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Related Experiment Video

Updated: Nov 9, 2025

Quantifying Leukocyte Egress via Lymphatic Vessels from Murine Skin and Tumors
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Molecular tracking devices quantify antigen distribution and archiving in the murine lymph node.

Shannon M Walsh1, Ryan M Sheridan2, Erin D Lucas3,4

  • 1Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, United States.

Elife
|April 12, 2021
PubMed
Summary

Researchers developed a molecular tracking device to study antigen distribution in lymph nodes. This new method reveals mechanisms of antigen acquisition and retention at cellular resolution.

Keywords:
antigen archivingantigen processingcell biologydendritic cellimmunologyinflammationlymph nodelymphatic endothelial cellmousesingle-cell mRNA sequencing

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Area of Science:

  • Immunology
  • Molecular Biology
  • Cell Biology

Background:

  • Current methods for detecting foreign antigens in vivo, such as fluorescent conjugation and antigen presentation assays, have limitations in fully characterizing antigen trafficking and retention within lymph node cell types.
  • A comprehensive understanding of antigen dynamics is crucial for developing effective vaccines and immunotherapies.

Purpose of the Study:

  • To develop a novel molecular tracking device for precise in vivo tracking of antigen distribution, acquisition, and retention within lymph nodes.
  • To overcome the limitations of existing techniques and provide a detailed cellular-level view of antigen dynamics.

Main Methods:

  • Developed a 'molecular tracking device' by conjugating an antigen to a nuclease-resistant DNA tag, creating a combined antigen-adjuvant conjugate.
  • Employed single-cell mRNA sequencing to quantify antigen abundance within individual lymph node cells.
  • Analyzed variable antigen levels to identify cellular mechanisms of antigen uptake and retention.

Main Results:

  • The molecular tracking device successfully monitored the distribution, acquisition, and retention of antigens in the lymph node.
  • Identified caveolar endocytosis as a key mechanism for antigen acquisition and retention in lymphatic endothelial cells.
  • Demonstrated the ability to quantify antigen abundance at a single-cell level, providing unprecedented resolution.

Conclusions:

  • The developed molecular tracking devices offer a powerful new approach for studying the dynamic dissemination of antigen-adjuvant conjugates in tissues.
  • This technology enables the discovery of novel mechanisms of antigen acquisition and retention at the cellular level in vivo.
  • Provides a foundation for future research into immune responses and the development of targeted immunotherapies.