In vivo generation of CAR T cells in the presence of human myeloid cells
Pre-clinical humanized mouse models are a powerful tool to evaluate immunotherapies. NSG-SGM3 mice reconstituted with human stem cells (huSGM3) develop pronounced human myeloid cells due to transgenic expression of stem cell factor, granulocyte-macrophage colony-stimulating factor, and interleukin-3...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-09-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050122000821 |
| _version_ | 1828834363944468480 |
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| author | Naphang Ho Shiwani Agarwal Michela Milani Alessio Cantore Christian J. Buchholz Frederic B. Thalheimer |
| author_facet | Naphang Ho Shiwani Agarwal Michela Milani Alessio Cantore Christian J. Buchholz Frederic B. Thalheimer |
| author_sort | Naphang Ho |
| collection | DOAJ |
| description | Pre-clinical humanized mouse models are a powerful tool to evaluate immunotherapies. NSG-SGM3 mice reconstituted with human stem cells (huSGM3) develop pronounced human myeloid cells due to transgenic expression of stem cell factor, granulocyte-macrophage colony-stimulating factor, and interleukin-3 (IL-3) compared with the widely used humanized NSG (huNSG) model. We assessed in vivo generation of CD19-CAR T cells in huSGM3 mice upon single intravenous injection of the T cell-specific lentiviral vectors (LVs) CD4-LV and CD8-LV. While in vivo CAR T cell generation was clearly detectable in individual mice, generation appeared less efficient than previously observed for huNSG mice. Especially for the CD4-LV group, this correlated with increased IL-15 and decreased GM-CSF levels, indicating activation of monocytes and macrophages. Co-culture assays identified macrophages as a potential barrier for gene transfer. Refining CD4-LV and CD8-LV with a less immunogenic surface by using modified packaging cells substantially improved the transduction of lymphocytes in vitro in the presence of macrophages, as well in vivo in huSGM3 mice. Notably, two mice that developed less CAR T cells showed high interferon-α or -β levels before vector injection. Our data emphasize the relevance of innate immune responses for in vivo generation of CAR T cells, which can be overcome by vector surface engineering. |
| first_indexed | 2024-12-12T17:38:00Z |
| format | Article |
| id | doaj.art-e78a0da4c10e4bfabac0ead54b2a2f99 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-12-12T17:38:00Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-e78a0da4c10e4bfabac0ead54b2a2f992022-12-22T00:17:09ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012022-09-0126144156In vivo generation of CAR T cells in the presence of human myeloid cellsNaphang Ho0Shiwani Agarwal1Michela Milani2Alessio Cantore3Christian J. Buchholz4Frederic B. Thalheimer5Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Paul-Ehrlich-Straße 51-59, 63225 Langen, Germany; Frankfurt Cancer Institute, Goethe University, 60590 Frankfurt am Main, GermanyMolecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Paul-Ehrlich-Straße 51-59, 63225 Langen, GermanySan Raffaele Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific Institute, 20132 Milan, ItalySan Raffaele Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy; Vita-Salute San Raffaele University, 20132 Milan, ItalyMolecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Paul-Ehrlich-Straße 51-59, 63225 Langen, Germany; Frankfurt Cancer Institute, Goethe University, 60590 Frankfurt am Main, Germany; Division of Medical Biotechnology, Paul-Ehrlich-Institut, 63225 Langen, GermanyMolecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Paul-Ehrlich-Straße 51-59, 63225 Langen, Germany; Frankfurt Cancer Institute, Goethe University, 60590 Frankfurt am Main, Germany; Corresponding author Frederic B. Thalheimer, Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Paul-Ehrlich-Straße 51-59, 63225 Langen, Germany.Pre-clinical humanized mouse models are a powerful tool to evaluate immunotherapies. NSG-SGM3 mice reconstituted with human stem cells (huSGM3) develop pronounced human myeloid cells due to transgenic expression of stem cell factor, granulocyte-macrophage colony-stimulating factor, and interleukin-3 (IL-3) compared with the widely used humanized NSG (huNSG) model. We assessed in vivo generation of CD19-CAR T cells in huSGM3 mice upon single intravenous injection of the T cell-specific lentiviral vectors (LVs) CD4-LV and CD8-LV. While in vivo CAR T cell generation was clearly detectable in individual mice, generation appeared less efficient than previously observed for huNSG mice. Especially for the CD4-LV group, this correlated with increased IL-15 and decreased GM-CSF levels, indicating activation of monocytes and macrophages. Co-culture assays identified macrophages as a potential barrier for gene transfer. Refining CD4-LV and CD8-LV with a less immunogenic surface by using modified packaging cells substantially improved the transduction of lymphocytes in vitro in the presence of macrophages, as well in vivo in huSGM3 mice. Notably, two mice that developed less CAR T cells showed high interferon-α or -β levels before vector injection. Our data emphasize the relevance of innate immune responses for in vivo generation of CAR T cells, which can be overcome by vector surface engineering.http://www.sciencedirect.com/science/article/pii/S2329050122000821macrophagein vivo gene deliveryshieldingT cell targeted lentiviral vectorshumanized mouse modelinnate immune response |
| spellingShingle | Naphang Ho Shiwani Agarwal Michela Milani Alessio Cantore Christian J. Buchholz Frederic B. Thalheimer In vivo generation of CAR T cells in the presence of human myeloid cells Molecular Therapy: Methods & Clinical Development macrophage in vivo gene delivery shielding T cell targeted lentiviral vectors humanized mouse model innate immune response |
| title | In vivo generation of CAR T cells in the presence of human myeloid cells |
| title_full | In vivo generation of CAR T cells in the presence of human myeloid cells |
| title_fullStr | In vivo generation of CAR T cells in the presence of human myeloid cells |
| title_full_unstemmed | In vivo generation of CAR T cells in the presence of human myeloid cells |
| title_short | In vivo generation of CAR T cells in the presence of human myeloid cells |
| title_sort | in vivo generation of car t cells in the presence of human myeloid cells |
| topic | macrophage in vivo gene delivery shielding T cell targeted lentiviral vectors humanized mouse model innate immune response |
| url | http://www.sciencedirect.com/science/article/pii/S2329050122000821 |
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