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Blood, Vol. 92 No. 9 (November 1), 1998:
pp. 3172-3176
Gene Transfer in Dendritic Cells, Induced by Oral DNA Vaccination
With Salmonella typhimurium, Results in Protective
Immunity Against a Murine Fibrosarcoma
Paola Paglia,
Eva Medina,
Ivano Arioli,
Carlos A. Guzman, and
Mario P. Colombo
From the Division of Experimental Oncology D, Istituto Nazionale per
Lo Studio e la Cura dei Tumori, Milano, Italy; and the Division of
Microbiology, GBF-National Research Centre for Biotechnology,
Braunschweig, Germany.
A live attenuated AroA auxotrophic mutant of
Salmonella typhimurium (SL7207) has been used as carrier for
the pCMV vector that contains the -galactosidase (-gal) gene
under the control of the immediate early promoter of
Cytomegalovirus (CMV). We tested whether orally administered
bacterial carrier could enter and deliver the transgene to
antigen-presenting cells (APCs) through the natural enteric route of
infection and whether -gal expression could generate a protective
response against an aggressive murine fibrosarcoma transduced with the
-gal gene (F1.A11) that behaves operationally as a tumor-associated
antigen. After three courses, at 15-day intervals, mice developed both
cell-mediated and systemic humoral responses to -gal. Mice
vaccinated with the Salmonella harboring pCMV, but not with
plasmid-less carrier, showed resistance to a challenge with F1.A11
cells. These experiments suggest that Salmonella-based DNA
immunization allows us to specifically target antigen expression in
vivo to APCs. To prove that the transgene is actually expressed by APCs
as a function of an eukaryotic promoter, the green fluorescent protein
(GFP) was placed under the control of either the eukariotic CMV or a
prokaryotic promoter. Using cytofluorometric analysis, GFP was detected
only in splenocytes of mice receiving a Salmonella carrier harboring
GFP under the CMV promoter. These results indicate that transgene
expression occurs because of a Salmonella-mediated gene
transfer to eukaryotic cells. Finally, approximately 19% of the
splenocytes expressed GFP. Among them, F4/80+ macrophages
and CD11cbright dendritic cells (DCs) were scored as
positive for GFP expression. Extensive work has been performed trying
to optimize the way to transfect DCs, ex vivo, with genes coding for
relevant antigens. We show here, for the first time, that DCs can be
directly and specifically transduced in vivo such to induce DNA
vaccination against tumors.
© 1998 by The American Society of Hematology.
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