Murine Dendritic Cells for Immunotherapy and Vaccine Development: Generation, Optimization and Transduction
Introduction: Dendritic cells (DCs) play crucial roles in cellular immunity as the most powerful antigen presenting cells. They have been widely used for antigen delivery in vivo and in vitro. There are different ways to generate DCs and also gene transduction. In this study we introduce some optimi...
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| Format: | Article |
| Language: | English |
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Pasteur Institute of Iran
2020-12-01
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| Series: | Vaccine Research |
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| Online Access: | http://vacres.pasteur.ac.ir/article-1-226-en.pdf |
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| author | Mona Sadat Larijani Maryam Mashhadi Abolghasem Shirazi Amitis Ramezani Azam Bolhassani Mohammad Hassan Pouriayevali Seyed Mehdi Sadat |
| author_facet | Mona Sadat Larijani Maryam Mashhadi Abolghasem Shirazi Amitis Ramezani Azam Bolhassani Mohammad Hassan Pouriayevali Seyed Mehdi Sadat |
| author_sort | Mona Sadat Larijani |
| collection | DOAJ |
| description | Introduction: Dendritic cells (DCs) play crucial roles in cellular immunity as the most powerful antigen presenting cells. They have been widely used for antigen delivery in vivo and in vitro. There are different ways to generate DCs and also gene transduction. In this study we introduce some optimization in order to produce high amount of well differentiated murine DCs for potential immunotherapy and vaccine development applications. Methods: Murine bone marrow cells were isolated from male BALB/c mice and the cells were cultured with complete RPMI in presence of the same ratio of IL-4 and GM-CSF. Some changes were made in the medium and the lysis buffer applications to increase the differentiation rate of the cells. Lentiviral virions were applied to transfer the genes of interest to DCs with no pre-maturation steps. CD11c, MHC-II, CD80 and CD86 were assessed by flow cytometry. Results: The optimized steps led to significant increase in number of the isolated cells. IL-4 usage in a similar dose to GM-CSF led to macrophage formation inhibition. Lentiviral virions resulted in successful gene delivery along with well-maturated DCs. Conclusion: The introduced optimized steps could be followed in different DC applications by using lentiviral virions to transduce DCs, independent of the pre-maturation steps. |
| first_indexed | 2024-04-13T16:31:46Z |
| format | Article |
| id | doaj.art-d05b62d72460475e9653bc908541b22c |
| institution | Directory Open Access Journal |
| issn | 2383-2819 2423-4923 |
| language | English |
| last_indexed | 2024-04-13T16:31:46Z |
| publishDate | 2020-12-01 |
| publisher | Pasteur Institute of Iran |
| record_format | Article |
| series | Vaccine Research |
| spelling | doaj.art-d05b62d72460475e9653bc908541b22c2022-12-22T02:39:34ZengPasteur Institute of IranVaccine Research2383-28192423-49232020-12-01722227http://dx.doi.org/10.52547/vacres.7.2.22Murine Dendritic Cells for Immunotherapy and Vaccine Development: Generation, Optimization and TransductionMona Sadat Larijani0https://orcid.org/0000-0003-3681-5319Maryam Mashhadi Abolghasem Shirazi1https://orcid.org/0000-0001-6639-7221Amitis Ramezani 2https://orcid.org/0000-0001-7233-5828Azam Bolhassani3https://orcid.org/0000-0001-7363-7406Mohammad Hassan Pouriayevali4https://orcid.org/0000-0003-0133-1517Seyed Mehdi Sadat5https://orcid.org/0000-0001-7739-179XHepatitis, AIDS and Blood borne diseases Department, Pasteur Institute of Iran, Tehran, IranMolecular Virology Department, Pasteur Institute of Iran, Tehran, IranClinical Research Department, Pasteur Institute of Iran, Tehran, IranHepatitis, AIDS and Blood borne diseases Department, Pasteur Institute of Iran, Tehran, IranDepartment of Arboviruses and Viral Hemorrhagic Fevers (National Reference Laboratory), Pasteur Institute of Iran, Tehran, IranHepatitis, AIDS and Blood borne diseases Department, Pasteur Institute of Iran, Tehran, IranIntroduction: Dendritic cells (DCs) play crucial roles in cellular immunity as the most powerful antigen presenting cells. They have been widely used for antigen delivery in vivo and in vitro. There are different ways to generate DCs and also gene transduction. In this study we introduce some optimization in order to produce high amount of well differentiated murine DCs for potential immunotherapy and vaccine development applications. Methods: Murine bone marrow cells were isolated from male BALB/c mice and the cells were cultured with complete RPMI in presence of the same ratio of IL-4 and GM-CSF. Some changes were made in the medium and the lysis buffer applications to increase the differentiation rate of the cells. Lentiviral virions were applied to transfer the genes of interest to DCs with no pre-maturation steps. CD11c, MHC-II, CD80 and CD86 were assessed by flow cytometry. Results: The optimized steps led to significant increase in number of the isolated cells. IL-4 usage in a similar dose to GM-CSF led to macrophage formation inhibition. Lentiviral virions resulted in successful gene delivery along with well-maturated DCs. Conclusion: The introduced optimized steps could be followed in different DC applications by using lentiviral virions to transduce DCs, independent of the pre-maturation steps.http://vacres.pasteur.ac.ir/article-1-226-en.pdfdendritic cellsmurineoptimizationtransductionlentiviral vectors |
| spellingShingle | Mona Sadat Larijani Maryam Mashhadi Abolghasem Shirazi Amitis Ramezani Azam Bolhassani Mohammad Hassan Pouriayevali Seyed Mehdi Sadat Murine Dendritic Cells for Immunotherapy and Vaccine Development: Generation, Optimization and Transduction Vaccine Research dendritic cells murine optimization transduction lentiviral vectors |
| title | Murine Dendritic Cells for Immunotherapy and Vaccine Development: Generation, Optimization and Transduction |
| title_full | Murine Dendritic Cells for Immunotherapy and Vaccine Development: Generation, Optimization and Transduction |
| title_fullStr | Murine Dendritic Cells for Immunotherapy and Vaccine Development: Generation, Optimization and Transduction |
| title_full_unstemmed | Murine Dendritic Cells for Immunotherapy and Vaccine Development: Generation, Optimization and Transduction |
| title_short | Murine Dendritic Cells for Immunotherapy and Vaccine Development: Generation, Optimization and Transduction |
| title_sort | murine dendritic cells for immunotherapy and vaccine development generation optimization and transduction |
| topic | dendritic cells murine optimization transduction lentiviral vectors |
| url | http://vacres.pasteur.ac.ir/article-1-226-en.pdf |
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