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目的:制备包裹有MAGE1-Hsp70和SEA的复合抗原的纳米乳剂疫苗NE(MHS)(nanoemulsion-encapsulated MAGE1-Hsp70/SEA),评价其抗肿瘤免疫学特性。方法:采用磁力超声法制备纳米疫苗NE(MHS),评价其粒径、包裹率及稳定性;NE(MHS)免疫动物,IFN-γ ELISPOT和LDH杀伤实验检测疫苗激活机体特异性细胞免疫反应情况;肿瘤攻击实验检测纳米乳剂疫苗的抗肿瘤效应。结果:(1)成功制备出粒径为(20±5)nm的NE(MHS),其药物包裹率为87%,于4℃放置6个月后或3000r/min10min离心后无分层;(2)与其他组相比,NE(MHS)组小鼠脾淋巴细胞中分泌IFN-γ的T细胞数量明显增多(P<0.05),CTL对B16-MAGE-1的特异性杀伤作用明显增强;NE(MHS)组B16-MAGE-1肿瘤成瘤时间长、成瘤率低。结论:NE(MHS)纳米乳剂疫苗具有良好的理化性质,能够刺激机体产生强烈的MAGE-1特异性的细胞免疫,能有效预防表达MAGE-1的肿瘤细胞攻击,是一种很有希望的新型抗肿瘤疫苗。
OBJECTIVE: To prepare nanoemulsion-encapsulated MAGE1-Hsp70 / SEA complexed with complex antigens of MAGE1-Hsp70 and SEA, and to evaluate its anti-tumor immunological properties. Methods: The nano-vaccine NE (MHS) was prepared by magnetic-ultrasonic method to evaluate the particle size, encapsulation efficiency and stability. The immune response of NE (MHS), IFN-γ ELISPOT and LDH ; Tumor Assay Experiment to Detect Antitumor Effect of Nanoemulsion Vaccine. Results: (1) NE (MHS) with particle size of (20 ± 5) nm was successfully prepared, and the drug entrapment efficiency was 87%. After being placed at 4 ℃ for 6 months or centrifugation at 3000r / min for 10min, 2) Compared with other groups, the number of T lymphocyte secreting IFN-γ in NE (MHS) group increased significantly (P <0.05), and the specific killing effect of CTL on B16-MAGE-1 was significantly increased. NE (MHS) group B16-MAGE-1 tumor formation time, tumor formation rate is low. CONCLUSION: The NE (MHS) nanoemulsion vaccine has good physico-chemical properties and can stimulate the body to produce strong MAGE-1-specific cellular immunity, which can effectively prevent the tumor cells from attacking MAGE-1. It is a promising new type Anti-tumor vaccine.