Molecular tracking devices quantify antigen distribution and archiving in the murine lymph node.

Abstract

The detection of foreign antigens in vivo has relied on fluorescent conjugation or indirect read-outs such as antigen presentation. In our studies, we found that these widely used techniques had several technical limitations that have precluded a complete picture of antigen trafficking or retention across lymph node cell types. To address these limitations, we developed a 'molecular tracking device' to follow the distribution, acquisition, and retention of antigen in the lymph node. Utilizing an antigen conjugated to a nuclease-resistant DNA tag, acting as a combined antigen-adjuvant conjugate, and single-cell mRNA sequencing, we quantified antigen abundance in the lymph node. Variable antigen levels enabled the identification of caveolar endocytosis as a mechanism of antigen acquisition or retention in lymphatic endothelial cells. Thus, these molecular tracking devices enable new approaches to study dynamic tissue dissemination of antigen-adjuvant conjugates and identify new mechanisms of antigen acquisition and retention at cellular resolution in vivo.

• Authors: Shannon M Walsh, Ryan M Sheridan, Erin D Lucas, Thu A Doan, Brian C Ware, Johnathon Schafer, Rui Fu, Matthew A Burchill, Jay R Hesselberth, Beth Ann Jiron Tamburini
• Journal: eLife (Elife) Vol. 10 (Apr 12 2021) ISSN: 2050-084X [Electronic] England
• PMID: 33843587 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: © 2021, Walsh et al.

Chemical References

• Antigens
• OVA-8
• Peptide Fragments
• Phosphorothioate Oligonucleotides
• RNA, Messenger
• Ovalbumin
• DNA

Topics

• Animals
• Antigen Presentation
• Antigens (genetics, immunology, metabolism)
• Caveolae (immunology, metabolism)
• Cells, Cultured
• DNA (genetics, metabolism)
• Dendritic Cells (immunology, metabolism)
• Endocytosis
• Endothelial Cells (immunology, metabolism)
• Lymph Nodes (immunology, metabolism)
• Macrophages (immunology, metabolism)
• Mice, Inbred C57BL
• Mice, Transgenic
• Ovalbumin (genetics, immunology, metabolism)
• Peptide Fragments (genetics, immunology, metabolism)
• Phosphorothioate Oligonucleotides (genetics, metabolism)
• RNA, Messenger (genetics, metabolism)
• Sequence Analysis, RNA
• Single-Cell Analysis
• Time Factors
• Tissue Distribution
• Transcriptome
• Mice