The complement system is a pivotal component of innate immunity, extensively studied in vertebrates but also present in invertebrates. This study explores the existence of a terminal complement pathway in the tunicate Botryllus schlosseri, aiming to understand the evolutionary integration of innate and adaptive immunity. Through transcriptome analysis, we identified a novel transcript, BsITCCP, encoding a protein with both MACPF and LDLa domains—a structure resembling that of vertebrate C9 but with a simpler organization. Phylogenetic reconstruction positions BsITCCP between invertebrate perforins and vertebrate terminal complement proteins, suggesting an evolutionary link. Localization studies confirmed that bsitccp is transcribed in cytotoxic morula cells (MCs), which are also responsible for producing other complement components like BsC3, BsMBL, BsMASP, and BsBf. Functional assays demonstrated that bsitccp transcription is upregulated in response to nonself challenges and is dependent on BsC3 activity; inhibition of BsC3 led to a significant reduction in BsITCCP expression. Electron microscopy revealed that MCs form contact with perforated yeast cells, indicating a possible mechanism of cell lysis similar to the immunological synapse observed in vertebrates. These findings suggest that a C3-governed lytic complement pathway exists in B. schlosseri, challenging the assumption that a C5 ortholog is necessary for such a pathway. This work enhances our understanding of the evolution of the complement system and suggests that invertebrates possess a terminal complement complex capable of mediating cell lysis, regulated by C3. Future studies will focus on confirming the pore-forming ability of BsITCCP and its role in the immunological synapse.

C3 is the central effector of the complement system and one of the most conserved molecules of metazoan innate immunity. Despite its relevance, little is known about complement components in Lophotrochozoa. Here, we characterise the C3 ortholog of the freshwater gastropod Pomacea canaliculata (PcC3) using long-read transcriptomics, phylogenetics, expression profiling and immunohistochemistry. PcC3 displays a fully conserved domain architecture and clusters phylogenetically in agreement with metazoan relationships. Following LPS challenge, PcC3 is up-regulated in haemocytes and unexpectedly in the ampulla, but not in the posterior kidney, revealing functional specialization of immune tissues. Early developmental transcriptomes reveal strong and sustained PcC3 activation from 3 to 9 dpf. These data suggest that PcC3 retains canonical immune functions but may also contribute to developmental and homeostatic cell clearance, supporting P. canaliculata as a valuable model for complement biology in molluscs.