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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

31卷

期数:

2025年03期

页码:

189-195

栏目:

肿瘤疾病专题

出版日期:

2025-05-20

文章信息/Info

Title:

Oral administration of tumor antigen-loaded bacterial outer membrane vesicle vaccines effectively inhibits hepatocellular carcinoma

文章编号:

1006-8147(2025)03-0189-07

作者:

张亚丽; 张杨; 钟彦至; 左冰峰; 尹海芳

（天津医科大学医学技术学院，天津 300070）

Author(s):

ZHANG Yali; ZHANG Yang; ZHONG Yanzhi; ZUO Bingfeng; YIN Haifang

（School of Medical Technology，Tianjin Medical University，Tianjin 300070，China）

关键词:

肝细胞癌; 细菌外膜囊泡; 细胞穿膜肽; 口服疫苗

Keywords:

-

分类号:

Q291

DOI:

10.20135/j.issn.1006-8147.2025.03.0189

文献标志码:

A

摘要:

目的：利用负载肿瘤抗原的细菌外膜囊泡（OMVs）作为口服疫苗，以探究其抑制肝细胞癌的效果。方法：首先，利用超速离心法收集益生菌EcN 1917来源的OMVs，并通过电镜和DLS表征其形态及粒径；其次，利用考染和流式检测TAT介导蛋白负载于OMVs内部的效率及抗胃蛋白酶消化的能力；利用transwell实验模拟肠上皮屏障，评估OMVs透过肠上皮屏障诱导树突状细胞（DC）活化的能力；最后，在表达模式识别抗原OVA的小鼠肝细胞癌皮下瘤Hepa1-6-OVA模型上，评估经口服OMVOVA疫苗诱导的肿瘤特异性免疫应答效应。结果：OMV为球形脂质双层纳米囊泡，平均直径为37.8 nm；流式检测结果显示：TAT细胞穿膜肽可介导GFP荧光报告蛋白在OMVs内部高效负载并可保护GFP蛋白不被消化降解；体外模拟肠上皮实验结果证明：OMVs可穿过肠上皮屏障并激活DC，上调共刺激分子CD80/CD86表达（t=5.571、57.64，均P<0.01）；在表达OVA抗原的Hepa1-6肝细胞癌皮下瘤小鼠模型中，口服OMVOVA可有效抑制肿瘤生长，肿瘤体积（t=4.288，P<0.01）和肿瘤质量（t=5.812，P<0.01）均显著降低，肠系膜淋巴结中DC活化（t=10.08，P<0.01）及对抗原肽的提呈（t=5.853，P<0.01）均显著增高，显著激活抗原特异性T细胞数量（t=7.863，P<0.01）。结论：口服内部负载肿瘤抗原肽新型OMV疫苗，可有效抑制小鼠肝细胞癌皮下瘤生长，激活黏膜免疫诱导抗原特异性免疫应答。

Abstract:

Objective： To investigate the inhibitory effect of bacterial outer membrane vesicles （OMVs） on hepatocellular carcinoma using OMVs loaded with tumor antigens as oral vaccines. Methods：OMVs derived from probiotic EcN 1917 were collected by ultracentrifugation，and their morphology and particle size were characterized by transmission electron microscopy and DLS. The loading efficiency of TAT mediated protein loaded into OMVs and the ability to resist gastric protease digestion were examined by Coomassie blue staining and flow cytometry analysis. The ability of OMVs penetrating the intestinal epithelial barrier and inducing dendritic cell （DC） acti-vation was measured by transwell test. The antitumor specific immune responses induced by oral OMVOVA vaccine were evaluated in subcutaneous HCC mouse models Hepa1-6-OVA bearing OVA-overexpressing antigens. Results：The OMVs were spherical lipid bilayer nanovesicles with an average diameter of 37.8 nm. The results of flow cytometry showed that TAT cell-penetrating peptide could efficiently load GFP fluorescent reporter protein in OMVs and protect GFP protein from digestion and degradation. The results of in vitro intestinal epithelial experiment showed that OMVs could cross the intestinal epithelial barrier and activate DC，and up-regulate the expression of costimulatory molecules CD80/CD86 （t=5.571，57.64，both P<0.01）. Oral administration of OMVOVA significantly inhibited tumor growth in OVA-expressing subcutaneous Hepa1-6 hepatocellular carcinoma mice，with significantly reduced tumor volume （t=4.288，P<0.01） and tumor weight （t=5.812，P<0.01）. DC activation （t=10.08，P<0.01） and antigen peptide presentation （t=5.853，P<0.01） in mesenteric lymph nodes were significantly increased，and the number of antigen specific T cells was significantly activated （t=7.863，P<0.01）. Conclusion：The oral administration of the novel OMV vaccine loaded with tumor antigen pep

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备注/Memo

备注/Memo:

基金项目:国家重点研发计划（2301-2JC280）
作者简介:张亚丽（2000-），女，硕士在读，研究方向：药物靶向递送；
通信作者：尹海芳；E-mail：haifangyin@tmu.edu.cn。

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更新日期/Last Update: 2025-06-01