载乳腺癌裂解蛋白纳米粒子的制备及其体外免疫效应的初步评估

Preparation of Breast Cancer Cell Lysates-loaded Nanoparticle and Preliminary Evaluation of Its Immunity in vitro

[目的]使用聚乳酸聚乙醇酸共聚物(PLGA)制备纳米颗粒,用来包载MCF7乳腺癌细胞的裂解蛋白,并初步评估其在乳腺癌免疫治疗中的作用。[方法]用二次乳化法制备载MCF7裂解蛋白的纳米粒子,对其包封效率、粒径、形态和Zeta电位进行测定,优化制备参数以提高包封效率;用该纳米粒子刺激人外周血单个核细胞,检测纳米粒子的体外抗肿瘤免疫效应。[结果]制备的载MCF7裂解蛋白纳米颗粒呈球形,尺寸较均匀,平均粒径75nm,表面带负电荷,对MCF7裂解蛋白的包裹率可达85%,并能显著提高人淋巴细胞对乳腺癌细胞的体外杀伤能力,约是对照组的3倍(P<0.05)。[结论]该制备方法所得载MCF7裂解蛋白纳米粒理化性质优良,包裹率高,在体外实验中可提高MCF7裂解蛋白的免疫原性,在提升抗肿瘤免疫效应方面具有应用潜能。

[Purpose] To prepare nanoparticle for encapsulating MCF7 breast cancer cell lysate protein （MCFTp） with poly （D, L-lactide co-glycolide） copolymer （PLGA）, and to eval- uate its role in immunotherapy for breast cancer. [Methods] The double emulsion method was employed for preparation of nanoparticles （NP） encapsulating the MCF7 breast cancer cell lysate protein. ]-he morphology, size, zeta potential, and encapsulation efficiency of the NP were assayed and optimized. The NP was utilized to simulate the peripheral blood mononuclear cells （PBMC）, and the anti-tumor immune reaction of NP was detected in vitro. [ Results] The obtained NPs were negatively charged, spherical in shape, relatively uniform in size, with a mean diameter of 75nm and a protein encapsulation rate of 85%.The NP-stimulated PMBC generated a significantly higher anticancer cytotoxicity in vitro, with 3 times compared to the control group（P〈0.05）. [Conclusion] The NPs prepared are of good physicochemical properties and can encapsulate protein antigen with high efficiency. Moreover, the NPs significantly enhance the immunogenicity of the MCF7 tumor lysate protein, suggesting that nanotechnology may have potential applications in immunotherapy against cancer.