Quantifying Leukocyte Egress via Lymphatic Vessels from Murine Skin and Tumors

Maria M. Steele, Madeline J. Churchill, Alec P. Breazeale, Ryan S. Lane, Nicholas A. Nelson, Amanda Lund

Research output: Contribution to journalArticle

Abstract

Leukocyte egress from peripheral tissues to draining lymph nodes is not only critical for immune surveillance and initiation but also contributes to the resolution of peripheral tissue responses. While a variety of methods are used to quantify leukocyte egress from non-lymphoid, peripheral tissues, the cellular and molecular mechanisms that govern context-dependent egress remain poorly understood. Here, we describe the use of in situ photoconversion for quantitative analysis of leukocyte egress from murine skin and tumors. Photoconversion allows for the direct labeling of leukocytes resident within cutaneous tissue. Though skin exposure to violet light induces local inflammatory responses characterized by leukocyte infiltrates and vascular leakiness, in a head-to-head comparison with transdermal application of fluorescent tracers, photoconversion specifically labeled migratory dendritic cell populations and simultaneously enabled the quantification of myeloid and lymphoid egress from cutaneous microenvironments and tumors. The mechanisms of leukocyte egress remain a missing component in our understanding of intratumoral leukocyte complexity, and thus the application of the tools described herein will provide unique insight into the dynamics of tumor immune microenvironments both at steady state and in response to therapy.

Original languageEnglish (US)
JournalJournal of visualized experiments : JoVE
Issue number143
DOIs
StatePublished - Jan 7 2019

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Lymphatic Vessels
Tumors
Skin
Leukocytes
Tissue
Neoplasms
Tumor Microenvironment
Labeling
Dendritic Cells
Blood Vessels
Chemical analysis
Lymph Nodes
Light
Population

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Quantifying Leukocyte Egress via Lymphatic Vessels from Murine Skin and Tumors. / Steele, Maria M.; Churchill, Madeline J.; Breazeale, Alec P.; Lane, Ryan S.; Nelson, Nicholas A.; Lund, Amanda.

In: Journal of visualized experiments : JoVE, No. 143, 07.01.2019.

Research output: Contribution to journalArticle

Steele, Maria M. ; Churchill, Madeline J. ; Breazeale, Alec P. ; Lane, Ryan S. ; Nelson, Nicholas A. ; Lund, Amanda. / Quantifying Leukocyte Egress via Lymphatic Vessels from Murine Skin and Tumors. In: Journal of visualized experiments : JoVE. 2019 ; No. 143.
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