A Nontoxic Transduction Enhancer Enables Highly Efficient Lentiviral Transduction of Primary Murine T Cells and Hematopoietic Stem Cells
Lentiviral vectors have emerged as an efficient, safe therapeutic tool for gene therapy based on hematopoietic stem cells (HSCs) or T cells. However, the monitoring of transduced cells in preclinical models remains challenging because of the inefficient transduction of murine primary T cells with le...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2018-09-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050118300792 |
| _version_ | 1811280131267756032 |
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| author | Marianne Delville Tayebeh Soheili Florence Bellier Amandine Durand Adeline Denis Chantal Lagresle-Peyrou Marina Cavazzana Isabelle Andre-Schmutz Emmanuelle Six |
| author_facet | Marianne Delville Tayebeh Soheili Florence Bellier Amandine Durand Adeline Denis Chantal Lagresle-Peyrou Marina Cavazzana Isabelle Andre-Schmutz Emmanuelle Six |
| author_sort | Marianne Delville |
| collection | DOAJ |
| description | Lentiviral vectors have emerged as an efficient, safe therapeutic tool for gene therapy based on hematopoietic stem cells (HSCs) or T cells. However, the monitoring of transduced cells in preclinical models remains challenging because of the inefficient transduction of murine primary T cells with lentiviral vectors, in contrast to gammaretroviral vectors. The use of this later in preclinical proof of concept is not considered as relevant when a lentiviral vector will be used in a clinical trial. Hence, there is an urgent need to develop an efficient transduction protocol for murine cells with lentiviral vectors. Here, we describe an optimized protocol in which a nontoxic transduction enhancer (Lentiboost) enables the efficient transduction of primary murine T cells with lentiviral vectors. The optimized protocol combines low toxicity and high transduction efficiency. We achieved a high-level transduction of murine CD4+ and CD8+ T cells with a VSV-G-pseudotyped lentiviral vector with no changes in the phenotypes of transduced T cells, which were stable and long-lived in culture. This enhancer also increased the transduction of murine HSCs. Hence, use of this new transduction enhancer overcomes the limitations of lentiviral vectors in preclinical experiments and should facilitate the translation of strategies based on lentiviral vectors from the bench to the clinic. Keywords: murine CD4+ T cells, murine CD8+ T cells, murine Sca1+ cells, lentiviral transduction, Lentiboost, gene therapy |
| first_indexed | 2024-04-13T01:07:58Z |
| format | Article |
| id | doaj.art-af84fad0a9074c88ae2fe4f7bc9bca51 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-04-13T01:07:58Z |
| publishDate | 2018-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-af84fad0a9074c88ae2fe4f7bc9bca512022-12-22T03:09:16ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012018-09-0110341347A Nontoxic Transduction Enhancer Enables Highly Efficient Lentiviral Transduction of Primary Murine T Cells and Hematopoietic Stem CellsMarianne Delville0Tayebeh Soheili1Florence Bellier2Amandine Durand3Adeline Denis4Chantal Lagresle-Peyrou5Marina Cavazzana6Isabelle Andre-Schmutz7Emmanuelle Six8Laboratory of Human Lymphohematopoiesis, INSERM UMR 1163, Imagine Institute, Paris, France; Apheresis and Biotherapy Department, Necker Hospital, APHP, Paris, France; University of Paris Descartes-Sorbonne Paris Cité, Paris, France; Biotherapy Clinical Investigation Center, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpitaux de Paris, INSERM CIC 1416, Paris, FranceLaboratory of Human Lymphohematopoiesis, INSERM UMR 1163, Imagine Institute, Paris, France; Apheresis and Biotherapy Department, Necker Hospital, APHP, Paris, France; University of Paris Descartes-Sorbonne Paris Cité, Paris, France; Biotherapy Clinical Investigation Center, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpitaux de Paris, INSERM CIC 1416, Paris, FranceLaboratory of Human Lymphohematopoiesis, INSERM UMR 1163, Imagine Institute, Paris, France; University of Paris Descartes-Sorbonne Paris Cité, Paris, France; Biotherapy Clinical Investigation Center, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpitaux de Paris, INSERM CIC 1416, Paris, FranceLaboratory of Human Lymphohematopoiesis, INSERM UMR 1163, Imagine Institute, Paris, France; University of Paris Descartes-Sorbonne Paris Cité, Paris, France; Biotherapy Clinical Investigation Center, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpitaux de Paris, INSERM CIC 1416, Paris, FranceLaboratory of Human Lymphohematopoiesis, INSERM UMR 1163, Imagine Institute, Paris, France; University of Paris Descartes-Sorbonne Paris Cité, Paris, France; Biotherapy Clinical Investigation Center, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpitaux de Paris, INSERM CIC 1416, Paris, FranceLaboratory of Human Lymphohematopoiesis, INSERM UMR 1163, Imagine Institute, Paris, France; University of Paris Descartes-Sorbonne Paris Cité, Paris, France; Biotherapy Clinical Investigation Center, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpitaux de Paris, INSERM CIC 1416, Paris, FranceLaboratory of Human Lymphohematopoiesis, INSERM UMR 1163, Imagine Institute, Paris, France; Apheresis and Biotherapy Department, Necker Hospital, APHP, Paris, France; University of Paris Descartes-Sorbonne Paris Cité, Paris, France; Biotherapy Clinical Investigation Center, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpitaux de Paris, INSERM CIC 1416, Paris, FranceLaboratory of Human Lymphohematopoiesis, INSERM UMR 1163, Imagine Institute, Paris, France; University of Paris Descartes-Sorbonne Paris Cité, Paris, France; Biotherapy Clinical Investigation Center, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpitaux de Paris, INSERM CIC 1416, Paris, France; Corresponding author: Isabelle Andre-Schmutz, Laboratory of Human Lymphohematopoiesis, INSERM UMR 1163, Imagine Institute, Paris, France.Laboratory of Human Lymphohematopoiesis, INSERM UMR 1163, Imagine Institute, Paris, France; University of Paris Descartes-Sorbonne Paris Cité, Paris, France; Biotherapy Clinical Investigation Center, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpitaux de Paris, INSERM CIC 1416, Paris, FranceLentiviral vectors have emerged as an efficient, safe therapeutic tool for gene therapy based on hematopoietic stem cells (HSCs) or T cells. However, the monitoring of transduced cells in preclinical models remains challenging because of the inefficient transduction of murine primary T cells with lentiviral vectors, in contrast to gammaretroviral vectors. The use of this later in preclinical proof of concept is not considered as relevant when a lentiviral vector will be used in a clinical trial. Hence, there is an urgent need to develop an efficient transduction protocol for murine cells with lentiviral vectors. Here, we describe an optimized protocol in which a nontoxic transduction enhancer (Lentiboost) enables the efficient transduction of primary murine T cells with lentiviral vectors. The optimized protocol combines low toxicity and high transduction efficiency. We achieved a high-level transduction of murine CD4+ and CD8+ T cells with a VSV-G-pseudotyped lentiviral vector with no changes in the phenotypes of transduced T cells, which were stable and long-lived in culture. This enhancer also increased the transduction of murine HSCs. Hence, use of this new transduction enhancer overcomes the limitations of lentiviral vectors in preclinical experiments and should facilitate the translation of strategies based on lentiviral vectors from the bench to the clinic. Keywords: murine CD4+ T cells, murine CD8+ T cells, murine Sca1+ cells, lentiviral transduction, Lentiboost, gene therapyhttp://www.sciencedirect.com/science/article/pii/S2329050118300792 |
| spellingShingle | Marianne Delville Tayebeh Soheili Florence Bellier Amandine Durand Adeline Denis Chantal Lagresle-Peyrou Marina Cavazzana Isabelle Andre-Schmutz Emmanuelle Six A Nontoxic Transduction Enhancer Enables Highly Efficient Lentiviral Transduction of Primary Murine T Cells and Hematopoietic Stem Cells Molecular Therapy: Methods & Clinical Development |
| title | A Nontoxic Transduction Enhancer Enables Highly Efficient Lentiviral Transduction of Primary Murine T Cells and Hematopoietic Stem Cells |
| title_full | A Nontoxic Transduction Enhancer Enables Highly Efficient Lentiviral Transduction of Primary Murine T Cells and Hematopoietic Stem Cells |
| title_fullStr | A Nontoxic Transduction Enhancer Enables Highly Efficient Lentiviral Transduction of Primary Murine T Cells and Hematopoietic Stem Cells |
| title_full_unstemmed | A Nontoxic Transduction Enhancer Enables Highly Efficient Lentiviral Transduction of Primary Murine T Cells and Hematopoietic Stem Cells |
| title_short | A Nontoxic Transduction Enhancer Enables Highly Efficient Lentiviral Transduction of Primary Murine T Cells and Hematopoietic Stem Cells |
| title_sort | nontoxic transduction enhancer enables highly efficient lentiviral transduction of primary murine t cells and hematopoietic stem cells |
| url | http://www.sciencedirect.com/science/article/pii/S2329050118300792 |
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