Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway
The baculovirus Autographa californica multiple nucleopolyhedrovirus is an insect virus with a circular double-stranded DNA genome, which, among other multiple biotechnological applications, is used as an expression vector for gene delivery in mammalian cells. Nevertheless, the nonspecific immune re...
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| Format: | Artículo |
| Language: | Inglés |
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American Society for Microbiology
2021
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| Online Access: | http://hdl.handle.net/20.500.12123/10117 https://journals.asm.org/doi/10.1128/JVI.01555-20 https://doi.org/10.1128/JVI.01555-20 |
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| author | Amalfi, Sabrina Molina, Guido Nicolas Bevacqua, Romina Jimena Lopez, Maria Gabriela Taboga, Oscar Alberto Alfonso, Victoria |
| author_browse | Alfonso, Victoria Amalfi, Sabrina Bevacqua, Romina Jimena Lopez, Maria Gabriela Molina, Guido Nicolas Taboga, Oscar Alberto |
| author_facet | Amalfi, Sabrina Molina, Guido Nicolas Bevacqua, Romina Jimena Lopez, Maria Gabriela Taboga, Oscar Alberto Alfonso, Victoria |
| author_sort | Amalfi, Sabrina |
| collection | INTA Digital |
| description | The baculovirus Autographa californica multiple nucleopolyhedrovirus is an insect virus with a circular double-stranded DNA genome, which, among other multiple biotechnological applications, is used as an expression vector for gene delivery in mammalian cells. Nevertheless, the nonspecific immune response triggered by viral vectors often suppresses transgene expression. To understand the mechanisms involved in that response, in the present study, we studied the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway by using two approaches: the genetic edition through CRISPR/Cas9 technology of genes encoding STING or cGAS in NIH/3T3 murine fibroblasts and the infection of HEK293 and HEK293 T human epithelial cells, deficient in cGAS and in cGAS and STING expression, respectively. Overall, our results suggest the existence of two different pathways involved in the establishment of the antiviral response, both dependent on STING expression. Particularly, the cGAS-STING pathway resulted in the more relevant production of beta interferon (IFN-β) and IFN-λ1 in response to baculovirus infection. In human epithelial cells, IFN-λ1 production was also induced in a cGAS-independent and DNA-protein kinase (DNA-PK)-dependent manner. Finally, we demonstrated that these cellular responses toward baculovirus infection affect the efficiency of transduction of baculovirus vectors.
IMPORTANCE Baculoviruses are nonpathogenic viruses that infect mammals, which, among other applications, are used as vehicles for gene delivery. Here, we demonstrated that the cytosolic DNA sensor cGAS recognizes baculoviral DNA and that the cGAS-STING axis is primarily responsible for the attenuation of transduction in human and mouse cell lines through type I and type III IFNs. Furthermore, we identified DNA-dependent protein kinase (DNA-PK) as a cGAS-independent and alternative DNA cytosolic sensor that contributes less to the antiviral state in baculovirus infection in human epithelial cells than cGAS. Knowledge of the pathways involved in the response of mammalian cells to baculovirus infection will improve the use of this vector as a tool for gene therapy. |
| format | Artículo |
| id | INTA10117 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | American Society for Microbiology |
| publisherStr | American Society for Microbiology |
| record_format | dspace |
| spelling | INTA101172021-08-25T18:13:33Z Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway Amalfi, Sabrina Molina, Guido Nicolas Bevacqua, Romina Jimena Lopez, Maria Gabriela Taboga, Oscar Alberto Alfonso, Victoria Baculovirus Interferonas Autographa californica Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas Edición de Genes Interferons CRISPR Gene Editing The baculovirus Autographa californica multiple nucleopolyhedrovirus is an insect virus with a circular double-stranded DNA genome, which, among other multiple biotechnological applications, is used as an expression vector for gene delivery in mammalian cells. Nevertheless, the nonspecific immune response triggered by viral vectors often suppresses transgene expression. To understand the mechanisms involved in that response, in the present study, we studied the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway by using two approaches: the genetic edition through CRISPR/Cas9 technology of genes encoding STING or cGAS in NIH/3T3 murine fibroblasts and the infection of HEK293 and HEK293 T human epithelial cells, deficient in cGAS and in cGAS and STING expression, respectively. Overall, our results suggest the existence of two different pathways involved in the establishment of the antiviral response, both dependent on STING expression. Particularly, the cGAS-STING pathway resulted in the more relevant production of beta interferon (IFN-β) and IFN-λ1 in response to baculovirus infection. In human epithelial cells, IFN-λ1 production was also induced in a cGAS-independent and DNA-protein kinase (DNA-PK)-dependent manner. Finally, we demonstrated that these cellular responses toward baculovirus infection affect the efficiency of transduction of baculovirus vectors. IMPORTANCE Baculoviruses are nonpathogenic viruses that infect mammals, which, among other applications, are used as vehicles for gene delivery. Here, we demonstrated that the cytosolic DNA sensor cGAS recognizes baculoviral DNA and that the cGAS-STING axis is primarily responsible for the attenuation of transduction in human and mouse cell lines through type I and type III IFNs. Furthermore, we identified DNA-dependent protein kinase (DNA-PK) as a cGAS-independent and alternative DNA cytosolic sensor that contributes less to the antiviral state in baculovirus infection in human epithelial cells than cGAS. Knowledge of the pathways involved in the response of mammalian cells to baculovirus infection will improve the use of this vector as a tool for gene therapy. Instituto de Biotecnología Fil: Amalfi, Sabrina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Amalfi, Sabrina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Molina, Guido Nicolas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Molina, Guido Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Bevacqua, Romina Jimena. Universidad de Buenos Aires. Facultad de Agronomía. Laboratorio de Biotecnología Animal; Argentina Fil: Bevacqua, Romina Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Bevacqua, Romina Jimena. Stanford University School of Medicine. Department of Developmental Biology. Seung Kim Lab; Estados Unidos Fil: Lopez, Maria Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Lopez, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Taboga, Oscar Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Alfonso, Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Alfonso, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2021-08-25T17:58:49Z 2021-08-25T17:58:49Z 2020-11 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/10117 https://journals.asm.org/doi/10.1128/JVI.01555-20 0022-538X https://doi.org/10.1128/JVI.01555-20 eng info:eu-repograntAgreement/INTA/PNBIO-1131034/AR./Inmunología molecular y genómica funcional aplicadas a interacciones patógeno hospedador de interés pecuario. info:eu-repo/semantics/restrictedAccess application/pdf American Society for Microbiology Journal of Virology 94 (21) : e01555-20 (Noviembre 2020) |
| spellingShingle | Baculovirus Interferonas Autographa californica Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas Edición de Genes Interferons CRISPR Gene Editing Amalfi, Sabrina Molina, Guido Nicolas Bevacqua, Romina Jimena Lopez, Maria Gabriela Taboga, Oscar Alberto Alfonso, Victoria Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway |
| title | Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway |
| title_full | Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway |
| title_fullStr | Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway |
| title_full_unstemmed | Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway |
| title_short | Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway |
| title_sort | baculovirus transduction in mammalian cells is affected by the production of type i and iii interferons which is mediated mainly by the cgas sting pathway |
| topic | Baculovirus Interferonas Autographa californica Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas Edición de Genes Interferons CRISPR Gene Editing |
| url | http://hdl.handle.net/20.500.12123/10117 https://journals.asm.org/doi/10.1128/JVI.01555-20 https://doi.org/10.1128/JVI.01555-20 |
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