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Abstract
Background
In vivo electroporation (EP) has proven to significantly increase transfection efficiency
and augment immune responses to plasmid DNA encoded vaccine antigens. In mice, we
have attempted to establish the optimal EP immunization protocol for delivering HIV-1
antigens.
Methods
Mice were immunized with a plasmid encoding HIV-1 gag p37, either intradermally (id)
with the DermaVax EP device, or intramuscularly (im) without EP. The different parameters
explored were; the number of immunizations, the interval between immunizations, and
the choice of priming (im or id). A novel Fluorospot assay was used to evaluate the
vaccine specific cellular immune responses. This assay allows for detection of vaccine-specific
cells that secrete IFN-γ, IL-2 and cells that simultaneously secrete both cytokines.
In order to verify the Fluorospot results, conventional ELISpot assay was used. The
humoral vaccine-specific response was evaluated by ELISA.
Results
The main findings were: 1) two and three id+EP immunizations induced similar and high
cellular (up to 8000 IFN-γ SFC/million splenocytes) responses while antibody responses
were increased after three as compared to two immunizations, 2) one month interval
between immunizations was superior to two months in terms of cellular responses, and
3) repeated id+EP immunizations induced higher immune responses than im priming followed
by id+EP boost.
Conclusion
The Fluorospot and the ELIspot assays gave similar results, confirming the sensitivity
of the Fluorospot assay. Moreover, the Fluorospot assay enabled identification of
high quality vaccine-specific cells simultaneously secreting both IFN-γ and IL-2.
Three id+EP immunizations induced the highest levels of antibodies, while two immunizations
were sufficient to induce a strong cellular response. In an upcoming longitudinal
study we will further investigate the capacity of id EP to induce long lasting immune
responses and also investigate the qualitative differences in responses induced by
single vs. multiple immunizations.