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Journal of Virology, July 2003, p. 7341-7351, Vol. 77, No. 13
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.13.7341-7351.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Lentivirus Vectors Incorporating the Immunoglobulin Heavy Chain Enhancer and Matrix Attachment Regions Provide Position-Independent Expression in B Lymphocytes

Carolyn Lutzko,1,2* Dinithi Senadheera,1 Dianne Skelton,1 Denise Petersen,1 and Donald B. Kohn1,2

Division of Research Immunology and Bone Marrow Transplantation at Childrens Hospital Los Angeles,1 Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, California2

Received 21 November 2002/ Accepted 2 April 2003

In the present studies we developed lentivirus vectors with regulated, consistent transgene expression in B lymphocytes by incorporating the immunoglobulin heavy chain enhancer (Eµ) with and without associated matrix attachment regions (EµMAR) into lentivirus vectors. Incorporation of these fragments upstream of phosphoglycerate kinase (PGK) or cytomegalovirus promoters resulted in a two- to threefold increase in enhanced green fluorescent protein (EGFP) mean fluorescence intensity (MFI) in B-lymphoid but not T-lymphoid, myeloid, fibroblast, or carcinoma cell lines. A 1-log increase in EGFP expression was observed in B-lymphoid cells (but not myeloid cells) differentiated from human CD34+ progenitors in vitro transduced with Eµ- and EµMAR-containing lentivectors. Lastly, we evaluated the expression from the EµMAR element in mice 2 to 24 weeks posttransplant with transduced hematopoietic stem cells. In mice receiving vectors with the Eµ and EµMAR elements upstream of the PGK promoter, there was a 2- to 10-fold increase in EGFP expression in B cells (but not other cell types). Evaluation of the coefficient of variation of expression among different cell types demonstrated that consistent, position-independent transgene expression was observed exclusively in B cells transduced with the EµMAR-containing vector and not other cells types or vectors. Proviral genomes with the EµMAR element had increased chromatin accessibility, which likely contributed to the position independence of expression in B lymphocytes. In summary, incorporation of the EµMAR element in lentivirus vectors resulted in enhanced, position-independent expression in primary B lymphocytes. These vectors provide a useful tool for the study of B-lymphocyte biology and the development of gene therapy for disorders affecting B lymphocytes, such as immune deficiencies.

* Corresponding author. Mailing address: Childrens Hospital Los Angeles, 4650 Sunset Blvd. MS #62, Los Angeles, CA 90027. Phone: (323) 669-4125. Fax: (323) 667-1021. E-mail: clutzko{at}chla.usc.edu.

Journal of Virology, July 2003, p. 7341-7351, Vol. 77, No. 13
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.13.7341-7351.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Hendrickson, B., Senadheera, D., Mishra, S., Bui, K. C. T., Wang, X., Chan, B., Petersen, D., Pepper, K., Lutzko, C. (2007). Development of Lentiviral Vectors with Regulated Respiratory Epithelial Expression In Vivo. Am. J. Respir. Cell Mol. Bio. 37: 414-423 [Abstract] [Full Text]  
  • Yang, L., Bailey, L., Baltimore, D., Wang, P. (2006). Targeting lentiviral vectors to specific cell types in vivo. Proc. Natl. Acad. Sci. USA 103: 11479-11484 [Abstract] [Full Text]  


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