近年來，高致病性禽流感病毒H5N1對家禽和人類造成嚴重的威脅；從病毒的構造看來，HA是流感病毒最主要的膜蛋白，因此經常作為疫苗開發的首要目標；帶有HA的細菌質體DNA已被用來作為DNA疫苗，但其免疫效果並不理想；因此大部分的DNA疫苗皆須借助佐劑提高免疫能力。由於CD40L、Flt3L和GM-CSF皆被視為調節免疫反應的重要因子，本實驗便以分別帶有CD40L、Flt3L和GM-CSF的質體DNA當作分子佐劑，期望能增強H5N1流感之DNA疫苗的免疫作用。第一次免疫試驗的結果發現，在肌肉注射三劑DNA疫苗後，同時攝入帶有H5HA和GM-CSF質體DNA的小鼠有較好的體液免疫反應。然而利用DNA prime和VLP boost的免疫試驗結果卻顯示，雖然GM-CSF可以明顯提高H5HA-specific total IgG抗體的含量，同時注射帶有H5HA和CD40L的質體DNA，對於血清中的HI和中和性抗體反而產生較好的促進效果。最後再以DNA prime和rH5HA protein boost進行一次免疫試驗，結果顯示CD40L和GM-CSF皆可促進血清中功能性抗體的產生。總而言之，雖然實驗結果並不顯著，但利用帶有CD40L和GM-CSF的細菌質體DNA進行免疫，確實能稍微提高血清中anti-H5HA的抗體含量，因此這兩種細胞激素皆具備有作為H5N1禽流感病毒DNA疫苗之分子佐劑的能力。

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