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Journal of Virology, December 1999, p. 10525-10530, Vol. 73, No. 12
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

An Epstein-Barr Virus That Expresses Only the First 231 LMP1 Amino Acids Efficiently Initiates Primary B-Lymphocyte Growth Transformation

Kenneth M. Kaye,1,* Kenneth M. Izumi,2 Hong Li,1 Eric Johannsen,2 David Davidson,2,dagger Richard Longnecker,2,Dagger and Elliott Kieff2

Department of Medicine1 and Departments of Microbiology and Molecular Genetics,2 Harvard Medical School, Channing Laboratory, Brigham and Women's Hospital, Boston, Massachusetts 02115

Received 24 May 1999/Accepted 2 September 1999

An Epstein-Barr virus (EBV) recombinant (MS231) that expresses the first 231 amino acids (aa) of LMP1 and is truncated 155 aa before the carboxyl terminus transformed resting B lymphocytes into lymphoblastoid cell lines (LCLs) only when the infected cells were grown on fibroblast feeder cells (K. M. Kaye et al., J. Virol. 69:675-683, 1995). Higher-titer MS231 virus has now been compared to wild-type (WT) EBV recombinants for the ability to cause resting primary B-lymphocyte transformation. Unexpectedly, MS231 is as potent as WT EBV recombinants in causing infected B lymphocytes to proliferate in culture for up to 5 weeks. When more than one transforming event is initiated in a microwell, the MS231 recombinant supports efficient long-term LCL outgrowth and fibroblast feeder cells are not required. However, with limited virus input, MS231-infected cells differed in their growth from WT virus-infected cells as early as 6 weeks after infection. In contrast to WT virus-infected cells, most MS231-infected cells could not be grown into long-term LCLs. Thus, the LMP1 amino-terminal 231 aa are sufficient for initial growth transformation but the carboxyl-terminal 155 aa are necessary for efficient long-term outgrowth. Despite the absence of the carboxyl-terminal 155 aa, MS231- and WT-transformed LCLs are similar in latent EBV gene expression, in ICAM-1 and CD23 expression, and in NF-kappa B and c-jun N-terminal kinase activation. MS231 recombinant-infected LCLs, however, require 16- to 64-fold higher cell density than WT-infected LCLs for regrowth after limiting dilution. These data indicate that the LMP1 carboxyl-terminal 155 aa are important for growth at lower cell density and appear to reduce dependence on paracrine growth factors.

* Corresponding author. Mailing address: Channing Laboratory, Brigham and Women's Hospital, 181 Longwood Ave., Boston, MA 02115. Phone: (617) 525-4256. Fax: (617) 525-4251. E-mail: kkaye{at}rics.bwh.harvard.edu.

dagger Present address: GelTex Pharmaceuticals, Inc., Waltham, MA 02154.

Dagger Present address: Department of Microbiology and Immunology, Northwestern University, Chicago, Ill.

Journal of Virology, December 1999, p. 10525-10530, Vol. 73, No. 12
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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