Efficient lentiviral transduction of Herpesvirus saimiri immortalized T cells as a model for gene therapy in primary immunodeficiencies

Toscano, Miguel G.; Frecha, Cecilia; Ortega, Consuelo; Santamarı́a, Manuel; Martin, F. Elizabeth; Molina, Ignacio J.

doi:10.1038/sj.gt.3302259

Cited by 28 publications

(17 citation statements)

References 37 publications

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“…26 Very recently, we have demonstrated that HVSimmortalized T cells from WAS patients can efficiently be transduced by lentiviral vectors and are therefore a useful model for WAS gene therapy. 27 In this study, we show that a 500 bp fragment from the proximal WASP gene promoter, in the context of a SIN lentiviral vector, achieves hematopoietic-specific and physiologically relevant levels of WASp.…”

Section: Introductionmentioning

confidence: 67%

Lentiviral vectors transcriptionally targeted to hematopoietic cells by WASP gene proximal promoter sequences

et al. 2005

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Section: Introductionmentioning

confidence: 67%

Lentiviral vectors transcriptionally targeted to hematopoietic cells by WASP gene proximal promoter sequences

et al. 2005

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“…The packaging and production of the lentiviral vector were carried out as previously described (17). Briefly, the lentiviral shuttle plasmid and auxiliary packaging plasmid were constructed.…”

Section: Methodsmentioning

confidence: 99%

Islet-1 promotes the cardiac-specific differentiation of mesenchymal stem cells through the regulation of histone acetylation

Yin

Lin

et al. 2014

International Journal of Molecular Medicine

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The aim of the present study was to investigate the effects of Islet-1 on the process of mesenchymal stem cell (MSC) differentiation into cardiomyocyte-like cells and to elucidate the possible mechanisms involved. Lentiviral vectors expressing Islet-1 (Lenti-Islet-1) were constructed and used for C3H10T1/2 cell transfection. Cell morphology was observed. Cardiac-related genes and proteins were detected by qPCR and western blot analysis. Epigallocatechin gallate (EGCG) was used as an inhibitor of acetylated histone H3 (AcH3). AcH3 was detected by chromatin immunoprecipitation. Cells overexpressing Islet-1 tended to change into fibroblast-like cells and were arranged in the same direction. The enhanced expression of GATA binding protein 4 (Gata4), NK2 homeobox 5 (Nkx2.5), myocyte enhancer factor 2C (Mef2c) and cardiac troponin T (cTnT) was observed in the cells overexpressing Islet-1 following transfection with Lenti-Islet-1. However, the expression of hepatocyte-, bone- and neuronal-specific markers was not affected by Islet-1. The AcH3 relative amount increased following transfection with Lenti-Islet-1, which was associated with the enhanced expression of Gata4, Nkx2.5 and Mef2c in these cells. The expression of Gata4, Nkx2.5 and Mef2c in the C3H10T1/2 cells transfected with Lenti-Islet-1 and treated with EGCG was reduced following treatment with EGCG. The data presented in this study indicate that Islet-1 specifically induces the differentiation of C3H10T1/2 cells into cardiomyocyte-like cells, and one of the mechanisms involved is the regulation of histone acetylation.

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“…Briefly, (1) Vector plasmid (SE, WE or AWE), (2) pCMVDR8.9 and (3) Envelope plasmid pMD.G as described previously. 44 Viral titres (transduction units per ml) were calculated by FACS based on the initial amount of target cells (Jurkat cell line) and the percentage of eGFP + cells detected in the linear range of serial dilutions of the supernatant, ranging 5 Â 10 6 -1 Â 10 7 transducing unit (TU) per ml in all the vector batches.…”

Section: Plasmids Lentiviral Constructs and Vector Productionmentioning

confidence: 99%

Improved lentiviral vectors for Wiskott–Aldrich syndrome gene therapy mimic endogenous expression profiles throughout haematopoiesis

et al. 2008

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Wiskott-Aldrich syndrome (WAS) gene therapy requires highly efficient and well-controlled vectors. Here we studied the performance of a lentiviral vector (LV) harbouring a 500-bp fragment of the WAS proximal promoter (WW), which we previously characterized as haematopoietic-specific and capable of restoring WAS phenotype in patients' T cells. We used an LV (WE) expressing eGFP to evaluate whether this promoter was following the expression pattern of endogenous WASp. Transgene expression was analysed in WE-transduced hCD34 + population and its progeny after in vitro and in vivo differentiation in the Rag 2 À/À , gc À/À humanized mouse. We revealed very poor expression from the WE internal promoter in macrophages and erythroid cells. Therefore, we designed a novel LV including a fragment of the alternative WAS promoter in WE vector (AWE). This new vector sustained high transgene levels along the whole lymphoid lineage in vivo. Most importantly, the performance of AWE vector was highly superior to WE vector since AWE clearly improved transgene levels in in vitro and in vivo hCD34 + -derived macrophages, erythroid cells, megakaryocytes and B cells while supporting a high expression in human T cells. This emphasizes that it is a suitable LV backbone for gene therapy of haematopoietic diseases such as WAS. Gene Therapy (2008) 15, 930-941;

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Efficient lentiviral transduction of Herpesvirus saimiri immortalized T cells as a model for gene therapy in primary immunodeficiencies

Cited by 28 publications

References 37 publications

Lentiviral vectors transcriptionally targeted to hematopoietic cells by WASP gene proximal promoter sequences

Lentiviral vectors transcriptionally targeted to hematopoietic cells by WASP gene proximal promoter sequences

Islet-1 promotes the cardiac-specific differentiation of mesenchymal stem cells through the regulation of histone acetylation

Improved lentiviral vectors for Wiskott–Aldrich syndrome gene therapy mimic endogenous expression profiles throughout haematopoiesis

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