论文部分内容阅读
用纳米微粒包装肿瘤抗原,并探讨其诱导产生CTL的免疫学特性和杀瘤活性。采用复乳溶剂蒸发法制备包含有恶性黑色素瘤抗原肽Mart-1-27-35纳米微粒;扫描电镜观察所制得纳米颗粒的形态。用NP-Mart-1-27-35诱导抗原特异性CTL并用ELISPOT法和LDH法检测其特性和杀瘤活性。结果:电镜观察所制得的纳米颗粒为圆形,平均直径为(208.52±12.43)nm。HPLC分析纳米-抗原肽的抗原载入量为6.72%±0.28%,平均包封率为91.23%±3.56%;ELISPOT检测表明纳米微粒包装的抗原肽能引起较强的免疫反应,产生分泌IFN-γ的细胞斑点数分别为(476.37±29.43)/2×104,显著高于单纯使用抗原肽时所产生的斑点数;LDH检测显示与单纯使用抗原肽相比,纳米-抗原肽诱导的CTL对Mart-1+细胞的杀伤能力显著提高。纳米微粒包装的抗原肽能够显著增强并延长DC对抗原的提呈作用;其诱导产生的CTL能以MHC I限制的方式特异识别和杀伤表达相应抗原的肿瘤细胞。
Tumor antigens were packaged with nanoparticles, and the immunological properties and cytotoxic activity of CTLs induced by CTLs were investigated. Mart-1-27-35 nanoparticles containing the malignant melanoma antigen peptide were prepared by double emulsion solvent evaporation method. The morphology of the prepared nanoparticles was observed by scanning electron microscopy. Antigen-specific CTLs were induced with NP-Mart-1-27-35 and their characteristics and cytotoxic activity were tested by ELISPOT and LDH assays. Results: The nanoparticles obtained by electron microscopy were round with average diameter of (208.52 ± 12.43) nm. The results of ELISPOT assay showed that the antigenic peptides encapsulated by nanoparticles could induce a strong immune response and produce IFN- (476.37 ± 29.43) / 2 × 104, respectively, which was significantly higher than that of the control group when using antigenic peptide alone. LDH showed that the CTL induced by nano-antigen peptide Mart-1 + cells have significantly increased lethality. Nanoparticle-encapsulated antigenic peptides can significantly enhance and prolong the antigen presenting function of DCs; CTLs induced by them can specifically recognize and kill tumor cells expressing the corresponding antigens in a MHC I-restricted manner.