Supramolecular Organization Predicts Protein Nanoparticle Delivery to Neutrophils for Acute Lung Inflammation Diagnosis and Treatment

Jacob W. Myerson; Priyal N. Patel; Nahal Habibi; Landis R. Walsh; Yi-Wei Lee; David C. Luther; Laura T. Ferguson; Michael H. Zaleski; Marco E. Zamora; Oscar A. Marcos-Contreras; Patrick M. Glassman; Ian Johnston; Elizabeth D. Hood; Tea Shuvaeva; Jason V. Gregory; Raisa Y. Kiseleva; Jia Nong; Kathryn M. Rubey; Colin F. Greineder; Samir Mitragotri; George S. Worthen; Vincent M. Rotello; Joerg Lahann; Vladimir R. Muzykantov; Jacob S. Brenner

For imaging neutrophil content in naïve and IV-LPS-injured lungs, mice were intravenously injected with rat anti-mouse anti-Ly6G antibody (clone 1A8) and sacrificed 30 minutes later. Lungs were embedded in M1 medium, flash frozen, and sectioned in 10 µm slices. Sections were stained with PerCP-conjugated anti-rat secondary antibody and neutrophil-associated fluorescence was observed with epifluorescence microscopy. Similar procedures enabled histological imaging of lysozyme-dextran nanogels in IV-LPS-injured lungs. Nanogels synthesized with rhodamine-dextran were administered intravenously in injured mice 30 minutes prior to sacrifice. Lungs were sectioned as above and stained with clone 1A8 anti-Ly6G antibody, followed by Briliant Violetconjugated anti-rat secondary antibody.