Intravital Imaging of Adoptive T-Cell Morphology, Mobility and Trafficking Following Immune Checkpoint Inhibition in a Mouse Melanoma Model.

Autor:	Lau D; Cancer Research UK Cambridge Centre, Cambridge, United Kingdom.; Department of Radiology, University of Cambridge, Cambridge, United Kingdom., Garçon F; Laboratory of Lymphocyte Signaling and Development, The Babraham Institute, Cambridge, United Kingdom., Chandra A; Department of Medicine, University of Cambridge, Cambridge, United Kingdom.; Department of Pathology, University of Cambridge, Cambridge, United Kingdom., Lechermann LM; Department of Radiology, University of Cambridge, Cambridge, United Kingdom., Aloj L; Cancer Research UK Cambridge Centre, Cambridge, United Kingdom.; Department of Radiology, University of Cambridge, Cambridge, United Kingdom.; Department of Nuclear Medicine, Addenbrooke's Hospital, Cambridge, United Kingdom., Chilvers ER; Department of Medicine, University of Cambridge, Cambridge, United Kingdom., Corrie PG; Department of Oncology, Addenbrooke's Hospital, Cambridge, United Kingdom., Okkenhaug K; Department of Pathology, University of Cambridge, Cambridge, United Kingdom., Gallagher FA; Cancer Research UK Cambridge Centre, Cambridge, United Kingdom.; Department of Radiology, University of Cambridge, Cambridge, United Kingdom.
Jazyk:	angličtina
Zdroj:	Frontiers in immunology [Front Immunol] 2020 Jul 22; Vol. 11, pp. 1514. Date of Electronic Publication: 2020 Jul 22 (Print Publication: 2020).
DOI:	10.3389/fimmu.2020.01514
Abstrakt:	Efficient T-cell targeting, infiltration and activation within tumors is crucial for successful adoptive T-cell therapy. Intravital microscopy is a powerful tool for the visualization of T-cell behavior within tumors, as well as spatial and temporal heterogeneity in response to immunotherapy. Here we describe an experimental approach for intravital imaging of adoptive T-cell morphology, mobility and trafficking in a skin-flap tumor model, following immune modulation with immune checkpoint inhibitors (ICIs) targeting PD-L1 and CTLA-4. A syngeneic model of ovalbumin and mCherry-expressing amelanotic mouse melanoma was used in conjunction with adoptively transferred OT-1 + cytotoxic T-cells expressing GFP to image antigen-specific live T-cell behavior within the tumor microenvironment. Dynamic image analysis of T-cell motility showed distinct CD8 + T-cell migration patterns and morpho-dynamics within different tumor compartments in response to ICIs: this approach was used to cluster T-cell behavior into four groups based on velocity and meandering index. The results showed that most T-cells within the tumor periphery demonstrated Lévy-like trajectories, consistent with tumor cell searching strategies. T-cells adjacent to tumor cells had reduced velocity and appeared to probe the local environment, consistent with cell-cell interactions. An increased number of T-cells were detected following treatment, traveling at lower mean velocities than controls, and demonstrating reduced displacement consistent with target engagement. Histogram-based analysis of immunofluorescent images from harvested tumors showed that in the ICI-treated mice there was a higher density of CD31 + vessels compared to untreated controls and a greater infiltration of T-cells towards the tumor core, consistent with increased cellular trafficking post-treatment. (Copyright © 2020 Lau, Garçon, Chandra, Lechermann, Aloj, Chilvers, Corrie, Okkenhaug and Gallagher.)
Databáze:	MEDLINE
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