Kiselev, A.; Schmitter-Sanchez, A. D.; Williams, S.; Park, S.

Langerhans cells (LCs) are a specialized subset of dendritic cells residing in the epidermis, where they form a dense network that functions as a frontline defense by detecting environmental antigens and sensing skin damage. Despite extensive research, many immunological and physiological aspects of LCs function remain poorly understood. In this study, we combined single-cell transcriptomic analysis and intravital imaging to reconstruct the activation trajectory of epidermal LCs and identified a distinct intermediate state that precedes their migration. We present a high-resolution single-cell transcriptomic dataset of over 22,000 high-quality epidermal LCs under both injured and homeostatic conditions. We found similar LCs responses to chemical and physical skin injuries. Our analysis defines specific LCs subpopulations representing sequential activation stages, characterized at the level of pathways and transcription factors. Notably, we identified a previously unrecognized population of activated LCs distinguished by the expression of complement system components and receptors. Integrating our data with an external dataset of wounded and unwounded skin revealed that wound-specific, WNT-modulated fibroblasts are the primary source of C3, the central component of the complement cascade. Intravital imaging of C3-deficient mice demonstrated that C3 is essential for the effective recruitment of activated LCs to wound sites. Together, these findings uncover a novel population of activated epidermal LCs and highlight complement signaling as a critical mediator of LCs recruitment during skin injury.