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Cell-specific and efficient expression in mouse and human B cells by a novel hybrid immunoglobulin promoter in a lentiviral vector

Laurie, K. L.; Blundell, M. P.; Baxendale, H. E.; Howe, S. J.; Sinclair, J.; Qasim, W.; Brunsberg, U; Thrasher, A. J.; Holmdahl, Rikard LU and Gustafsson, K (2007) In Gene Therapy 14(23). p.1623-1631
Abstract
The expression of genes specifically in B cells is of great interest in both experimental immunology as well as in future clinical gene therapy. We have constructed a novel enhanced B cell-specific promoter (Igk- E) consisting of an immunoglobulin kappa (Igk) minimal promoter combined with an intronic enhancer sequence and a 30 enhancer sequence from Ig genes. The Igk- E promoter was cloned into a lentiviral vector and used to control expression of enhanced green fluorescent protein (eGFP). Transduction of murine B-cell lymphoma cell lines and activated primary splenic B cells, with IgK-E-eGFP lentivirus, resulted in expression of eGFP, as analysed by flow cytometry, whereas expression in non-B cells was absent. The specificity of the... (More)
The expression of genes specifically in B cells is of great interest in both experimental immunology as well as in future clinical gene therapy. We have constructed a novel enhanced B cell-specific promoter (Igk- E) consisting of an immunoglobulin kappa (Igk) minimal promoter combined with an intronic enhancer sequence and a 30 enhancer sequence from Ig genes. The Igk- E promoter was cloned into a lentiviral vector and used to control expression of enhanced green fluorescent protein (eGFP). Transduction of murine B-cell lymphoma cell lines and activated primary splenic B cells, with IgK-E-eGFP lentivirus, resulted in expression of eGFP, as analysed by flow cytometry, whereas expression in non-B cells was absent. The specificity of the promoter was further examined by transducing Lin bone marrow with Igk-E-eGFP lentivirus and reconstituting lethally irradiated mice. After 16 weeks flow cytometry of lymphoid tissues revealed eGFP expression by CD19(+) cells, but not by CD3(+), CD11b(+), CD11c(+) or Gr-1(+) cells. CD19(+) cells were comprised of both marginal zone B cells and recirculating follicular B cells. Activated human peripheral mononuclear cells were also transduced with Igk-E-eGFP lentivirus under conditions of selective B-cell activation. The Igk-E promoter was able to drive expression of eGFP only in CD19(+) cells, while eGFP was expressed by both spleen focus forming virus and cytomegalovirus constitutive promoters in CD19(+) and CD3(+) lymphocytes. These data demonstrate that in these conditions the Igk-E promoter is cell specific and controls efficient expression of a reporter protein in mouse and human B cells in the context of a lentiviral vector. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
EGFP, gene transfer, immunoglobulin promoter, B cells
in
Gene Therapy
volume
14
issue
23
pages
1623 - 1631
publisher
Nature Publishing Group
external identifiers
  • wos:000251252000003
  • scopus:36348946277
ISSN
0969-7128
DOI
10.1038/sj.gt.3303021
language
English
LU publication?
yes
id
66f4acac-5b09-44a6-ab6f-ee23b64fd185 (old id 968888)
date added to LUP
2008-01-29 15:16:55
date last changed
2017-07-30 03:53:16
@article{66f4acac-5b09-44a6-ab6f-ee23b64fd185,
  abstract     = {The expression of genes specifically in B cells is of great interest in both experimental immunology as well as in future clinical gene therapy. We have constructed a novel enhanced B cell-specific promoter (Igk- E) consisting of an immunoglobulin kappa (Igk) minimal promoter combined with an intronic enhancer sequence and a 30 enhancer sequence from Ig genes. The Igk- E promoter was cloned into a lentiviral vector and used to control expression of enhanced green fluorescent protein (eGFP). Transduction of murine B-cell lymphoma cell lines and activated primary splenic B cells, with IgK-E-eGFP lentivirus, resulted in expression of eGFP, as analysed by flow cytometry, whereas expression in non-B cells was absent. The specificity of the promoter was further examined by transducing Lin bone marrow with Igk-E-eGFP lentivirus and reconstituting lethally irradiated mice. After 16 weeks flow cytometry of lymphoid tissues revealed eGFP expression by CD19(+) cells, but not by CD3(+), CD11b(+), CD11c(+) or Gr-1(+) cells. CD19(+) cells were comprised of both marginal zone B cells and recirculating follicular B cells. Activated human peripheral mononuclear cells were also transduced with Igk-E-eGFP lentivirus under conditions of selective B-cell activation. The Igk-E promoter was able to drive expression of eGFP only in CD19(+) cells, while eGFP was expressed by both spleen focus forming virus and cytomegalovirus constitutive promoters in CD19(+) and CD3(+) lymphocytes. These data demonstrate that in these conditions the Igk-E promoter is cell specific and controls efficient expression of a reporter protein in mouse and human B cells in the context of a lentiviral vector.},
  author       = {Laurie, K. L. and Blundell, M. P. and Baxendale, H. E. and Howe, S. J. and Sinclair, J. and Qasim, W. and Brunsberg, U and Thrasher, A. J. and Holmdahl, Rikard and Gustafsson, K},
  issn         = {0969-7128},
  keyword      = {EGFP,gene transfer,immunoglobulin promoter,B cells},
  language     = {eng},
  number       = {23},
  pages        = {1623--1631},
  publisher    = {Nature Publishing Group},
  series       = {Gene Therapy},
  title        = {Cell-specific and efficient expression in mouse and human B cells by a novel hybrid immunoglobulin promoter in a lentiviral vector},
  url          = {http://dx.doi.org/10.1038/sj.gt.3303021},
  volume       = {14},
  year         = {2007},
}