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VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma
Published on: December 28, 2015
Improved clonality detection in Hodgkin lymphoma using the BIOMED-2-based heavy and kappa chain assay: a
Gustavo Tapia1, Carolina Sanz, José L Mate
1Department of Pathology, Hospital Germans Trias i Pujol, Badalona, Catalonia, Spain. gustavotapiam@hotmail.com
Insights
The BIOMED-2 polymerase chain reaction (PCR) assay can detect clonality in up to 25% of Hodgkin lymphoma (HL) cases using formalin-fixed, paraffin-embedded (FFPE) tissue. This method improves clonality detection in HL, even without microdissection.
Area of Science:
- Hematology
- Oncology
- Molecular Biology
Background:
- BIOMED-2 polymerase chain reaction (PCR) protocols are established for detecting clonality in non-Hodgkin B cell lymphomas.
- Validation of these protocols for Hodgkin lymphoma (HL) using formalin-fixed, paraffin-embedded (FFPE) tissue is limited.
- Assessing the sensitivity of BIOMED-2 protocols in non-microdissected HL samples is crucial for diagnostic accuracy.
Purpose of the Study:
- To evaluate the sensitivity of the BIOMED-2 PCR protocol for clonality detection in HL.
- To assess the utility of BIOMED-2 protocols on non-microdissected, FFPE tissue from HL cases.
- To determine the clonality detection rate in classic HL (cHL) and nodular lymphocyte-predominant HL (NLPHL).
Main Methods:
- Studied 69 consecutive HL cases (61 cHL, 8 NLPHL).
- Assessed IGH and IGK clonality using BIOMED-2 protocols on FFPE tissue.
- Evaluated CD30-positive cell counts, CD20-positive cell density, and tumor cell immunophenotype.
Main Results:
- Clonality was detected in 15 out of 58 assessable cHL cases (25.9%).
- IGH clonality was found in 15.5% and IGK clonality in 20.7% of cHL cases.
- No clonal rearrangements were detected in the eight NLPHL cases.
Conclusions:
- Combined IGH and IGK rearrangement analysis via BIOMED-2 protocols enhances clonality detection in HL.
- The BIOMED-2 protocol demonstrates utility for clonality detection in non-microdissected FFPE HL tissue.
- Clonality detection rates in cHL showed a trend towards improvement with increased Reed-Sternberg cell density and decreased background B cells.
Aims:
Although BIOMED-2 polymerase chain reaction (PCR) standardization protocols allow clonality detection in nearly 100% of non-Hodgkin B cell lymphomas, they have not been widely validated for Hodgkin lymphoma (HL). Our aim was to assess BIOMED-2 protocol sensitivity when using non-microdissected, formalin-fixed, paraffin-embedded (FFPE) tissue from HL cases.
Methods And Results:
We studied 69 consecutive HL cases, of which 61 corresponded to classic HL (cHL) and eight to nodular lymphocyte-predominant HL (NLPHL). CD30-positive cell numbers (<10, 10-25 or >25 per ×200 field), background CD20-positive cell density (low or high) and tumour cell immunophenotype were evaluated. IGH and IGK clonality was assessed on FFPE tissue following BIOMED-2 protocols. Of the 58 assessable cHL cases, 15 (25.9%) exhibited IGH and/or IGK clonality; IGH clonality was shown by nine (15.5%) and IGK clonality by 12 (20.7%). Clonality detection rates in cHL improved as CD30-positive Reed-Sternberg (RS) cell density increased and CD20-positive B cell density decreased, although these correlations did not reach statistical significance. Of the eight NLPHL cases studied, none showed clonal rearrangement.
Conclusions:
Combined study of IGH and IGK rearrangement according to BIOMED-2 protocols improves clonality detection rate (up to 25% of cases) in HL, even when working on non-microdissected FFPE tissue.
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