dceab64c583c5bc74c6fa651b4d90b72 pmc_10934719.pdf 17ed167c45c6e3a5caf1daa9ac385c1d8053975f pmc_10934719.pdf 794d709a20fd7139883d26a2d988cd25b4d74abb3c43b7c6d5330be145f1e91b pmc_10934719.pdf Title: Nanoparticle-Based Secretory Granules Induce a Specific and Long-Lasting Immune Response through Prolonged Antigen Release Subject: Developing prolonged antigen delivery systems that mimic long-term exposure to pathogens appears as a promising but still poorly explored approach to reach durable immunities. In this study, we have used a simple technology by which His-tagged proteins can be assembled, assisted by divalent cations, as supramolecular complexes with progressive complexity, namely protein-only nanoparticles and microparticles. Microparticles produced out of nanoparticles are biomimetics of secretory granules from the mammalian hormonal system. Upon subcutaneous administration, they slowly disintegrate, acting as an endocrine-like secretory system and rendering the building block nanoparticles progressively bioavailable. The performance of such materials, previously validated for drug delivery in oncology, has been tested here regarding the potential for time-prolonged antigen release. This has been completed by taking, as a building block, a nanostructured version of p30, a main structural immunogen from the African swine fever virus (ASFV). By challenging the system in both mice and pigs, we have observed unusually potent pro-inflammatory activity in porcine macrophages, and long-lasting humoral and cellular responses in vivo, which might overcome the need for an adjuvant. The robustness of both innate and adaptive responses tag, for the first time, these dynamic depot materials as a novel and valuable instrument with transversal applicability in immune stimulation and vaccinology. Keywords: recombinant proteins; biomaterials; protein nanoparticles; secretory granules; immunization Author: Laia Bosch-Camós, Carlos Martínez-Torró, Hèctor López-Laguna, Jara Lascorz, Jordi Argilaguet, Antonio Villaverde, Fernando Rodríguez and Esther Vázquez Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Thu Feb 29 02:34:34 2024 CET ModDate: Thu Feb 29 03:56:20 2024 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 16 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4387151 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- HFBUVF+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 MDSFBA+URWPalladioL-Bold Type 1 Custom yes yes yes 16 0 QKYOXM+URWPalladioL-Roma Type 1 Custom yes yes yes 22 0 XNEXOK+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 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pmc_10934719.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-03-20 02:30:34 CET Size: 4387151 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 1092 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Ethics Statement Destination: subsection.2.1 Item: Title: Protein Design, Production and Purification Destination: subsection.2.2 Item: Title: Dynamic Light Scattering Destination: subsection.2.3 Item: Title: Microparticle Formation and Protein Release Destination: subsection.2.4 Item: Title: Scanning Electron Microscopy Destination: subsection.2.5 Item: Title: Study Design of the Pig Experiments Destination: subsection.2.6 Item: Title: Study Design of the Mice Experiment Destination: subsection.2.7 Item: Title: ELISA Destination: subsection.2.8 Item: Title: Mouse Splenocytes Collection and Flow Cytometry Destination: subsection.2.9 Item: Title: ELISpot Assay with Porcine Peripheral Blood Monocyte Cells (PBMCs) Destination: subsection.2.10 Item: Title: Isolation of Porcine Alveolar Macrophages (PAMs) Destination: subsection.2.11 Item: Title: Multiplex Luminex Assay Destination: subsection.2.12 Item: Title: Statistical Analyses Destination: subsection.2.13 Item: Title: Results Destination: section.3 Children: Item: Title: Design and Construction of an ASFV-Antigen Nanoparticle Destination: subsection.3.1 Item: Title: RK4-p30-H6 Microparticles Promote Consistent and Prolonged Antigen Release Destination: subsection.3.2 Item: Title: Subcutaneous Administration of p30 POMs Are Safe and Immunogenic in Pigs Destination: subsection.3.3 Item: Title: POM Proteins Stimulate the Immune System in a Non-Specific Manner Destination: subsection.3.4 Item: Title: Subcutaneous Administration of GFP POMs Are Safe and Immunogenic in Mice Destination: subsection.3.5 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.6 Info: Title: Nanoparticle-Based Secretory Granules Induce a Specific and Long-Lasting Immune Response through Prolonged Antigen Release Author: Laia Bosch-Camós, Carlos Martínez-Torró, Hèctor López-Laguna, Jara Lascorz, Jordi Argilaguet, Antonio Villaverde, Fernando Rodríguez and Esther Vázquez Subject: Developing prolonged antigen delivery systems that mimic long-term exposure to pathogens appears as a promising but still poorly explored approach to reach durable immunities. In this study, we have used a simple technology by which His-tagged proteins can be assembled, assisted by divalent cations, as supramolecular complexes with progressive complexity, namely protein-only nanoparticles and microparticles. Microparticles produced out of nanoparticles are biomimetics of secretory granules from the mammalian hormonal system. Upon subcutaneous administration, they slowly disintegrate, acting as an endocrine-like secretory system and rendering the building block nanoparticles progressively bioavailable. The performance of such materials, previously validated for drug delivery in oncology, has been tested here regarding the potential for time-prolonged antigen release. This has been completed by taking, as a building block, a nanostructured version of p30, a main structural immunogen from the African swine fever virus (ASFV). By challenging the system in both mice and pigs, we have observed unusually potent pro-inflammatory activity in porcine macrophages, and long-lasting humoral and cellular responses in vivo, which might overcome the need for an adjuvant. The robustness of both innate and adaptive responses tag, for the first time, these dynamic depot materials as a novel and valuable instrument with transversal applicability in immune stimulation and vaccinology. 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