title: Preparation of clinical-grade recombinant canarypox-human immunodeficiency virus vaccine-loaded human dendritic cells.
abstract: Preclinical data are reported that support a human immunodeficiency virus (HIV) vaccine strategy using recombinant canarypox-HIV vectors (ALVAC-HIV) to load human dendritic cells (DCs) with HIV antigens. Clinical-grade DCs were infected with good manufacturing practice-grade ALVAC-HIV vaccine constructs. ALVAC infection, HIV gene expression, and DC viability and function were monitored by use of immunohistochemistry, flow cytometry, blastogenesis assays, antigen-specific interferon (IFN)-gamma enzyme-linked immunospot assay, and enzyme-linked immunosorbent assay protein detection. The vaccines infected both immature and mature DCs, and intracellular HIV-1 Gag protein was detected within hours. ALVAC-HIV induced DC maturation that was mediated by tumor necrosis factor-alpha and induced DC apoptosis that was directly related to the length of vaccine exposure. Of importance, the infected DCs remained functional in T cell stimulation assays and induced HIV antigen-specific CD8(+) T cell production of IFN-gamma from cells of HIV-1-infected individuals. These data support an ongoing HIV vaccine trial comparing conventional vaccine delivery routes with ex vivo vaccine-loaded autologous DCs for immunogenicity in HIV-1-uninfected volunteers.
authors.affiliation: Division of Retrovirology, US Military HIV Research Program, Rockville, Maryland 20850, USA. mmarovich@hivresearch.org
authors.forename: Mary A
authors.initials: MA
authors.lastname: Marovich
authors.forename: John R
authors.initials: JR
authors.lastname: Mascola
authors.forename: Michael A
authors.initials: MA
authors.lastname: Eller
authors.forename: Mark K
authors.initials: MK
authors.lastname: Louder
authors.forename: Pierre A
authors.initials: PA
authors.lastname: Caudrelier
authors.forename: Raphaelle
authors.initials: R
authors.lastname: El-Habib
authors.forename: Silvia
authors.initials: S
authors.lastname: Ratto-Kim
authors.forename: Josephine H
authors.initials: JH
authors.lastname: Cox
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Currier
authors.forename: Bruce L
authors.initials: BL
authors.lastname: Levine
authors.forename: Carl H
authors.initials: CH
authors.lastname: June
authors.forename: Wendy B
authors.initials: WB
authors.lastname: Bernstein
authors.forename: Merlin L
authors.initials: ML
authors.lastname: Robb
authors.forename: Beatrice
authors.initials: B
authors.lastname: Schuler-Thurner
authors.forename: Ralph M
authors.initials: RM
authors.lastname: Steinman
authors.forename: Deborah L
authors.initials: DL
authors.lastname: Birx
authors.forename: Sarah
authors.initials: S
authors.lastname: Schlesinger-Frankel
mesh_terms: D016915:AIDS Vaccines / Q000276:immunology*; D015703:Antigens, CD / Q000032:analysis; D028121:Canarypox virus / Q000276:immunology*; D002469:Cell Separation / Q000379:methods; D003713:Dendritic Cells / Q000276:immunology*; D015683:Gene Products, gag / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D008213:Lymphocyte Activation / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000276:immunology*; D014765:Viral Vaccines / Q000138:chemical synthesis / Q000276:immunology
chemical_list: D016915:AIDS Vaccines; C078788:ALVAC vaccine; D015703:Antigens, CD; D015683:Gene Products, gag; D014614:Vaccines, Synthetic; D014765:Viral Vaccines