纳米复合佐剂的抗肿瘤免疫调控作用及效果研究

Anti-Tumor Immunologic Modulation and Efficacy of Nano-Complex Adjuvant

目的:探究纳米复合佐剂在肿瘤内的免疫调控作用及抗肿瘤效果。方法:制备包载CpG的β葡聚糖纳米颗粒(CNP),体外培养小鼠骨髓来源的树突细胞(BMDC),分5组进行体外免疫处理,分别为PBS对照组、β葡聚糖组、β葡聚糖纳米载体(NP)组、CpG组和CNP组,利用流式仪分析BMDC成熟情况。将体外培养的黑色素瘤B16F10细胞接种于C57BL/6N小鼠右后肢皮下,建立肿瘤模型,第6 d挑选肿瘤大小相对一致的24只小鼠均分为4组,即PBS组、NP组、CpG组、CNP组进行治疗,后颈部皮下注射给药,共给药4次,每次间隔3 d,期间隔天测量肿瘤体积,最后一次给药后第3 d眼眶取血,测定血浆中γ干扰素(IFN-γ)浓度,处死小鼠后分离肿瘤及脾脏,流式细胞术检测瘤内浸润性T淋巴细胞比例及脾脏内T细胞比例。结果:CNP佐剂处理后BMDC成熟比例约为CpG佐剂组的2.8倍。在小鼠体内接种肿瘤15 d后,CNP治疗组肿瘤体积较其他组均减小,血浆中IFN-γ含量显著高于其他实验组,约为CpG组的1.5倍,肿瘤浸润性T淋巴细胞与其他组相比也有明显升高。结论:构建的葡聚糖-CpG纳米复合佐剂能够有效激活BMDC,改善肿瘤微环境,具有良好的体内抗肿瘤效果。

Objective:To investigate the immunomodulatory and antitumor effects ofβ-glucan-based nano-adjuvant in tumor.Methods:Adjuvant CpG was encapsulated inβ-glucan-based nanoparticles(CNP)to generate nanoadjuvant.The dendritic cells derived from bone marrow(BMDC)of mice were treated by PBS(control group),β-glucan,β-glucan-based nanoparticles(NP),free CpG and CNP.BMDC maturation was measured by flow cytometry.Melanoma B16 F10 cells were inoculated subcutaneously on the right hind limb of C57 BL/6 N mice to establish tumor model.24 mice with the similar tumor size were selected at the 6 th day.The mice were divided into PBS group,NP group,CPG group and CNP group for treatment,which were injected subcutaneously at the back of the neck every three day for 4 times.The tumor volume was measured every two days.The blood was collected three days after the last administration.The concentration of IFN-γin the plasma was measured.The tumor and spleen were collected at the end point of the experiments.The proportion of infiltrative T lymphocytes in tumor and T cells in the spleen were measured by flow cytometry.Results:The mature ratio of BMDC treated with CNP was2.8 times that of CpG group.After 15 days of tumor inoculation in mice,the tumor volume in the CNP treatment group decreased compared with other groups,the IFN-γcontent in plasma was significantly higher than other experimental groups,about 1.5 times of that in CpG group,and the tumor infiltrating T-lymphocytes were also significantly higher than other groups.Conclusion:The CNP nano-adjuvant can effectively activate BMDC,improve tumor microenvironment,and have good antitumor effect in vivo.