f33bc853a6539e85f8c3eb89e6506339 pmc_10219511.pdf 8ac315b06923cbbf5450f3649f32c8c39360939b pmc_10219511.pdf c469bea40ab71d2a14c5f869e93d6a371a1bb5b160b6b20a72f1947484fa01b5 pmc_10219511.pdf Title: Exploring FeLV-Gag-Based VLPs as a New Vaccine Platform—Analysis of Production and Immunogenicity Subject: Feline leukemia virus (FeLV) is one of the most prevalent infectious diseases in domestic cats. Although different commercial vaccines are available, none of them provides full protection. Thus, efforts to design a more efficient vaccine are needed. Our group has successfully engineered HIV-1 Gag-based VLPs that induce a potent and functional immune response against the HIV-1 transmembrane protein gp41. Here, we propose to use this concept to generate FeLV-Gag-based VLPs as a novel vaccine strategy against this retrovirus. By analogy to our HIV-1 platform, a fragment of the FeLV transmembrane p15E protein was exposed on FeLV-Gag-based VLPs. After optimization of Gag sequences, the immunogenicity of the selected candidates was evaluated in C57BL/6 and BALB/c mice, showing strong cellular and humoral responses to Gag but failing to generate anti-p15E antibodies. Altogether, this study not only tests the versatility of the enveloped VLP-based vaccine platform but also sheds light on FeLV vaccine research. Keywords: FeLV vaccination; virus-like particle; humoral immunity; glycogag; veterinary science Author: Raquel Ortiz, Ana Barajas, Anna Pons-Grífols, Benjamin Trinité, Ferran Tarrés-Freixas, Carla Rovirosa, Victor Urrea, Antonio Barreiro, Anna Gonzalez-Tendero, Maria Cardona, Laura Ferrer, Bonaventura Clotet, Jorge Carrillo, Carmen Aguilar-Gurrieri and Julià Blanco Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon May 22 05:58:50 2023 CEST ModDate: Mon May 22 06:10:22 2023 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 22 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3842551 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- XXEPLQ+VnURWPalladioL Type 1 Custom yes yes no 10 0 EJEPSV+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 FUVCGH+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 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WinAnsi yes yes no 168 0 FHDBPP+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 171 0 FHDELP+PalatinoLinotype TrueType MacRoman yes yes no 174 0 FHDBOL+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 202 0 FHDBON+PalatinoLinotype TrueType WinAnsi yes yes no 205 0 FHDBPP+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 208 0 FHDELP+PalatinoLinotype TrueType MacRoman yes yes no 211 0 KSJIQB+TeX-tipa10 Type 1 Builtin yes yes no 221 0 OQIVQT+EURM10 Type 1 Builtin yes yes no 225 0 JOXUTR+TeX-tipa9 Type 1 Builtin yes yes no 232 0 FHDBOL+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 243 0 FHDBOM+PalatinoLinotype CID TrueType Identity-H yes yes yes 246 0 FHDBON+PalatinoLinotype TrueType WinAnsi yes yes no 252 0 FHDBPP+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 255 0 FHEFIJ+TimesNewRoman CID TrueType Identity-H yes yes yes 258 0 IXLVLQ+CMSY10 Type 1 Builtin yes yes no 268 0 FHDBOL+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 279 0 FHDBOM+PalatinoLinotype CID TrueType Identity-H yes yes yes 282 0 FHDBON+PalatinoLinotype TrueType WinAnsi yes yes no 288 0 FHDBPP+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 291 0 FHDELP+PalatinoLinotype TrueType MacRoman yes yes no 294 0 FHEILH+MSGothic CID TrueType Identity-H yes yes yes 297 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-09-02 02:27:57 CEST RepresentationInformation: pmc_10219511.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-07-27 02:24:16 CEST Size: 3842551 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 626 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: Generation of HIV and FeLV-Based Virus-like Particles Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Sequential Deletion of Gag to Reduce Proteolytic Cleavage Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Improving FeLV-Based VLP Production Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Immunogenicity of TM-FeLV VLPs in C57BL/6 Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Immunogenicity of TM-FeLV VLPs in BALB/c Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Children: Item: Title: Plasmids Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Cell Line, Culture Conditions, and Transfection Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Cell Disruption, VLP Extraction, and Purification Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Analysis of VLP and Protein Production Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Children: Item: Title: Western Blotting Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Flow Cytometry Analysis of VLP Expression Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Enzyme-Linked Immunosorbent Assay (ELISA) for VLP Quantification Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Transmission (TEM) and Cryo-Transmission Electron Microscopy (Cryo-EM) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Nanoparticle Tracking Analysis (NTA) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Mice Immunization and Immunogenicity Analyses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Evaluation of Humoral Response by ELISA Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Evaluation of Cellular Immune Responses by ELISpot Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Evaluation of Anti-Expi293 Humoral Responses by Flow Cytometry Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Info: Title: Exploring FeLV-Gag-Based VLPs as a New Vaccine Platform Analysis of Production and Immunogenicity Author: Raquel Ortiz, Ana Barajas, Anna Pons-Grífols, Benjamin Trinité, Ferran Tarrés-Freixas, Carla Rovirosa, Victor Urrea, Antonio Barreiro, Anna Gonzalez-Tendero, Maria Cardona, Laura Ferrer, Bonaventura Clotet, Jorge Carrillo, Carmen Aguilar-Gurrieri and Julià Blanco Subject: Feline leukemia virus (FeLV) is one of the most prevalent infectious diseases in domestic cats. Although different commercial vaccines are available, none of them provides full protection. Thus, efforts to design a more efficient vaccine are needed. Our group has successfully engineered HIV-1 Gag-based VLPs that induce a potent and functional immune response against the HIV-1 transmembrane protein gp41. Here, we propose to use this concept to generate FeLV-Gag-based VLPs as a novel vaccine strategy against this retrovirus. By analogy to our HIV-1 platform, a fragment of the FeLV transmembrane p15E protein was exposed on FeLV-Gag-based VLPs. After optimization of Gag sequences, the immunogenicity of the selected candidates was evaluated in C57BL/6 and BALB/c mice, showing strong cellular and humoral responses to Gag but failing to generate anti-p15E antibodies. Altogether, this study not only tests the versatility of the enveloped VLP-based vaccine platform but also sheds light on FeLV vaccine research. 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