Epstein-Barr virus lacking latent membrane protein 2 immortalizes B cells with efficiency indistinguishable from that of wild-type virus.
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.
Speck
Peter
P
Department of Microbiology-Immunology, Northwestern University Medical School, Room 6-231 Ward Building, 303 East Chicago Avenue, Chicago, Illinois 60611, USA1.
Kline
Kimberly A
KA
Department of Microbiology-Immunology, Northwestern University Medical School, Room 6-231 Ward Building, 303 East Chicago Avenue, Chicago, Illinois 60611, USA1.
Cheresh
Paul
P
Department of Microbiology-Immunology, Northwestern University Medical School, Room 6-231 Ward Building, 303 East Chicago Avenue, Chicago, Illinois 60611, USA1.
Longnecker
Richard
R
Department of Microbiology-Immunology, Northwestern University Medical School, Room 6-231 Ward Building, 303 East Chicago Avenue, Chicago, Illinois 60611, USA1.
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000821:virology; D002144:Callithrix; D002455:Cell Division; D002460:Cell Line; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases; D017353:Gene Deletion; D049452:Green Fluorescent Proteins; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008164:Luminescent Proteins; D011506:Proteins / Q000235:genetics / Q000502:physiology*; D012685:Sepharose; D014407:Tumor Cells, Cultured
D008164:Luminescent Proteins; D011506:Proteins; C071313:LMP-2 protein; D049452:Green Fluorescent Proteins; D012685:Sepharose; D003546:Cysteine Endopeptidases