WFRF:(Sinclair A.)
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- Laurie, K. L. (author)
Cell-specific and efficient expression in mouse and human B cells by a novel hybrid immunoglobulin promoter in a lentiviral vector
- Article/chapterEnglish2007
Publisher, publication year, extent ...
- 2007-09-13
- Springer Science and Business Media LLC,2007
Numbers
- LIBRIS-ID:oai:lup.lub.lu.se:66f4acac-5b09-44a6-ab6f-ee23b64fd185
- https://lup.lub.lu.se/record/968888URI
- https://doi.org/10.1038/sj.gt.3303021DOI
Supplementary language notes
- Language:English
- Summary in:English
Part of subdatabase
- SwePubSwePub
Classification
Notes
- 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.
Subject headings and genre
- MEDICIN OCH HÄLSOVETENSKAP Medicinska och farmaceutiska grundvetenskaper Immunologi inom det medicinska området hsv//swe
- MEDICAL AND HEALTH SCIENCES Basic Medicine Immunology in the medical area hsv//eng
- EGFP
- gene transfer
- immunoglobulin promoter
- B cells
Added entries (persons, corporate bodies, meetings, titles ...)
- Blundell, M. P. (author)
- Baxendale, H. E. (author)
- Howe, S. J. (author)
- Sinclair, J. (author)
- Qasim, W. (author)
- Brunsberg, U (author)
- Thrasher, A. J. (author)
- Holmdahl, RikardLund University,Lunds universitet,Immunologi,Forskargrupper vid Lunds universitet,Immunology,Lund University Research Groups(Swepub:lu)infl-rho (author)
- Gustafsson, K (author)
- ImmunologiForskargrupper vid Lunds universitet (creator_code:org_t)
Related titles
- In:Gene Therapy: Springer Science and Business Media LLC14:23, s. 1623-16310969-71281476-5462
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- Brunsberg, U
- Thrasher, A. J.
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- Gustafsson, K
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- MEDICAL AND HEALTH SCIENCES
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- Gene Therapy
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- Lund University
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