MUB₄₀ Binds to Lactoferrin and Stands as a Specific Neutrophil Marker

Neutrophils represent the most abundant immune cells recruited to inflamed tissues. A lack of dedicated tools has hampered their detection and study. We show that a synthesized peptide, MUB₄₀, binds to lactoferrin, the most abundant protein stored in neutrophil-specific and tertiary granules. Lactoferrin is specifically produced by neutrophils among other leukocytes, making MUB₄₀ a specific neutrophil marker. Naive mammalian neutrophils (human, guinea pig, mouse, rabbit) were labeled by fluorescent MUB₄₀ conjugates (-Cy5, Dylight405). A peptidase-resistant retro-inverso MUB₄₀ (RI-MUB₄₀) was synthesized and its lactoferrin-binding property validated. Neutrophil lactoferrin secretion during in vitro Shigella infection was assessed with RI-MUB₄₀-Cy5 using live cell microscopy. Systemically administered RI-MUB₄₀-Cy5 accumulated at sites of inflammation in a mouse arthritis inflammation model in vivo and showed usefulness as a potential tool for inflammation detection using non-invasive imaging. Improving neutrophil detection with the universal and specific MUB₄₀ marker will aid the study of broad ranges of inflammatory diseases. MUB₄₀ is a universal and specific marker of mammalian neutrophils. MUB₄₀ interacts with lactoferrin, stored in specific (β1), tertiary (β2), and secretome (γ) granules secreted upon stimulation. Neutrophil detection in inflamed or infected tissues is facilitated by MUB₄₀ peptides conjugated to fluorophores. Neutrophilic inflammation detection may be envisaged by combining MUB₄₀ with non-invasive imaging.