Related Experiment Video
Updated: Oct 10, 2026
Isolation and Quantification of Epstein-Barr Virus from the P3HR1 Cell Line
Published on: September 28, 2022
The characteristics of Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma: comparison between EBV(+) and
T Kuze1, N Nakamura, Y Hashimoto
1First Department of Pathology, Fukushima Medical University, Fukushima 960-1295, Japan. kuze.tetsuo@toaeiyo.ne.jp
Insights
Epstein-Barr virus (EBV) infection in diffuse large B-cell lymphoma (DLBCL) is linked to non-functional immunoglobulin heavy chain genes. This EBV(+) DLBCL characteristic suggests a distinct mutational profile compared to EBV(-) DLBCL.
Area of Science:
- Oncology
- Virology
- Immunology
Background:
- Diffuse large B-cell lymphoma (DLBCL) is an aggressive non-Hodgkin lymphoma.
- Epstein-Barr virus (EBV) coinfection is observed in a subset of DLBCL cases.
- The specific characteristics and implications of EBV infection in DLBCL remain incompletely understood.
Purpose of the Study:
- To investigate the clinicopathological and molecular features of DLBCL associated with EBV infection.
- To compare the characteristics of EBV-positive (EBV(+)) DLBCL with EBV-negative (EBV(-)) DLBCL.
- To analyze the immunoglobulin heavy chain (IgH) gene mutation status in EBV(+) and EBV(-) DLBCL.
Main Methods:
- Analysis of 114 DLBCL cases using RNA in situ hybridization for EBV-encoded RNA 1 (EBER1).
- Immunohistochemical detection of EBV latent membrane protein 1 (LMP1), EBV nuclear antigen 2 (EBNA2), CD30, Bcl-6, and immunoglobulin (Ig).
- Sequencing of rearranged Ig heavy chain (IgH) variable (V) region genes to assess somatic mutation frequency and types.
Main Results:
- EBV infection (EBER1 positivity) was detected in 11.4% of DLBCL cases.
- EBV(+) DLBCL cases showed distinct expression profiles for CD30, Bcl-6, and Ig compared to EBV(-) DLBCL.
- A significant proportion (40%) of EBV(+) DLBCL cases exhibited crippling mutations in the IgH gene, leading to non-functional genes, unlike EBV(-) DLBCL (0%).
Conclusions:
- Both EBV(+) and EBV(-) DLBCL appear to originate from germinal center (GC) or post-GC B cells.
- EBV(+) DLBCL is frequently characterized by a non-functional IgH gene due to crippling mutations.
- These findings highlight a distinct molecular pathway in EBV(+) DLBCL, potentially impacting B-cell receptor function.
Abstract:
We have investigated 114 cases with diffuse large B-cell lymphoma (DLBCL) to clarify the characteristics of DLBCL with Epstein-Barr virus (EBV) infection. Thirteen cases (11.4%) showed EBV-encoded RNA 1 (EBER1) signals by RNA in situ hybridization. EBV-encoded latent membrane protein 1 (LMP1) and EBV-encoded nuclear antigen 2 (EBNA2) were expressed in 11 and 4 cases, respectively. Expression of CD30, Bcl-6 and immunoglobulin (Ig) was found in 92%, 31% and 23% with EBV(+) DLBCL, and in 15%, 79% and 82% with EBV(-) DLBCL, respectively. The sequence of rearranged Ig heavy chain (IgH) variable (V) region gene was analyzed in 5 cases with EBV(+) DLBCL and 61 cases with EBV(-) DLBCL. Somatic mutation was found in all cases except one with EBV(-) DLBCL. Average mutation frequency was 9.6% in EBV(+) DLBCL vs. 11.5% in EBV(-) DLBCL. The rates of replacement mutation vs. silent mutation (R / S values) in complementarity determining region II and framework region III were 2.7 and 1.5 in EBV(+) DLBCL, 2.6 and 1.4 in EBV(-) DLBCL. Crippling mutation generating a stop codon was found in 2 of 5 cases (40%) with EBV(+) DLBCL, but none of 61 cases (0%) with EBV(-) DLBCL. These findings suggest that EBV(+) DLBCL and EBV(-) DLBCL were both derived from germinal center (GC) or post-GC B cells, and EBV(+) DLBCL frequently have a non-functional IgH gene owing to crippling mutation.
