TY - JOUR TI - Molecular tracking devices quantify antigen distribution and archiving in the murine lymph node AU - Walsh, Shannon M AU - Sheridan, Ryan M AU - Lucas, Erin D AU - Doan, Thu A AU - Ware, Brian C AU - Schafer, Johnathon AU - Fu, Rui AU - Burchill, Matthew A AU - Hesselberth, Jay R AU - Tamburini, Beth Ann Jiron A2 - Dustin, Michael L A2 - Rothlin, Carla V A2 - Dustin, Michael L VL - 10 PY - 2021 DA - 2021/04/12 SP - e62781 C1 - eLife 2021;10:e62781 DO - 10.7554/eLife.62781 UR - https://doi.org/10.7554/eLife.62781 AB - 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. KW - single-cell mRNA sequencing KW - lymph node KW - antigen processing KW - antigen archiving KW - lymphatic endothelial cell KW - dendritic cell JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -