HER-2/c-erbB2/neu DNA 疫苗具有抑制大量表現人類或者大鼠HER-2/neu蛋白質之老鼠腫瘤細胞生長已於動物模式被證實，然而對於HER-2/neu DNA 疫苗是否可抑制內生性表現HER-2/neu蛋白質之腫瘤細胞之生長仍不清楚。此次實驗我們首次利用HER-2/neu 之細胞外區域構築之N’-neu DNA疫苗，研究其是否能在內生性表現HER-2/neu之腫瘤動物模式中，具有抗腫瘤之能力。而從結果中發現，我們首次證實利用N’-neu DNA疫苗的確可達到有效抑制內生性表現HER-2/neu 之腫瘤生長。且此有效之作用是透過誘導HER-2/neu專一性之毒殺性CD8+ T 細胞所造成。此外此次的實驗，我們也同時利用融合之技術將N’-neu 基因融合上數種細胞激素基因，包括IL-2、IL-4、GM-CSF 評估是否能增強N’-neu DNA 疫苗誘導之抗腫瘤能力。而結果發現只有融合之N’-neu-IL-2 DNA疫苗可明顯增強N’-neu 之效果且分析其有效機制，也發現是透過增加毒殺性CD8+ T細胞之數量和活性而造成。接著，一樣的為了改善N’-neu DNA 疫苗之效力。我們此次實驗也利用合併Geldanamycin 與N’-neu 之策略，分析合併之治療效果於此次的腫瘤模式。而從結果中可發現，當利用低劑量之geldanamycin(2.5 �g)的確可提昇N’-neu 對抗腫瘤之能力，然而在高劑量時(10�g) 卻反而有抑制N’-neu 誘導之抗腫瘤免疫能力。而分析造成有效之機制結果證實, geldanamycin 是透過將HER-2/neu 蛋白質之降解而增加腫瘤的抗原性，因而導致經由N’-neu誘導之專一性毒殺CD8+ T細胞增加認識而毒殺腫瘤細胞之機會。最後，為改善N’-neu DNA疫苗之抗腫瘤能力，我們也試著利用不同方式投遞DNA之策略來評估。而從結果中我們發現到，當利用基因槍投遞不含有金粒子之裸露DNA疫苗比較利用肌肉注射疫苗之具有更好之治療效果，而推論其結果可能也與此治療方式提昇CD8+T細胞之活性相關。

The therapeutic efficacy of HER2/c-erbB-2/neu DNA immunization on mouse tumor cells expressing exogenous human or rat HER-2/neu but not on mouse tumor cells naturally expressing mouse HER-2/neu has been demonstrated. We investigated the feasibility of using N-terminal rat HER-2/neu DNA immunization on mouse tumors overexpressing endogenous HER-2/neu and enhancing the therapeutic efficacy of this vaccine by fusion to various cytokine genes, including interleukin-2(IL-2), interleukin-4(IL-4), or granulocyte–macrophage colony-stimulating factor. In a therapeutic model, N’-neu-IL-2 DNA was significantly better than N′-neu DNA vaccine and the therapeutic efficacy of DNA vaccines was correlated with tumor infiltration of CD8+ T cells. To further enhance the efficacy of DNA vaccine, we investigated the therapeutic effect of the combination of geldanamycin and N′-neu DNA vaccine against Hsp90 clients HER-2/neu protein. The efficacy of various doses of geldanamyc in N’-neu DNA vaccine was investigated in a tumor constitutively overexpressing endogenous HER-2/neu. Low-dose(2.5 �g), but not high-dose(10 �g), geldanamycin had an enhancing effect with N’-neu DNA vaccine on the inhibition of MBT-2 tumors in syngeneic C3H mice. Anti-HER-2/neu antibody titers were similar among all treated groups. Significant increased infiltrations of CD8+ T cells were observed at tumor sites. Geldanamycin sensitized tumor cells to the cytotoxic effects of lymphocytes. Furthermore, to enhance the efficacy of DNA vaccine, we also investigated the therapeutic effects of different procedures of DNA vaccine delivery. The results indicated that delivery of naked N’-neu DNA using gene gun showed better therapeutic efficacy than that using intramuscular injection. The therapeutic effect was correlated with increased CD8+ T cell activity.

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