| Summary: | 碩士 === 國立中興大學 === 獸醫學系暨研究所 === 98 === Melanoma cells selectively express a number of differentiation antigens including melan-A, gp100, and tyrosinase that can serve as targets for an immunotherapy strategy. The aim of our study was to construct a melanoma specific antigen expression plasmid as an immunotherapy for canine melanoma. A multiepitope sequence, which modified from a previous study, was generated by annealing of a set of 8 primers by PCR reaction. A plasmid in which expression of the multiepitope sequence of melanoma antigens or fused with the GFP (green fluorescence protein) was constructed. The transient expression of multiepitope-GFP in eukaryotic cells (HEK-293T) was confirmed by fluorescent microscopy and western blot analysis. Evaluation of safety and acute toxicity was performed, and adverse effect was not significant in C57BL6/J mice intramuscularly injected with multiepitope plasmid at three different doses (5, 50, 250 μg) after 7 days of observation. Furthermore, a melanoma model was successfully established in C57BL/6J mice and the therapeutic effect of the plasmid was evaluated on these tumor bearing mice. The growth of tumor volume in the therapeutic group was significant inhibited than the mock group (＊p<0.05) and control group (＊p<0.05) on day 24. The multiepitope plasmid might be a potential therapy to regress the melanoma or improve survival time of patients and further investigation of multiepitope plasmid on canine melanoma is necessary.
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