Toxoplasma gondii is an obligate intracellular protozoan parasite that can infect all nucleated cells through active invasion. Some non-canonical pathways for T. gondii infection of macrophages have recently been reported. We report a new mode of T. gondii invasion using a time-lapse imaging system, in which T. gondii tachyzoites are engulfed by a tube-like structure on peritoneal macrophage phagosomes and then escape from the phagosomes. Escaped parasites re-invade macrophages through intercellular junctions between their apical end and host cell membranes. We call this invasion pathway of T. gondii “pseudopod-assisted invasion” (PAI). The completion of this invasion process depends on parasitic motility and secretion of adhesins from parasitic micronemes. Our results provide new information about T. gondii infection and establish another platform for studying interactions between T. gondii and macrophages.

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