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目的制备纳米乳剂包裹基因工程MAGE1(melanoma antigen1)抗原,研究其生物学特性及抗肿瘤免疫效应。方法采用界面聚合法,制备包裹MAGE1抗原的纳米乳剂(NEMAGE1),测量粒径、包裹率、载药量。检测乳剂包裹对小鼠DC摄取抗原能力的影响。用NEMAGE1免疫小鼠,运用酶联免疫斑点法(ELISPOT)和细胞毒性杀伤实验(LDH)检测NEMAGE1激活机体细胞免疫反应的状况,并观察纳米蛋白疫苗对荷瘤小鼠的治疗作用。结果成功制备粒径为(15±5)nm的纳米乳剂,包裹率为91%,载药量0.091gPL,4℃放置6个月后性质稳定。DC摄取NEMAGE1的能力明显强于对游离蛋白抗原的摄取。ELISPOT和细胞毒性杀伤实验显示,NEMAGE1可以诱导机体产生针对MAGE1的CTL,特异性杀伤表达MAGE1的肿瘤细胞。荷瘤小鼠的治疗实验显示,NEMAGE1对表达MAGE1的肿瘤有明显疗效,其效果优于单独使用游离MAGE1蛋白。结论纳米乳剂具有较高包裹率和稳定性。纳米乳剂包裹的肿瘤特异性抗原可以有效激活机体产生抗肿瘤免疫效应,是具有临床应用前景的新型抗肿瘤疫苗。
Objective To prepare nanoemulsion encapsulated genetically engineered MAGE1 (melanoma antigen1) antigen and study its biological characteristics and anti-tumor immune effect. Methods The interfacial polymerization method was used to prepare nano-emulsion (NEMAGE1) wrapped with MAGE1 antigen. The particle size, encapsulation efficiency and drug loading were measured. The effect of emulsion package on DC uptake of antigen in mice was examined. The mice were immunized with NEMAGE1 and the cellular immune response by NEMAGE1 was detected by ELISPOT and LDH. The therapeutic effect of nano-protein vaccine on tumor-bearing mice was also observed. Results Nanoemulsion with particle size of (15 ± 5) nm was prepared successfully. The encapsulation efficiency was 91% and the drug loading was 0.091gPL. After 4 months at 6 ℃, the nanoemulsion was stable. The ability of DC to ingest NEMAGE1 is significantly stronger than the uptake of free protein antigens. ELISPOT and cytotoxicity experiments showed that NEMAGE1 can induce the body to produce CTL against MAGE1 and specifically kill MAGE1-expressing tumor cells. Treatment experiments in tumor-bearing mice showed that NEMAGE1 has a significant effect on tumors expressing MAGE1, and its effect is superior to that of using free MAGE1 protein alone. Conclusion Nanoemulsion has higher parcel rate and stability. Nano-emulsion-coated tumor-specific antigen can effectively activate the body to produce anti-tumor immune effect, is a new anti-tumor vaccine with clinical application prospects.