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

被引：0
作者：
K L Laurie
M P Blundell
H E Baxendale
S J Howe
J Sinclair
W Qasim
U Brunsberg
A J Thrasher
R Holmdahl
K Gustafsson
机构：
[1] Molecular Immunology Unit,Department of Experimental Medical Science
[2] Wolfson Centre for Gene Therapy of Childhood Disease,undefined
[3] UCL Institute of Child Health,undefined
[4] Infectious Diseases and Microbiology Unit,undefined
[5] UCL Institute of Child Health,undefined
[6] Section for Medical Inflammation Research,undefined
[7] Lund University,undefined
[8] Great Ormond Street Hospital for Children NHS Trust,undefined
来源：
Gene Therapy | 2007年 / 14卷
关键词：
immunoglobulin promoter; B cells; gene transfer; EGFP;
D O I：
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学科分类号：
摘要：
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 3′ 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.
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页码：1623 / 1631
页数：8
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