VR09 cell line: an EBV-positive lymphoblastoid cell line with in vivo characteristics of diffuse large B cell lymphoma of activated B-cell type

PLoS One. 2012;7(12):e52811. doi: 10.1371/journal.pone.0052811. Epub 2012 Dec 21.

Abstract

Background: small B-cell neoplasms can show plasmacytic differentiation and may potentially progress to aggressive lymphoma (DLBCL). Epstein-Barr virus (EBV) infection may cause the transformation of malignant cells in vitro.

Design and method: we established VR09 cell line with plasmacytic differentiation, obtained from a case of atypical, non-CLL B-cell chronic lymphoproliferative disease with plasmacytic features. We used flow cytometry, immunohistochemistry, polymerase chain reaction, cytogenetic analysis and florescence in situ hybridization in the attempt at thoroughly characterizing the cell line. We showed VR09 tumorigenic potential in vivo, leading to the development of activated DLBCL with plasmacytic features.

Results: VR09 cells displayed plasmacytic appearance and grew as spherical tumors when inoculated subcutaneously into immunodeficient Rag2(-/-) γ-chain(-/-) mice. VR09 cell line and tumors displayed the phenotype of activated stage of B cell maturation, with secretory differentiation (CD19+ CD20+ CD79a+ CD79b+/- CD138+ cyclin D1- Ki67 80% IgM+ IgD+ MUM1+ MNDA+ CD10- CD22+ CD23+ CD43+ K+, λ- Bcl2+ Bcl6-) and they presented episomal EBV genome, chromosome 12 trisomy, lack of c-MYC rearrangement and Myd88 gene mutation, presence of somatic hypermutation in the VH region, and wild-type p53.

Conclusion: This new EBV-positive cell line may be useful to further characterize in vivo activated DLBCL with plasmacytic features.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Animals
  • CARD Signaling Adaptor Proteins / genetics
  • CD79 Antigens / genetics
  • Cell Cycle
  • Cell Line, Tumor
  • Disease Models, Animal
  • Guanylate Cyclase / genetics
  • Herpesvirus 4, Human / genetics
  • Humans
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Variable Region / genetics
  • Immunophenotyping
  • In Situ Hybridization, Fluorescence
  • Karyotype
  • Lymphoma, Large B-Cell, Diffuse / genetics*
  • Lymphoma, Large B-Cell, Diffuse / metabolism*
  • Lymphoma, Large B-Cell, Diffuse / pathology
  • Lymphoma, Large B-Cell, Diffuse / virology
  • Male
  • Mice
  • Mice, Knockout
  • Mutation
  • Myeloid Differentiation Factor 88 / genetics
  • Transplantation, Heterologous
  • Tumor Suppressor Protein p53 / genetics

Substances

  • CARD Signaling Adaptor Proteins
  • CD79 Antigens
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Variable Region
  • Myeloid Differentiation Factor 88
  • Tumor Suppressor Protein p53
  • CARD11 protein, human
  • Guanylate Cyclase

Grant support

This work was supported by Italian Ministry of University and Scientific Research (PRIN 2005, PRIN 2007), Fondazione CARIVERONA, (Bando 2003 and 2008) and Fondazione AIRC (MFAG 2008). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.