Epstein-Barr virus lacking latent membrane protein 2 immortalizes B cells with efficiency indistinguishable from that of wild-type virus

J Gen Virol. 1999 Aug;80 ( Pt 8):2193-2203. doi: 10.1099/0022-1317-80-8-2193.

Abstract

Epstein-Barr virus (EBV) is a human herpesvirus that efficiently transforms and immortalizes human primary B lymphocytes. In this study, the role of latent membrane protein 2 (LMP2) in EBV growth transformation was investigated. LMP2 is a virally encoded membrane protein expressed in EBV-immortalized B cells previously shown to be nonessential for EBV transformation. However, a recent study reported that LMP2 may be an important determinant for efficient B cell transformation (Brielmeier et al., Journal of General Virology 77, 2807-2818, 1996). In this study a deletion mutation was introduced into the LMP2 gene using an E. coli mini-EBV construct containing sufficient EBV DNA to result in growth transformation of primary B cells. In an alternative approach, the introduction of the gene encoding the enhanced green fluorescent protein (EGFP) by homologous recombination into the LMP2 gene of EBV strain B95-8, generating the same LMP2 deletion mutation is reported. Careful quantification of B cell transformation using the EGFP+ LMP2- recombinant virus determined that in liquid culture medium or in culture medium containing soft agarose there was no difference in the ability of LMP2- virus to immortalize primary human B cells when compared to that of wild-type virus.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • B-Lymphocytes* / cytology
  • B-Lymphocytes* / virology
  • Callithrix
  • Cell Division
  • Cell Line
  • Cell Transformation, Viral*
  • Cells, Cultured
  • Cysteine Endopeptidases*
  • Gene Deletion
  • Green Fluorescent Proteins
  • Herpesvirus 4, Human / genetics
  • Herpesvirus 4, Human / physiology*
  • Humans
  • Luminescent Proteins
  • Proteins / genetics
  • Proteins / physiology*
  • Sepharose
  • Tumor Cells, Cultured

Substances

  • Luminescent Proteins
  • Proteins
  • LMP-2 protein
  • Green Fluorescent Proteins
  • Sepharose
  • Cysteine Endopeptidases