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, and Beth A. Jirón Tamburini
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 thathave precluded a complete picture of antigen trafficking or retention across lymph nodecell types. To address these limitations, we developed a “molecular tracking device” to follow the distribution, acquisition, and retention of antigen in the lymph node. Utilizingan 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 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.
Data have been deposited at NCBI GEO
A cell browser is availble here