Drolia Rishi, Bhunia Arun K
Cellular and Molecular Microbiology Laboratory, Department of Biological Science, Old Dominion University, Norfolk, VA, USA.
Center for Bioelectronics, Old Dominion University, Norfolk, VA, USA.
Methods Mol Biol. 2025;2942:1-21. doi: 10.1007/978-1-0716-4627-4_1.
The gastrointestinal tract is a crucial battleground in microbial pathogenesis, where pathogens must adhere to, invade, and translocate through intestinal epithelial cells to establish infection and cause disease. Investigating these processes is critical for understanding the mechanisms of infection and host defense. In vitro assays using intestinal cell models enable precise analysis of microbial adhesion, invasion, and translocation at the cellular level. In contrast, in vivo, rodent models provide valuable insights into the complex interactions between pathogens, host tissues, and immune responses. Advanced imaging techniques, such as confocal microscopy, offer high-resolution visualization of host-pathogen interactions, allowing researchers to observe pathogen-induced alterations to the intestinal epithelial barrier and study microbial dissemination. By combining functional assays with cutting-edge imaging technologies, researchers can uncover critical mechanisms that drive microbial pathogenesis, revealing how pathogens breach intestinal defenses and spread systemically. This integrated approach deepens our understanding of infection dynamics and facilitates the development of new therapeutic strategies to combat gastrointestinal infections. Here, we describe a comprehensive approach to studying microbial pathogenesis by integrating in vitro and in vivo assays of intestinal cell adhesion, invasion, and translocation alongside advanced imaging techniques, such as confocal microscopy using immunostaining and fluorescence in situ hybridization (FISH) in tissue sections, to elucidate host-pathogen interactions and infection dynamics at the cellular level.
胃肠道是微生物致病过程中的关键战场，病原体必须在此处黏附、侵入并穿过肠道上皮细胞进行移位，以建立感染并引发疾病。研究这些过程对于理解感染机制和宿主防御至关重要。使用肠道细胞模型进行的体外试验能够在细胞水平上精确分析微生物的黏附、侵入和移位。相比之下，在体内，啮齿动物模型为病原体、宿主组织和免疫反应之间的复杂相互作用提供了有价值的见解。先进的成像技术，如共聚焦显微镜，可对宿主-病原体相互作用进行高分辨率可视化，使研究人员能够观察病原体引起的肠道上皮屏障改变，并研究微生物的传播。通过将功能试验与前沿成像技术相结合，研究人员可以揭示驱动微生物致病的关键机制，揭示病原体如何突破肠道防御并全身扩散。这种综合方法加深了我们对感染动态的理解，并促进了对抗胃肠道感染的新治疗策略的开发。在此，我们描述一种综合体外和体内肠道细胞黏附、侵入和移位试验以及先进成像技术（如在组织切片中使用免疫染色和荧光原位杂交（FISH）的共聚焦显微镜）来研究微生物致病机制的全面方法，以阐明细胞水平上的宿主-病原体相互作用和感染动态。
