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* Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164;
Department of Animal Sciences, University of Illinois, Urbana-Champaign, IL 61801; and
Compton Laboratory, Institute of Animal Health, Compton, Newbury, United Kingdom
DNA-based immunization is a contemporary strategy for developing
vaccines to prevent infectious diseases in animals and humans.
Translating the efficacy of DNA immunization demonstrated in murine
models to the animal species that represent the actual populations to
be protected remains a significant challenge. We tested two hypotheses
directed at enhancing DNA vaccine efficacy in outbred animals. The
first hypothesis, that DNA-encoding fetal liver tyrosine kinase 3
ligand (Flt3L) and GM-CSF increases dendritic cell (DC) recruitment to
the immunization site, was tested by intradermal inoculation of calves
with plasmid DNA encoding Flt3L and GM-CSF followed by quantitation of
CD1+ DC. Peak DC recruitment was detected at 10–15 days
postinoculation and was significantly greater (p <
0.05) in calves in the treatment group as compared with control calves
inoculated identically, but without Flt3L and GM-CSF. The second
hypothesis, that DNA encoding Flt3L and GM-CSF enhances immunity to a
DNA vector-expressed Ag, was tested by analyzing the CD4+ T
lymphocyte response to Anaplasma marginale major surface
protein 1a (MSP1a). Calves immunized with DNA-expressing MSP1a
developed strong CD4+ T cell responses against A.
marginale, MSP1a, and specific MHC class II DR-restricted MSP1a
epitopes. Administration of DNA-encoding Flt3L and GM-CSF before MSP1a
DNA vaccination significantly increased the population of Ag-specific
effector/memory cells in PBMC and significantly enhanced MSP1a-specific
CD4+ T cell proliferation and IFN-
secretion as compared
with MHC class II DR-matched calves vaccinated
identically but without Flt3L and GM-CSF. These results support use of
these growth factors in DNA vaccination and specifically indicate their
applicability for vaccine testing in outbred
animals.
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