Abstract
Helicobacter pylori (H. pylori) is a key cause of chronic inflammation and gastric cancer, and Epstein-Barr virus (EBV) also contributes to gastric tumorigenesis. However, the role of EBV co-infection in H. pylori-related cancers remains unclear because of the lack of efficient ex vivo models. Organoids, which are derived from self-renewing stem cells, closely mimic in situ tissue structure and function, making them ideal for disease modeling. Here, we developed patient-derived normal gastric organoids (NGOs) and used high-accuracy microinjection to introduce EBV and H. pylori (NCTC11637 strain). We found that the co-infection of H. pylori and EBV induced a significant structural change and upregulated the expression of TFF1, VIL1, and Lgr5 to promote cell proliferation and tissue morphogenesis. Analysis of transmission electron microscopy revealed an increase of H. pylori internal location in NGOs after co-infection. Moreover, all tested H. pylori strains isolated from patients displayed similar toxicities when co-infected with EBV. These findings provide a fundamental basis for evaluating pathogen toxicity, predicting disease progression, and advancing the clinical treatment of gastric cancer. IMPORTANCE: Helicobacter pylori (H. pylori) infection is a major contributor to chronic inflammation and the development of gastric cancer. Furthermore, Epstein-Barr virus (EBV) has been shown to play a role in the oncogenic process of gastric cancer by promoting chronic inflammation and increasing tissue damage. However, the mechanism by which co-infection contributes to gastric carcinogenesis remains unclear. In this study, we used patient-derived gastric organoids as a model to establish EBV-H. pylori co-infection using microinjection technology and found that co-infection causes significant structural changes and promotes cell proliferation. This model will not only contribute to a better understanding of the pathogenesis of gastric cancer but will also be important for drug efficacy evaluation and the development of new therapeutic approaches.