肿瘤疫苗TCL/CpG@GNP的构建及评价

doi:10.11958/20251525

天津医药 ›› 2025, Vol. 53 ›› Issue (10): 1021-1026.doi: 10.11958/20251525

肿瘤疫苗TCL/CpG@GNP的构建及评价

陈敏纯(), 薛润青, 赵新, 张萌, 叶丹, 张婧一, 郑洁^△()

西北大学附属医院•西安市第三医院药剂科（邮编 710018）

收稿日期:2025-05-10 修回日期:2025-06-09 出版日期:2025-10-15 发布日期:2025-10-12
通讯作者: ^△E-mail：44682408@qq.com
作者简介:陈敏纯（1982），女，副主任药师，主要从事医院药学方面研究。E-mail：minchun.chen@163.com
基金资助:
西安市卫生健康委员会项目(2023qn05);2022白求恩·求索-药学科研能力建设项目

Development and evaluation of the tumor vaccine TCL/CpG@GNP

CHEN Minchun(), XUE Runqing, ZHAO Xin, ZHANG Meng, YE Dan, ZHANG Jingyi, ZHENG Jie^△()

Department of Pharmacy, Xi’an No.3 Hospital, the Affiliated Hospital of Northwest University, Xi’an 710018, China

Received:2025-05-10 Revised:2025-06-09 Published:2025-10-15 Online:2025-10-12
Contact: ^△E-mail：44682408@qq.com

陈敏纯, 薛润青, 赵新, 张萌, 叶丹, 张婧一, 郑洁. 肿瘤疫苗TCL/CpG@GNP的构建及评价[J]. 天津医药, 2025, 53(10): 1021-1026.

CHEN Minchun, XUE Runqing, ZHAO Xin, ZHANG Meng, YE Dan, ZHANG Jingyi, ZHENG Jie. Development and evaluation of the tumor vaccine TCL/CpG@GNP[J]. Tianjin Medical Journal, 2025, 53(10): 1021-1026.

摘要/Abstract

摘要：

目的构建含有广谱新抗原的肿瘤细胞裂解物（TCL）和CpG佐剂的共聚物胶束肿瘤疫苗，递送至淋巴结树突状细胞。方法基于一种新型聚合物9-氟烯基甲氧基碳酸酯-聚乙二醇甘油胆酸通过π-π堆积方式包裹TCL和CpG构建一种高密度聚乙二醇修饰的甘油胆酸的微胶束TCL/CpG@GNP。考察疫苗TCL/CpG@GNP的载药率和包封率，粒径、多分散指数（PDI）、电位、形态、细胞安全性，疫苗稳定性，细胞摄取能力，骨髓来源的树突状细胞（BMDC）的免疫刺激作用相关指标及体内抗肿瘤效应评估。结果 TCL/CpG@GNP疫苗TCL的载药量为6.26%，包封率为37.59%；CpG的载药率为7.05%，包封率为56.86%。粒径为（139.26±27.23）nm，PDI为0.249±0.015，具有良好的分散性能，电位为（-21.23±0.36）mV。TCL/CpG@GNP具有良好的稳定性、细胞安全性及摄取能力，能促进BMDC激活及成熟。在荷瘤小鼠模型中，TCL/CpG@GNP可抑制肿瘤生长，促进外周血T淋巴细胞比例增高及提高脾脏γ干扰素水平。结论构建的肿瘤疫苗TCL/CpG@GNP能够有效激活BMDCs，并在小鼠肺癌模型中诱导强效抗肿瘤免疫记忆。

关键词: 癌症疫苗, 肿瘤细胞裂解物, CpG佐剂, 免疫治疗

Abstract:

Objective To develop a tumor vaccine containing broad-spectrum neoantigen tumor cell lysate (TCL) and CpG adjuvant, and to effectively deliver it to lymph node dendritic cells. Methods A novel polymer, 9-fluorenylmethoxycarbonyl-polyethylene glycol-glycocholic acid (Fmoc-PEG-GCA), was employed to encapsulate the TCL and CpG through π-π stacking, resulting in high-density polyethylene glycol-modified glycocholic acid-decorated micelles TCL/CpG@GNP. The vaccine's drug loading, encapsulation efficiency, particle size, polydispersity index (PDI), zeta potential, morphology, stability, cellular safety, uptake capability, immune stimulation effects on bone marrow-derived dendritic cells (BMDCs) and in vivo anti-tumor efficacy were evaluated. Results The vaccine TCL and CpG demonstrated a drug loading capacity of 6.26% and the encapsulation rate was 37.59%. The drug loading capacity of CpG was 7.05%, and the encapsulation efficiency was 56.86%. The particle size measured (139.26±27.23) nm, with a PDI of 0.249±0.015, indicating favorable dispersion properties. The zeta potential was recorded at (-21.23±0.36) mV. The TCL/CpG@GNP vaccine demonstrated good stability, cell safety and uptake ability, and can promote the activation and maturation of BMDCs. In tumor-bearing mouse models, TCL/CpG@GNP inhibited tumor growth, increased the proportion of T lymphocytes in peripheral blood, and elevated IFN-γ levels in spleen. Conclusion The TCL/CpG@GNP tumor vaccine can effectively activate BMDCs and induce strong anti-tumor immune memory in a mouse lung cancer model.

Key words: cancer vaccines, tumor cell lysates, CpG adjuvant, immunotherapy

中图分类号:

R186.9，R730

图/表 9

图1 TCL/CpG@GNP化学结构表征

Fig.1 Chemical structure characterization of TCL/CpG@GNP

图2 TCL/CpG@GNP的电镜扫描图像（×200）

Fig.2 TEM images of TCL/CpG@GNP (×200)

图3 TCL/CpG@GNP在胃肠道的稳定性 A：TCL/CpG@GNP与人工胃液共孵育2 h的粒径变化；B：TCL/CpG@GNP与人工肠液共孵育6 h的粒径变化，a与0 h比较，b与1 h比较，c与2 h比较，d与4 h比较，P<0.05。

Fig.3 The stability of TCL/CpG@GNP in gastrointestinal tract

图4 TCL/CpG@GNP的细胞摄取能力（×200）

Fig.4 Cellular uptake ability of TCL/CpG@GNP (×200)

图5 TCL/CpG@GNP对BMDC免疫刺激成熟情况 A—C：流式细胞术分析BMDC成熟度；D：流式结果统计图，a与PBS组比较，b与TCL+CpG+GNP组比较，P<0.05。

Fig.5 Immune-stimulatory effects of TCL/CpG@GNP

表1 各组BMDC培养上清液中IL-6和IL-12 p70的含量比较（n=6，ng/L，$\bar{x}±s$）

Tab.1 Comparison of IL-6 and IL-12 p70 levels in the culture supernatants of BMDCs between the three groups

组别	IL-6	IL-12 p70
PBS组	18.22±3.30	23.45±11.17
TCL+CpG+GNP组	21.55±3.22	34.51±7.60
TCL/CpG@GNP组	35.96±3.83^ab	380.72±25.70^ab
F	22.296^＊	440.623^＊＊

图6 3组小鼠的肿瘤体积增长曲线 n=6，F2 d=5.302，P<0.05；F4 d=3.650，P>0.05；F6 d=0.634，P>0.05；F8 d=3.364，P>0.05；F11 d=59.283，P<0.01；a与PBS组比较，b与TCL+CpG+GNP组比较，P<0.05。

Fig.6 Tumor growth curves of the three groups of mice

图7 TCL/CpG@GNP刺激小鼠外周血淋巴细胞表达 A—C：流式细胞术分析BMDC成熟度；D：流式结果统计图，a与PBS组比较，b与TCL+CpG+GNP组比较，P<0.05。

Fig.7 The expression in mouse peripheral blood lymphocytes stimulated by TCL/CpG@GNP

图8 TCL/CpG@GNP刺激小鼠脾脏淋巴细胞表达 A—C：流式细胞仪分析小鼠脾脏中的IFN-γ表达；D：小鼠脾脏中IFN-γ表达的统计图，a与PBS组比较，b与TCL+CpG+GNP组比较，P<0.05。

Fig.8 The expression of mouse spleen lymphocytes stimulated by TCL/CpG@GNP

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肿瘤疫苗TCL/CpG@GNP的构建及评价

Development and evaluation of the tumor vaccine TCL/CpG@GNP

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