Preparation of clinical-grade recombinant canarypox-human immunodeficiency virus vaccine-loaded human dendritic cells.
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.
Marovich
Mary A
MA
Division of Retrovirology, US Military HIV Research Program, Rockville, Maryland 20850, USA. mmarovich@hivresearch.org
Mascola
John R
JR
Eller
Michael A
MA
Louder
Mark K
MK
Caudrelier
Pierre A
PA
El-Habib
Raphaelle
R
Ratto-Kim
Silvia
S
Cox
Josephine H
JH
Currier
Jeffrey R
JR
Levine
Bruce L
BL
June
Carl H
CH
Bernstein
Wendy B
WB
Robb
Merlin L
ML
Schuler-Thurner
Beatrice
B
Steinman
Ralph M
RM
Birx
Deborah L
DL
Schlesinger-Frankel
Sarah
S
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
D016915:AIDS Vaccines; C078788:ALVAC vaccine; D015703:Antigens, CD; D015683:Gene Products, gag; D014614:Vaccines, Synthetic; D014765:Viral Vaccines