OR10

1: Department of Histology, Research Center Borstel, Germany; 2: National Institute for Tuberculosis and Lung Diseases, Tbilisi, Georgia; 3: University of Georgia; 4: Office of Cyber Infrastructure and Computational Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, Bethesda, Maryland, USA; 5: Cellular Microbiology, Research Center Borstel, Germany; 6: Thematic Translational Unit Tuberculosis, German Center for Infection Research – Borstel Site

A detailed understanding of host-pathogen interactions is essential to develop innovative strategies against multi-drug-resistant tuberculosis (MDR-TB). To investigate pathogenic mechanisms at the site of infection, we assembled a cohort of partial lung resections from patients with drug-susceptible, MDR, and XDR tuberculosis. Lung tissue was stratified based on pathological patterns—such as unaffected areas, diffuse inflammation, cellular granulomas, and necrotic granulomas—and tissue microarrays were constructed to integrate multiple patient samples and lesion types within single paraffin blocks. Using multispectral imaging, we identified key immunological regions at the protein level, including multinucleated giant cell areas, T cell- and macrophage-rich zones, early and late-stage granulomas, and tertiary lymphoid structures. Spatial transcriptomics was then employed to obtain region-specific gene expression profiles, which were correlated with distinct immunological functions. For example, macrophage-rich regions displayed both type I and type II interferon responses, while giant cell areas showed gene signatures related to pH regulation and acidification. The outer rim of granulomas—enriched in T and B cells—were characterized by growth factor signaling and complement activation pathways. These findings reveal distinct molecular signatures within defined morphological niches that are associated with either protective or pathological outcomes. Our results shed light on the immunometabolic landscape of pulmonary TB lesions and may inform the development of targeted, host-directed therapies.