Leishmania amazonensis amastigotes invade non-phagocytic cells via highly localized parasite-induced actin remodeling.

Intracellular parasites are pathogens that must invade and persist within host cells. In the case of Leishmania spp., it is generally assumed that the parasite must be phagocytosed to further establish residence within professional phagocytic cells. However, several studies report the presence of Leishmania spp. in non-phagocytic cells, highlighting their ability to enter cells independently of classical phagocytosis. Indeed, we have recently demonstrated that promastigotes, the infective form transmitted to hosts during the bite of the insect vector, subvert a lysosome-exocytosis-dependent membrane repair pathway to invade fibroblasts. Here, we investigate the invasion of non-phagocytic cells by L. amazonensis amastigotes, the infective form directly responsible for host-to-vector transmission, disease generation and parasite dissemination during infection in humans and other mammalian hosts. Our results show that amastigotes rapidly induce their entry into cells lacking classical phagocytic capability, where they survive, multiply, and persist. Invasion depends on parasite-induced actin remodeling confined to the parasite-host plasma membrane contact site, with localized recruitment of Rho GTPases that fuel actin polymerization at invasion foci. Our findings highlight the remarkable ability of Leishmania amastigotes to induce their own internalization into virtually any cell type, bypassing the need for conventional phagocytosis. This property may profoundly influence parasite biology by enabling amastigotes to cross cellular barriers, disseminate, and silently establish infection in diverse host cells. Importantly, when considering non-phagocytic cells, our results demonstrate that Leishmania amazonensis employs distinct, stage-specific invasion mechanisms: promastigotes co-opt host cell lysosomes, whereas amastigotes depend on F-actin dynamics.