纳米肿瘤疫苗抗小鼠黑色素瘤的免疫效果

Immune effects of nano-tumor vaccine against melanoma in mice

目的 研究基于纳米复合佐剂[包载CpG的纳米颗粒(CpG-coated nanoparticles,CNP)]及黑色素瘤细胞裂解物抗原的纳米肿瘤疫苗的抗黑色素瘤免疫效果。方法 检测CNP及黑色素瘤细胞裂解物抗原对小鼠骨髓来源树突状细胞(bone marrow-derived dendritic cells,BMDCs)的免疫调控作用及其对细胞因子IL-6和IL-12表达分泌的调控作用。接种黑色素瘤细胞的小鼠成瘤后,选择肿瘤体积相近的40只雌性C57BL/6N小鼠随机分为4组:对照(PBS)、佐剂(CNP)、裂解液(Lysate)和疫苗(CNP+Lysate)组,按皮下接种方式给药,1周1次,给药3周;每3 d记录小鼠肿瘤大小并绘制肿瘤生长曲线;收集小鼠外周血检测IFN-γ和TNF-α含量;免疫组化法检测CD8^(+)T淋巴细胞在肿瘤组织中的浸润情况。结果 与PBS、CpG和肿瘤裂解物抗原组相比,纳米疫苗佐剂CNP能有效刺激BMDCs成熟,促进IL-6和IL-12分泌;接种纳米肿瘤疫苗显示出较好的体内抗肿瘤活性,疫苗组小鼠肿瘤体积明显减小,其血清中IFNγ和TNF-α分泌水平显著高于其他组;疫苗组小鼠肿瘤中CD8^(+)T淋巴细胞浸润情况也明显优于其他组。结论 纳米肿瘤疫苗能有效激活BMDCs并高表达免疫因子,也能有效抑制肿瘤生长,显示出良好的应用潜质。

Objective To investigate the anti-melanoma immune effects of nano-tumor vaccine based on nano-adjuvant[CpG-coated nanoparticles(CNP)] and melanoma cell lysate antigen.Methods The immunoregulatory effects of CNP and melanoma cell lysate antigens on bone marrow-derived dendritic cells(BMDCs) and the regulatory effects on expression and secretion of cytokines IL-6 and IL-12 were investigated.After the mice inoculated with melanoma cells formed tumor,40C57BL/6N fermale mice with similar size of tumor were randomly divided into 4 groups:control(PBS) group,adjuvant(CNP) group,lysate(Lysate) group and vaccine(CNP+Lysate) group,which were administered subcutaneously once a week for 3 weeks.The tumor size of mice was recorded every 3 d and the tumor growth curve was drawn.The peripheral blood of mice was collected to detect the contents of IFN_γ and TNF_α,and immunohistochemical method was used to detect the infiltration of CD8^(+)T lymphocytes in tumor tissues.Results Compared with PBS,CpG and tumor lysate antigen groups,nano-vaccine adjuvant CNP effectively stimulated BMDCs maturation and promoted IL-6 and IL-12 secretion;Nanotumor vaccine showed good anti-tumor activity in vivo,the tumor size of mice in vaccine group decreased significantly,and the secretion levels of IFN_γ and TNF-α in serum were significantly higher than those in other groups;The infiltration of CD8^(+)T lymphocytes in tumor tissues of mice in vaccine group was also significantly better than that in other groups.Conclusion Nano-tumor vaccine effectively activated BMDCs,highly expressed immune factors,and also effectively inhibited tumor growth,showing good application potential.