Vruzhaj Idris, Gambirasi Marta, Busato Davide, Giacomin Aurora, Toffoli Giuseppe, Safa Amin
Experimental and Clinical Pharmacology Unit, Centro di Riferimento Oncologico di Aviano (CRO) IRCCS, 33081 Aviano, Italy.
Department of Life Sciences, University of Trieste, 34127 Trieste, Italy.
Medicina (Kaunas). 2025 Mar 30;61(4):633. doi: 10.3390/medicina61040633.
: The administration of oral vaccines offers a potential strategy for cancer immunotherapy; yet, the development of effective platforms continues to pose a difficulty. This study examines Nissle 1917 (EcN) as a microbial vector for the precise delivery of Glypican-1 (GPC1), a tumor-associated antigen significantly overexpressed in pancreatic ductal adenocarcinoma (PDAC).To evaluate the effectiveness of EcN as a vector for the delivery of GPC1 and assess its potential as an oral vaccination platform for cancer immunotherapy. : EcN was genetically modified to produce a GPC1-flagellin fusion protein (GPC1-FL) to augment antigen immunogenicity. The expression and stability of GPC1 were confirmed in modified PANC02 cells using Western blot and flow cytometry, indicating that GPC1 expression did not influence tumor cell growth. A mouse model was employed to test immunogenicity post-oral delivery, measuring systemic IgG, IL-10, IL-2, and IFN-γ levels to indicate immune activation. : Oral immunization with EcN GPC1-FL elicited a robust systemic immune response, demonstrated by markedly increased levels of IgG and IL-10. IL-2 and IFN-γ concentrations were elevated in vaccinated mice relative to controls; however, the differences lacked statistical significance. Western blot examination of fecal samples verified consistent antigen expression in the gastrointestinal tract, indicating effective bacterial colonization and antigen retention. No detrimental impacts were noted, hence substantiating the safety of this methodology. : These findings confirm EcN as a feasible and patient-friendly oral vaccination platform for cancer immunotherapy. The effective production of GPC1 in tumor cells, along with continuous antigen delivery and immune activation, underscores the promise of this approach for PDAC and other cancers. This study promotes microbial-based antigen delivery as a scalable, non-invasive substitute for traditional vaccine platforms.
口服疫苗的给药为癌症免疫治疗提供了一种潜在策略；然而，有效平台的开发仍然存在困难。本研究考察了1917年的大肠杆菌Nissle株（EcN）作为一种微生物载体，用于精确递送Glypican-1（GPC1），这是一种在胰腺导管腺癌（PDAC）中显著过表达的肿瘤相关抗原。以评估EcN作为递送GPC1的载体的有效性，并评估其作为癌症免疫治疗口服疫苗平台的潜力。：对EcN进行基因改造，以产生GPC1-鞭毛蛋白融合蛋白（GPC1-FL），以增强抗原免疫原性。使用蛋白质免疫印迹法和流式细胞术在修饰的PANC02细胞中证实了GPC1的表达和稳定性，表明GPC1的表达不影响肿瘤细胞生长。采用小鼠模型测试口服给药后的免疫原性，测量全身IgG、IL-10、IL-2和IFN-γ水平以指示免疫激活。：用EcN GPC1-FL进行口服免疫引发了强烈的全身免疫反应，表现为IgG和IL-10水平显著升高。与对照组相比，接种疫苗的小鼠体内IL-2和IFN-γ浓度升高；然而，差异缺乏统计学意义。粪便样本的蛋白质免疫印迹检查证实了胃肠道中抗原表达一致，表明细菌有效定植和抗原保留。未观察到有害影响，因此证实了该方法的安全性。：这些发现证实EcN是一种可行且对患者友好的癌症免疫治疗口服疫苗平台。GPC1在肿瘤细胞中的有效产生，以及持续的抗原递送和免疫激活，突出了这种方法对PDAC和其他癌症的前景。本研究促进了基于微生物的抗原递送作为传统疫苗平台的一种可扩展、非侵入性替代方法。
