

Departments of 1Pathology and 2Medicine, Pt. B. D. Sharma PGIMS, Rohtak, India

Correspondence to : Niti Dalal, M.D.
Department of Pathology, Pt. B. D. Sharma, PGIMS, Rohtak, Haryana 124001, India
E-mail: drnitidalal@gmail.com

Received: June 22, 2020; Revised: December 17, 2020; Accepted: December 17, 2020

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background
Morphological diagnosis of non-Hodgkin lymphoma (NHL) is usually based on lymph node biopsy. Bone marrow biopsy (BMB) is important for staging, and morphology alone can be challenging for subtyping. Immunohistochemistry (IHC) allows a more precise diagnosis and characterization of NHL using monoclonal antibodies. However, there is a need for a minimal panel that can provide maximum information at an affordable cost.
Methods
All newly diagnosed cases of B-cell NHL with bone marrow infiltration between 2017 and 2019 were included. BMB was the primary procedure for diagnosing B-cell NHL. Subtyping of lymphomas was performed by immunophenotyping using a panel of monoclonal antibodies on IHC. The primary diagnostic panel of antibodies for B-cell NHL included CD19, CD20, CD79, CD5, CD23, CD10, Kappa, and Lambda. The extended panel of antibodies for further subtyping included CD30, CD45, CD56, Cyclin D1, BCL2, and BCL6.
Results
All cases of B-cell NHL were classified into the chronic lymphocytic leukemia (CLL) and non-CLL groups based on morphology and primary IHC panel. In the CLL group, the most significant findings were CD5 expression, CD23 expression, dim CD79 expression, and weak surface immunoglobulin (Ig) positivity. In the non-CLL group, they were CD5 expression, positive or negative CD23 expression, strong CD79 expression, and strong surface Ig expression. An extended panel was used for further subtyping of non-CLL cases, which comprised CD10, Cyclin D1, BCL2, and BCL6.
Conclusion
We propose a two-tier approach for immunophenotypic analysis of newly diagnosed B-cell NHL cases with a minimum primary panel including CD5, CD23, CD79, Kappa, and Lambda for differentiation into CLL/non-CLL group and Kappa and Lambda for clonality assessment. An extended panel may be used wherever required for further subtyping of non-CLL.

Keywords Non-Hodgkin lymphoma, Immunohistochemistry, Chronic lymphocytic leukemia, Non-chronic lymphocytic leukemia, Bone marrow biopsy

Article

Original Article

Blood Res 2021; 56(1): 26-30

Published online March 31, 2021 https://doi.org/10.5045/br.2021.2020146

Copyright © The Korean Society of Hematology.

Optimal panel of immunohistochemistry for the diagnosis of B-cell non-Hodgkin lymphoma using bone marrow biopsy: a tertiary care center study

Nisha Marwah1, Manali Satiza1, Niti Dalal1, Sudhir Atri2, Monika Gupta1, Sunita Singh1, Rajeev Sen1

Departments of 1Pathology and 2Medicine, Pt. B. D. Sharma PGIMS, Rohtak, India

Correspondence to:Niti Dalal, M.D.
Department of Pathology, Pt. B. D. Sharma, PGIMS, Rohtak, Haryana 124001, India
E-mail: drnitidalal@gmail.com

Received: June 22, 2020; Revised: December 17, 2020; Accepted: December 17, 2020

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background
Morphological diagnosis of non-Hodgkin lymphoma (NHL) is usually based on lymph node biopsy. Bone marrow biopsy (BMB) is important for staging, and morphology alone can be challenging for subtyping. Immunohistochemistry (IHC) allows a more precise diagnosis and characterization of NHL using monoclonal antibodies. However, there is a need for a minimal panel that can provide maximum information at an affordable cost.
Methods
All newly diagnosed cases of B-cell NHL with bone marrow infiltration between 2017 and 2019 were included. BMB was the primary procedure for diagnosing B-cell NHL. Subtyping of lymphomas was performed by immunophenotyping using a panel of monoclonal antibodies on IHC. The primary diagnostic panel of antibodies for B-cell NHL included CD19, CD20, CD79, CD5, CD23, CD10, Kappa, and Lambda. The extended panel of antibodies for further subtyping included CD30, CD45, CD56, Cyclin D1, BCL2, and BCL6.
Results
All cases of B-cell NHL were classified into the chronic lymphocytic leukemia (CLL) and non-CLL groups based on morphology and primary IHC panel. In the CLL group, the most significant findings were CD5 expression, CD23 expression, dim CD79 expression, and weak surface immunoglobulin (Ig) positivity. In the non-CLL group, they were CD5 expression, positive or negative CD23 expression, strong CD79 expression, and strong surface Ig expression. An extended panel was used for further subtyping of non-CLL cases, which comprised CD10, Cyclin D1, BCL2, and BCL6.
Conclusion
We propose a two-tier approach for immunophenotypic analysis of newly diagnosed B-cell NHL cases with a minimum primary panel including CD5, CD23, CD79, Kappa, and Lambda for differentiation into CLL/non-CLL group and Kappa and Lambda for clonality assessment. An extended panel may be used wherever required for further subtyping of non-CLL.

Keywords: Non-Hodgkin lymphoma, Immunohistochemistry, Chronic lymphocytic leukemia, Non-chronic lymphocytic leukemia, Bone marrow biopsy

Fig 1.

Figure 1.Immunohistochemical profile of chronic lymphocytic leukemia showing CD19+, CD20+, CD5+, CD23+, CD79-, non-specific Lambda (immunohistochemistry, ×400).
Blood Research 2021; 56: 26-30https://doi.org/10.5045/br.2021.2020146

Fig 2.

Figure 2.Immunohistochemical profile of mantle cell lymphoma showing CD5+, CD79+, Cyclin D1+, CD23- (immunohistochemistry, ×400).
Blood Research 2021; 56: 26-30https://doi.org/10.5045/br.2021.2020146

Fig 3.

Figure 3.Immunohistochemical profile of follicular lymphoma showing CD19+ in the lymphoid follicle, CD10+ in the lymphoid follicle, BCL2+ in the lymphoid follicle, CD5-B cells in the lymphoid follicle with CD5+ scattered T-cells in the background (immunohisto-chemistry, ×400).
Blood Research 2021; 56: 26-30https://doi.org/10.5045/br.2021.2020146

Table 1 . Immunophenotypic diagnostic criteria [9]..

CD19CD20CD23CD5CD79CD10BCL2BCL6Cyclin D1
CLL++++Dim+----
NHL (unclassified)+++++----
MCL++-++---+
FL++--+++--
DLBCL++-+++-+-

Abbreviations: CLL, chronic lymphocytic leukemia; DLBCL, diffuse large B-cell lymphoma; FL, follicular lymphoma; MCL, mantle cell lymphoma; NHL, non-Hodgkin lymphoma..


Table 2 . Distribution of cases on the basis of bone marrow aspiration findings..

BMA findingsN of cases%
Chronic lymphoproliferative disorder1963.3
Chronic lymphocytic leukemia413.3
Atypical lymphocytosis516.6
Dry tap aspirate26.66
Total cases30100

Abbreviation: BMA, bone marrow aspiration..


Table 3 . Distribution of cases based on bone marrow biopsy and immunohistochemistry findings..

IHC diagnosisN of cases%
CLL1240
CLL1240
Non-CLL1860
NHL (unclassified)930
Mantle cell lymphoma413.3
Marginal zone lymphoma00
Hairy cell leukemia00
Follicular lymphoma310
Diffuse large B cell lymphoma26.66
Total cases30100

Abbreviations: CLL, chronic lymphocytic leukemia; IHC, immuno-histochemistry; NHL, non-Hodgkin lymphoma..


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