Design and Functional Characterization of HIV-1 Envelope Protein-Coupled T Helper Liposomes
Functionalization of experimental HIV-1 virus-like particle vaccines with heterologous T helper epitopes (T helper VLPs) can modulate the humoral immune response via intrastructural help (ISH). Current advances in the conjugation of native-like HIV-1 envelope trimers (Env) onto liposomes and encapsu...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-06-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/14/7/1385 |
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| author | Dominik Damm Ehsan Suleiman Hannah Theobald Jannik T. Wagner Mirjam Batzoni Bianca Ahlfeld (née Kohlhauser) Bernd Walkenfort Jens-Christian Albrecht Jidnyasa Ingale Lifei Yang Mike Hasenberg Richard T. Wyatt Karola Vorauer-Uhl Klaus Überla Vladimir Temchura |
| author_facet | Dominik Damm Ehsan Suleiman Hannah Theobald Jannik T. Wagner Mirjam Batzoni Bianca Ahlfeld (née Kohlhauser) Bernd Walkenfort Jens-Christian Albrecht Jidnyasa Ingale Lifei Yang Mike Hasenberg Richard T. Wyatt Karola Vorauer-Uhl Klaus Überla Vladimir Temchura |
| author_sort | Dominik Damm |
| collection | DOAJ |
| description | Functionalization of experimental HIV-1 virus-like particle vaccines with heterologous T helper epitopes (T helper VLPs) can modulate the humoral immune response via intrastructural help (ISH). Current advances in the conjugation of native-like HIV-1 envelope trimers (Env) onto liposomes and encapsulation of peptide epitopes into these nanoparticles renders this GMP-scalable liposomal platform a feasible alternative to VLP-based vaccines. In this study, we designed and analyzed customizable Env-conjugated T helper liposomes. First, we passively encapsulated T helper peptides into a well-characterized liposome formulation displaying a dense array of Env trimers on the surface. We confirmed the closed pre-fusion state of the coupled Env trimers by immunogold staining with conformation-specific antibodies. These peptide-loaded Env-liposome conjugates efficiently activated Env-specific B cells, which further induced proliferation of CD4+ T cells by presentation of liposome-derived peptides on MHC-II molecules. The peptide encapsulation process was then quantitatively improved by an electrostatically driven approach using an overall anionic lipid formulation. We demonstrated that peptides delivered by liposomes were presented by DCs in secondary lymphoid organs after intramuscular immunization of mice. UFO (uncleaved prefusion optimized) Env trimers were covalently coupled to peptide-loaded anionic liposomes by His-tag/NTA(Ni) interactions and EDC/Sulfo-NHS crosslinking. EM imaging revealed a moderately dense array of well-folded Env trimers on the liposomal surface. The conformation was verified by liposomal surface FACS. Furthermore, anionic Env-coupled T helper liposomes effectively induced Env-specific B cell activation and proliferation in a comparable range to T helper VLPs. Taken together, we demonstrated that T helper VLPs can be substituted with customizable and GMP-scalable liposomal nanoparticles as a perspective for future preclinical and clinical HIV vaccine applications. The functional nanoparticle characterization assays shown in this study can be applied to other systems of synthetic nanoparticles delivering antigens derived from various pathogens. |
| first_indexed | 2024-03-09T06:04:48Z |
| format | Article |
| id | doaj.art-d6bcf5265dc047d3a53880797af39283 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-09T06:04:48Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-d6bcf5265dc047d3a53880797af392832023-12-03T12:06:09ZengMDPI AGPharmaceutics1999-49232022-06-01147138510.3390/pharmaceutics14071385Design and Functional Characterization of HIV-1 Envelope Protein-Coupled T Helper LiposomesDominik Damm0Ehsan Suleiman1Hannah Theobald2Jannik T. Wagner3Mirjam Batzoni4Bianca Ahlfeld (née Kohlhauser)5Bernd Walkenfort6Jens-Christian Albrecht7Jidnyasa Ingale8Lifei Yang9Mike Hasenberg10Richard T. Wyatt11Karola Vorauer-Uhl12Klaus Überla13Vladimir Temchura14Institute of Clinical and Molecular Virology, Friedrich-Alexander University Erlangen-Nürnberg, 91054 Erlangen, GermanyDepartment of Biotechnology, University of Natural Resources and Life Sciences, 1190 Vienna, AustriaInstitute of Clinical and Molecular Virology, Friedrich-Alexander University Erlangen-Nürnberg, 91054 Erlangen, GermanyInstitute of Clinical and Molecular Virology, Friedrich-Alexander University Erlangen-Nürnberg, 91054 Erlangen, GermanyDepartment of Biotechnology, University of Natural Resources and Life Sciences, 1190 Vienna, AustriaDepartment of Biotechnology, University of Natural Resources and Life Sciences, 1190 Vienna, AustriaElectron Microscopy Unit (EMU), Imaging Center Essen (IMCES), Faculty of Medicine, University of Duisburg-Essen, 45147 Essen, GermanyInstitute of Clinical and Molecular Virology, Friedrich-Alexander University Erlangen-Nürnberg, 91054 Erlangen, GermanyVaccine Business Unit, Takeda Pharmaceuticals, Cambridge, MA 02139, USADepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA 92037, USAElectron Microscopy Unit (EMU), Imaging Center Essen (IMCES), Faculty of Medicine, University of Duisburg-Essen, 45147 Essen, GermanyDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA 92037, USADepartment of Biotechnology, University of Natural Resources and Life Sciences, 1190 Vienna, AustriaInstitute of Clinical and Molecular Virology, Friedrich-Alexander University Erlangen-Nürnberg, 91054 Erlangen, GermanyInstitute of Clinical and Molecular Virology, Friedrich-Alexander University Erlangen-Nürnberg, 91054 Erlangen, GermanyFunctionalization of experimental HIV-1 virus-like particle vaccines with heterologous T helper epitopes (T helper VLPs) can modulate the humoral immune response via intrastructural help (ISH). Current advances in the conjugation of native-like HIV-1 envelope trimers (Env) onto liposomes and encapsulation of peptide epitopes into these nanoparticles renders this GMP-scalable liposomal platform a feasible alternative to VLP-based vaccines. In this study, we designed and analyzed customizable Env-conjugated T helper liposomes. First, we passively encapsulated T helper peptides into a well-characterized liposome formulation displaying a dense array of Env trimers on the surface. We confirmed the closed pre-fusion state of the coupled Env trimers by immunogold staining with conformation-specific antibodies. These peptide-loaded Env-liposome conjugates efficiently activated Env-specific B cells, which further induced proliferation of CD4+ T cells by presentation of liposome-derived peptides on MHC-II molecules. The peptide encapsulation process was then quantitatively improved by an electrostatically driven approach using an overall anionic lipid formulation. We demonstrated that peptides delivered by liposomes were presented by DCs in secondary lymphoid organs after intramuscular immunization of mice. UFO (uncleaved prefusion optimized) Env trimers were covalently coupled to peptide-loaded anionic liposomes by His-tag/NTA(Ni) interactions and EDC/Sulfo-NHS crosslinking. EM imaging revealed a moderately dense array of well-folded Env trimers on the liposomal surface. The conformation was verified by liposomal surface FACS. Furthermore, anionic Env-coupled T helper liposomes effectively induced Env-specific B cell activation and proliferation in a comparable range to T helper VLPs. Taken together, we demonstrated that T helper VLPs can be substituted with customizable and GMP-scalable liposomal nanoparticles as a perspective for future preclinical and clinical HIV vaccine applications. The functional nanoparticle characterization assays shown in this study can be applied to other systems of synthetic nanoparticles delivering antigens derived from various pathogens.https://www.mdpi.com/1999-4923/14/7/1385T helper liposomesEnv trimercoupling mechanismpeptide encapsulationintrastructural helpHIV |
| spellingShingle | Dominik Damm Ehsan Suleiman Hannah Theobald Jannik T. Wagner Mirjam Batzoni Bianca Ahlfeld (née Kohlhauser) Bernd Walkenfort Jens-Christian Albrecht Jidnyasa Ingale Lifei Yang Mike Hasenberg Richard T. Wyatt Karola Vorauer-Uhl Klaus Überla Vladimir Temchura Design and Functional Characterization of HIV-1 Envelope Protein-Coupled T Helper Liposomes Pharmaceutics T helper liposomes Env trimer coupling mechanism peptide encapsulation intrastructural help HIV |
| title | Design and Functional Characterization of HIV-1 Envelope Protein-Coupled T Helper Liposomes |
| title_full | Design and Functional Characterization of HIV-1 Envelope Protein-Coupled T Helper Liposomes |
| title_fullStr | Design and Functional Characterization of HIV-1 Envelope Protein-Coupled T Helper Liposomes |
| title_full_unstemmed | Design and Functional Characterization of HIV-1 Envelope Protein-Coupled T Helper Liposomes |
| title_short | Design and Functional Characterization of HIV-1 Envelope Protein-Coupled T Helper Liposomes |
| title_sort | design and functional characterization of hiv 1 envelope protein coupled t helper liposomes |
| topic | T helper liposomes Env trimer coupling mechanism peptide encapsulation intrastructural help HIV |
| url | https://www.mdpi.com/1999-4923/14/7/1385 |
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