My work focuses on elucidating the mechanisms of infectious and inflammatory diseases while advancing biotechnological innovation and driving the development of non-animal methods in infection research. We leverage complex in vitro systems to investigate the plasticity of myeloid immune cells and their role in shaping immune responses in the context of human microbiome–host interactions. We therefore integrate 3D cell culture, tissue engineering, microfluidics, and advanced microscopy with AI-driven image analysis to generate high-content, mechanistic data. Using these technologies, we recapitulate key aspects of human tissue physiology and host–pathogen interfaces. These models provide a human-relevant platform for dissecting infectious and inflammatory processes without relying on animal experimentation. The overarching aim of this work is to gain translationally relevant insights into host–pathogen and inflammatory processes and to support the development of innovative strategies for studying infectious and inflammatory diseases. At the same time, our research contributes to the broader implementation of non-animal methods in infection research, in line with 3R principles.