中文摘要：

目的 观察粒细胞-巨噬细胞集落刺激因子（granulocyte-macrophage colony stimulating factor,GM-CSF）作为II型登革病毒E蛋白免疫佐剂的保护效果,评估GM-CSF作为蛋白疫苗免疫佐剂的可行性。方法 提取GM-CSF质粒（pCAG-GM）、将表达II型登革病毒E蛋白的重组质粒（E/pGEX-6P-1）进行诱导,表达产物用亲和层析纯化。将BALB/c小鼠随机分为E蛋白组、佐剂组（E蛋白＋GM-CSF）和对照组进行免疫,采用酶联免疫吸附试验（ELISA）检测血清抗体终点效价;采用蚀斑减少中和试验（PRNT）检测II型登革病毒中和抗体水平;酶联免疫斑点试验（ELISPOT）检测小鼠免疫后细胞因子的水平;用攻毒试验观察各组小鼠保护率。结果 E蛋白组和佐剂组小鼠血清中抗体终点效价、中和抗体水平均在免疫后有一定的升高,差异无统计学意义（P〉0.05）;佐剂组细胞因子（IFN-γ和IL-10）水平较E蛋白组均有明显上升,差异具有统计学意义（P〈0.05）;但攻毒试验显示,佐剂组小鼠全部死亡,生存率为0,而E蛋白组的小鼠保护率为33%。结论 GM-CSF免疫佐剂可抑制II型登革病毒E蛋白的免疫保护作用,其作为蛋白疫苗免疫佐剂仍需慎重。

英文摘要：

Objective To observe and evaluate the effect of GM-CSF as a potential adjuvant in immunization with type 2 dengue virus （DV2 ） E protein. Methods The GM-CSF plasmid was extracted and recombinant E protein was induced, the expressed recombinant E protein was purified by using affinity chromatography. The experimental mice were divided into three groups including E protein group, adjuvant group （E protein＋ GM-CSF） and control group. ELISA and PRNT were used in detection of the IgG levels for anti-DV2 and neutralizing antibody levels in mice sera of each group, respectively. ELISPOT was used to detect the splenocyte-secreted cytokines. Challenge experiment was carried out in detection of the protection rate in each tested group. Results The level of total IgG and neutralizing antibody were increased in both E pro- tein and adjuvant groups, but there was no statistical meaning between two groups （ P〉0.05 ）. ELISPOT result showed that adjuvant immunization caused an increase in levels of IFN-y and IL-10 in compared to the E protein group, and there was a statistical difference between two groups（P〈0.05） , there was no mouse survival in adjuvant group while 33% survival rate was observed in E protein group. Conclusion GM-CSF could inhibit the protection induced by DV2 E protein. Thus GM-CSF should be used and evaluated with caution as long as it was used as an adjuvant by the DV protein vaccine.