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Gut microbiota is known to educate the immune system, balancing effector and regulatory T cells responses, but it remains unknown how bacteria-specific T cells develop. In this webinar, Dr. Daniel Zegarra-Ruiz of Memorial Sloan Kettering Cancer Center, recipient of the AAI 2020 Lefrançois-BioLegend Memorial Award, discusses how thymic T cell development is altered after colonization with model intestinal microbes. These findings include thymic expansion of bacteria-specific naive T cells, discovery of bacterial DNA in the thymus, enrichment of CX3CR1+ dendritic cells (CX3+ DCs), and the effect of depleting the gut microbiota with antibiotics. Thymic microbiota-specific T cells were found to differentiate into intestinal effector T cells, offering protection or mediating pathology. Together, Dr. Zegarra-Ruiz’s data suggests that trafficking of bacteria to the thymus by intestinal DCs drives T cell expansion of microbiota-specific T cells, shaping peripheral immunity against pathogens and the microbiota itself. What you will learn: • How to identify and characterize microbiota-specific T cells in the thymus • How gut microbial depletion affects thymic populations • How to study antigen-presenting cells traffic from the gut to the thymus • How to assess thymic microbiota-specific T cells expansion and differentiation