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Despite decades of ongoing research, single cell data within the microenvironment of granulomatous inflammatory reactions to Mycobacterium tuberculosis (Mtb) remain largely a mystery. Similarly, because of its recent emergence and rapid mutation, SARS-CoV-2 is still a major pandemic and remains a highly studied pathogen. To continue seeking novel information about these diseases, new research ... read moremodalities must be developed and validated. With advances in protein and RNA multiplexing and tissue-based cyclic immunofluorescence (t-CyCIF), single slides of fixed tissue can be stained with dozens of markers at a time to elucidate new information on regional transcriptomics, immunophenotypes, and cellular interactions. With success using these modalities in oncology research, we aimed to adapt this imaging method for animal models of Mtb and SARS-CoV-2 infections to better understand tissue responses in the context of natural infections, vaccination, and host-directed therapies for these globally important diseases. We designed and validated a 15-marker multiplexed immunofluorescence panel to immunophenotype immune cells in lung, spleen, and lymph node of rhesus macaques as a function of geospatial proximity to pathological regions of interest.
Cummings School Summer Scholar 2021.read less
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