摘要
目的评价肿瘤疫苗在结肠癌模型小鼠体内的抑制效果。方法用CpG的β-葡聚糖纳米颗粒(CpGβ-glucan nanoparticles,CNP)于体外刺激小鼠骨髓来源树突状细胞(bone marrow-derived dendritic cells,BMDCs),同时设PBS组、NP组(无CpG纳米颗粒)、Lysate组(MC38细胞裂解物)及CpG组(CpG1826),流式细胞术检测BMDCs表面标志分子的表达情况,ELISA法检测细胞培养上清中白细胞介素-6(interleukin-6,IL-6)和IL-12p40的含量。将50 mg/mL肿瘤裂解物(MC38细胞裂解物)与200 mg/mL的CNP按1∶1的体积比混合,制备成肿瘤裂解物纳米疫苗。用该疫苗经皮下免疫C57BL/6J小鼠(Vaccine组),同时设PBS组、CNP组及Lysate组,每周1次,共免疫3次,末次免疫后1 h,经小鼠右下肢皮下接种MC38细胞,2×10^(5)个/只,每3 d测量1次肿瘤体积,并绘制肿瘤生长曲线;流式细胞术分析小鼠血液中CD3^(+)CD4^(+)T及CD3^(+)CD8^(+)T细胞比例;ELISA法检测小鼠血液和脾脏中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和干扰素γ(interferonγ,IFNγ)的含量。结果CNP在体外提高BMDCs表面标志物CD11c^(+)CD80^(+)、CD11c^(+)CD86^(+)、CD11c^(+)MHC-Ⅱ^(+)的表达及IL-6和IL-12p40的分泌水平明显高于其他4组(t=4.3~46.2,P均<0.05)。与其他3组比较,Vaccine组小鼠肿瘤体积明显减小(t=2.6~3.4,P均<0.05);CD3^(+)CD8^(+)T及CD3^(+)CD8^(+)T细胞比例差异均无统计学意义(t=0.5~1.9,P均>0.05);血液中IFNγ含量明显升高(t=3.8~4.6,P均<0.05),TNF-α含量差异无统计学意义(t=0.4~2.0,P均>0.05);脾脏中IFNγ及TNF-α含量均明显升高(t=6.3~13.0,P均<0.001)。结论制备的肿瘤裂解物纳米疫苗能提高小鼠体内免疫水平,有效抑制结肠癌生长。
Objective To evaluate the inhibitory effect of tumor vaccines in colon carcinoma model mice.Methods Mouse bone marrow-derived dendritic cells(BMDCs)were stimulated by using CpGβ-glucan nanoparticles(CNP)in vitro.The BMDCs were divided into PBS group,NP group(without CpG nanoparticles),Lysate group(MC38 cell lysate)and CpG group(CpG1826),which were determined for the expression of marker molecules on the surface by flow cytometry and for the contents of interleukin-6(IL-6)and IL-12p40 in the culture supernatant by ELISA.The tumor lysate nano-vaccine was prepared by mixing 50 mg/mL tumor lysate(MC38 cell lysate)with 200 mg/mL CNP in a volume ratio of 1∶1,with which mice were subcutaneously immunized as Vaccine group.Vaccine group,PBS group,CNP group and Lysate group were immunized once a week,for three times in total.Mice were subcutaneously inoculated with MC38 cells,2×10~5 cells for each,in the right lower limb 1 h after the last immunization,and measured for tumor volume once every three days to plot the tumor growth curve.The ratios of CD3^(+)CD4^(+)T and CD3^(+)CD8^(+)T cells in the blood were analyzed by flow cytometry and the levels of tumor necrosis factor-α(TNF-α)and interferonγ(IFNγ)in the blood and spleen of mice were determined by ELISA.Results CNP effectively increased the expression of CD11c^(+)CD80^(+),CD11c^(+)CD86^(+),CD11c^(+)MHC-Ⅱ^(+)and the secretion of IL-6 and IL-12p40 in BMDCs in vitro,which were significantly higher than those in other 4 groups(t=4.3~46.2,each P<0.05).Compared with that of the other three groups,the tumor volume of mice in Vaccine group decreased significantly(t=2.6~3.4,each P<0.05);There was no significant difference in CD3^(+)CD8^(+)T and CD3^(+)CD8^(+)T cell ratios(t=0.5~1.9,each P>0.05);The content of IFNγin blood increased significantly(t=3.8~4.6,P<0.05),while that of TNF-αshowed no significant difference(t=0.4~2.0,each P>0.05);However,the contents of IFNγand TNF-αin spleen increased significantly(t=6.3~13.0,each P<0.001).Conclusion The prepared nano-vacci
作者
韩卢
梁朝远
石思伟
杨立群
邓雄威
盛望
HAN Lu;LIANG Zhao-yuan;SHI Si-wei;YANG Li-qun;DENG Xiong-wei;SHENG Wang(Faculty of Environment and Life,Beijing University of Technology,Beijing 100124,China;不详)
出处
《中国生物制品学杂志》
CAS
CSCD
北大核心
2023年第1期11-15,20,共6页
Chinese Journal of Biologicals
基金
国家自然科学基金(31770999)。
