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Specific in situ immuno-imaging of pulmonary-resident memory lymphocytes in human lungs
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Specific in situ immuno-imaging of pulmonary-resident memory lymphocytes in human lungs
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Specific in situ immuno-imaging of pulmonary-resident memory lymphocytes in human lungs
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Journal Article
Specific in situ immuno-imaging of pulmonary-resident memory lymphocytes in human lungs
2023
Overview
Pulmonary-resident memory T cells (T
) and B cells (B
) orchestrate protective immunity to reinfection with respiratory pathogens. Developing methods for the
detection of these populations would benefit both research and clinical settings.
To address this need, we developed a novel
immunolabelling approach combined with clinic-ready fibre-based optical endomicroscopy (OEM) to detect canonical markers of lymphocyte tissue residency
in human lungs undergoing
lung ventilation (EVLV).
Initially, cells from human lung digests (confirmed to contain T
/B
populations using flow cytometry) were stained with CD69 and CD103/CD20 fluorescent antibodies and imaged
using KronoScan, demonstrating it's ability to detect antibody labelled cells. We next instilled these pre-labelled cells into human lungs undergoing EVLV and confirmed they could still be visualised using both fluorescence intensity and lifetime imaging against background lung architecture. Finally, we instilled fluorescent CD69 and CD103/CD20 antibodies directly into the lung and were able to detect T
/B
following
labelling within seconds of direct
delivery of microdoses of fluorescently labelled antibodies.
, no wash, immunolabelling with
OEM imaging is a novel methodology with the potential to expand the experimental utility of EVLV and pre-clinical models.
