title: Improved innate and adaptive immunostimulation by genetically modified HIV-1 protein expressing NYVAC vectors.
abstract: Attenuated poxviruses are safe and capable of expressing foreign antigens. Poxviruses are applied in veterinary vaccination and explored as candidate vaccines for humans. However, poxviruses express multiple genes encoding proteins that interfere with components of the innate and adaptive immune response. This manuscript describes two strategies aimed to improve the immunogenicity of the highly attenuated, host-range restricted poxvirus NYVAC: deletion of the viral gene encoding type-I interferon-binding protein and development of attenuated replication-competent NYVAC. We evaluated these newly generated NYVAC mutants, encoding HIV-1 env, gag, pol and nef, for their ability to stimulate HIV-specific CD8 T-cell responses in vitro from blood mononuclear cells of HIV-infected subjects. The new vectors were evaluated and compared to the parental NYVAC vector in dendritic cells (DCs), RNA expression arrays, HIV gag expression and cross-presentation assays in vitro. Deletion of type-I interferon-binding protein enhanced expression of interferon and interferon-induced genes in DCs, and increased maturation of infected DCs. Restoration of replication competence induced activation of pathways involving antigen processing and presentation. Also, replication-competent NYVAC showed increased Gag expression in infected cells, permitting enhanced cross-presentation to HIV-specific CD8 T cells and proliferation of HIV-specific memory CD8 T-cells in vitro. The recombinant NYVAC combining both modifications induced interferon-induced genes and genes involved in antigen processing and presentation, as well as increased Gag expression. This combined replication-competent NYVAC is a promising candidate for the next generation of HIV vaccines.
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Esther D
authors.initials: ED
authors.lastname: Quakkelaar
authors.forename: Anke
authors.initials: A
authors.lastname: Redeker
authors.forename: Elias K
authors.initials: EK
authors.lastname: Haddad
authors.forename: Alexandre
authors.initials: A
authors.lastname: Harari
authors.forename: Stella Mayo
authors.initials: SM
authors.lastname: McCaughey
authors.forename: Thomas
authors.initials: T
authors.lastname: Duhen
authors.forename: Abdelali
authors.initials: A
authors.lastname: Filali-Mouhim
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Goulet
authors.forename: Nikki M
authors.initials: NM
authors.lastname: Loof
authors.forename: Ferry
authors.initials: F
authors.lastname: Ossendorp
authors.forename: Beatriz
authors.initials: B
authors.lastname: Perdiguero
authors.forename: Paul
authors.initials: P
authors.lastname: Heinen
authors.forename: Carmen E
authors.initials: CE
authors.lastname: Gomez
authors.forename: Karen V
authors.initials: KV
authors.lastname: Kibler
authors.forename: David M
authors.initials: DM
authors.lastname: Koelle
authors.forename: Rafick P
authors.initials: RP
authors.lastname: Sékaly
authors.forename: Federica
authors.initials: F
authors.lastname: Sallusto
authors.forename: Antonio
authors.initials: A
authors.lastname: Lanzavecchia
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Pantaleo
authors.forename: Mariano
authors.initials: M
authors.lastname: Esteban
authors.forename: Jim
authors.initials: J
authors.lastname: Tartaglia
authors.forename: Bertram L
authors.initials: BL
authors.lastname: Jacobs
authors.forename: Cornelis J M
authors.initials: CJ
authors.lastname: Melief
mesh_terms: D056704:Adaptive Immunity / Q000235:genetics*; D000818:Animals; D017951:Antigen Presentation / Q000235:genetics; D051940:B7-2 Antigen / Q000378:metabolism; D049109:Cell Proliferation; D006224:Cricetinae; D003429:Cross Reactions / Q000235:genetics; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism / Q000821:virology; D017353:Gene Deletion; D015870:Gene Expression; D005786:Gene Expression Regulation / Q000235:genetics / Q000276:immunology; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D007113:Immunity, Innate / Q000235:genetics*; D007114:Immunization / Q000379:methods*; D016898:Interferon-alpha / Q000096:biosynthesis; D011212:Poxviridae / Q000235:genetics* / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000821:virology; D014764:Viral Proteins / Q000235:genetics*; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000235:genetics
chemical_list: D051940:B7-2 Antigen; D016898:Interferon-alpha; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus