Establishment of an Antiplasmodial Vaccine Based on PfRH5-Encoding RNA Replicons Stabilized by Cationic Liposomes
Background: Nucleic acid-based vaccines have been studied for the past four decades, but the approval of the first messenger RNA (mRNA) vaccines during the COVID-19 pandemic opened renewed perspectives for the development of similar vaccines against different infectious diseases. Presently available...
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MDPI AG
2023-04-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/15/4/1223 |
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| author | Wesley L. Fotoran Jamile Ramos da Silva Christiane Glitz Luís Carlos de Souza Ferreira Gerhard Wunderlich |
| author_facet | Wesley L. Fotoran Jamile Ramos da Silva Christiane Glitz Luís Carlos de Souza Ferreira Gerhard Wunderlich |
| author_sort | Wesley L. Fotoran |
| collection | DOAJ |
| description | Background: Nucleic acid-based vaccines have been studied for the past four decades, but the approval of the first messenger RNA (mRNA) vaccines during the COVID-19 pandemic opened renewed perspectives for the development of similar vaccines against different infectious diseases. Presently available mRNA vaccines are based on non-replicative mRNA, which contains modified nucleosides encased in lipid vesicles, allowing for entry into the host cell cytoplasm, and reducing inflammatory reactions. An alternative immunization strategy employs self-amplifying mRNA (samRNA) derived from alphaviruses, but lacks viral structural genes. Once incorporated into ionizable lipid shells, these vaccines lead to enhanced gene expression, and lower mRNA doses are required to induce protective immune responses. In the present study, we tested a samRNA vaccine formulation based on the SP6 Venezuelan equine encephalitis (VEE) vector incorporated into cationic liposomes (dimethyldioctadecyl ammonium bromide and a cholesterol derivative). Three vaccines were generated that encoded two reporter genes (GFP and nanoLuc) and the <i>Plasmodium falciparum</i> reticulocyte binding protein homologue 5 (PfRH5). Methods: Transfection assays were performed using Vero and HEK293T cells, and the mice were immunized via the intradermal route using a tattooing device. Results: The liposome–replicon complexes showed high transfection efficiencies with in vitro cultured cells, whereas tattooing immunization with GFP-encoding replicons demonstrated gene expression in mouse skin up to 48 h after immunization. Mice immunized with liposomal PfRH5-encoding RNA replicons elicited antibodies that recognized the native protein expressed in <i>P. falciparum</i> schizont extracts, and inhibited the growth of the parasite in vitro. Conclusion: Intradermal delivery of cationic lipid-encapsulated samRNA constructs is a feasible approach for developing future malaria vaccines. |
| first_indexed | 2024-03-11T04:38:47Z |
| format | Article |
| id | doaj.art-ee619de033b04cdca6ca0fd1c88d1dff |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-11T04:38:47Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-ee619de033b04cdca6ca0fd1c88d1dff2023-11-17T20:54:27ZengMDPI AGPharmaceutics1999-49232023-04-01154122310.3390/pharmaceutics15041223Establishment of an Antiplasmodial Vaccine Based on PfRH5-Encoding RNA Replicons Stabilized by Cationic LiposomesWesley L. Fotoran0Jamile Ramos da Silva1Christiane Glitz2Luís Carlos de Souza Ferreira3Gerhard Wunderlich4Department of Parasitology, Institute for Biomedical Sciences, University of São Paulo, Avenida Professor Lineu Prestes 1374, São Paulo 05508-000, SP, BrazilDepartment of Microbiology, Institute for Biomedical Sciences, University of São Paulo, Avenida Professor Lineu Prestes 1374, São Paulo 05508-000, SP, BrazilDepartment of Molecular Physiology, Institute of Animal Physiology, Westfälische Wilhelms University of Münster, 48149 Münster, GermanyDepartment of Microbiology, Institute for Biomedical Sciences, University of São Paulo, Avenida Professor Lineu Prestes 1374, São Paulo 05508-000, SP, BrazilDepartment of Parasitology, Institute for Biomedical Sciences, University of São Paulo, Avenida Professor Lineu Prestes 1374, São Paulo 05508-000, SP, BrazilBackground: Nucleic acid-based vaccines have been studied for the past four decades, but the approval of the first messenger RNA (mRNA) vaccines during the COVID-19 pandemic opened renewed perspectives for the development of similar vaccines against different infectious diseases. Presently available mRNA vaccines are based on non-replicative mRNA, which contains modified nucleosides encased in lipid vesicles, allowing for entry into the host cell cytoplasm, and reducing inflammatory reactions. An alternative immunization strategy employs self-amplifying mRNA (samRNA) derived from alphaviruses, but lacks viral structural genes. Once incorporated into ionizable lipid shells, these vaccines lead to enhanced gene expression, and lower mRNA doses are required to induce protective immune responses. In the present study, we tested a samRNA vaccine formulation based on the SP6 Venezuelan equine encephalitis (VEE) vector incorporated into cationic liposomes (dimethyldioctadecyl ammonium bromide and a cholesterol derivative). Three vaccines were generated that encoded two reporter genes (GFP and nanoLuc) and the <i>Plasmodium falciparum</i> reticulocyte binding protein homologue 5 (PfRH5). Methods: Transfection assays were performed using Vero and HEK293T cells, and the mice were immunized via the intradermal route using a tattooing device. Results: The liposome–replicon complexes showed high transfection efficiencies with in vitro cultured cells, whereas tattooing immunization with GFP-encoding replicons demonstrated gene expression in mouse skin up to 48 h after immunization. Mice immunized with liposomal PfRH5-encoding RNA replicons elicited antibodies that recognized the native protein expressed in <i>P. falciparum</i> schizont extracts, and inhibited the growth of the parasite in vitro. Conclusion: Intradermal delivery of cationic lipid-encapsulated samRNA constructs is a feasible approach for developing future malaria vaccines.https://www.mdpi.com/1999-4923/15/4/1223nucleic acid vaccinesRNA repliconscationic liposomesmalariaPfRH5intradermal immunization |
| spellingShingle | Wesley L. Fotoran Jamile Ramos da Silva Christiane Glitz Luís Carlos de Souza Ferreira Gerhard Wunderlich Establishment of an Antiplasmodial Vaccine Based on PfRH5-Encoding RNA Replicons Stabilized by Cationic Liposomes Pharmaceutics nucleic acid vaccines RNA replicons cationic liposomes malaria PfRH5 intradermal immunization |
| title | Establishment of an Antiplasmodial Vaccine Based on PfRH5-Encoding RNA Replicons Stabilized by Cationic Liposomes |
| title_full | Establishment of an Antiplasmodial Vaccine Based on PfRH5-Encoding RNA Replicons Stabilized by Cationic Liposomes |
| title_fullStr | Establishment of an Antiplasmodial Vaccine Based on PfRH5-Encoding RNA Replicons Stabilized by Cationic Liposomes |
| title_full_unstemmed | Establishment of an Antiplasmodial Vaccine Based on PfRH5-Encoding RNA Replicons Stabilized by Cationic Liposomes |
| title_short | Establishment of an Antiplasmodial Vaccine Based on PfRH5-Encoding RNA Replicons Stabilized by Cationic Liposomes |
| title_sort | establishment of an antiplasmodial vaccine based on pfrh5 encoding rna replicons stabilized by cationic liposomes |
| topic | nucleic acid vaccines RNA replicons cationic liposomes malaria PfRH5 intradermal immunization |
| url | https://www.mdpi.com/1999-4923/15/4/1223 |
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