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Unité de Biologie Moléculaire du Gène, Institut National de la Santé et de la Recherche Médicale Unité 277, Institut Pasteur, Paris, France;
Unité d’Immunologie et d’Immunothérapie, Institut Gustave Roussy, Villejuif, France; and
Virogenetics, Troy, NY 12180
We have investigated the possible usefulness of recombinant
canarypox virus (ALVAC) encoding the melanoma-associated Ag,
Melan-A/MART-1 (MART-1), in cancer immunotherapy, using a dendritic
cell (DC)-based approach. ALVAC MART-1-infected DC express, and are
able to process and present, the Ag coded by the viral vector. One
consistent feature of infection by ALVAC is that these viruses induce
apoptosis, and we show cross-presentation of Ag when uninfected DC are
cocultured with ALVAC MART-1-infected DC. Uptake of apoptotic virally
infected DC by uninfected DC and subsequent expression of tumor Ag in
the latter were verified by flow cytometry analysis, image cytometry,
and confocal microscopy. Functional activity was monitored in vitro by
the stimulation of a MART-1-specific cytotoxic T cell clone. Heightened
efficiency in Ag presentation is evidenced in the 2- to 3-fold increase
in IFN-
production by the T cell clone, as compared with the
ALVAC-infected DC alone. Cocultures of ALVAC MART-1-infected and
uninfected DC are able to induce MART-1-specific T cell immune
responses, as assessed by HLA class I/peptide tetramer binding, IFN-
ELISPOT assays, and cytotoxicity tests. Overall, our data indicate that
DC infected with recombinant canarypox viruses may represent an
efficient presentation platform for tumor Ags, which can be exploited
in clinical studies.
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