An Excretory Protein of Echinococcus multilocularis Inhibits Complement Classical Pathway Activation

Yiwen Qiu, Shu Shen, Yi Yang, Wentao Wang Department of Liver Surgery and Liver Transplantation Center, West China Hospital of Sichuan University, Chengdu, People’s Republic of ChinaCorrespondence: Wentao Wang, Department of Liver Surgery and Liver Transplantation Center, West China Hospital of Sichuan University, 37 Guoxue Road, Chengdu, 610041, People’s Republic of China, Tel +86 18980601895, Fax +86-028-85422871, Email wwtdoctor02@163.comIntroduction: Alveolar echinococcosis is a lethal zoonosis caused by Echinococcus multilocularis (E.m) larvae. The mechanism by which E.m evades host immune attacks and ensures long-term survival remains unexplained. The complement system is a cascade of sequentially activated complement proteins that results in opsonization-related phagocytosis or membrane lysis of invading organisms. Excretory/secretory proteins (ESPs) of parasites are the main antigens that induce the immune response and play important roles in the long-term survival.Methods: We investigated the possibility that E.m inhibits complement activation through ESPs and examined the potential related mechanism. A haemolysis assay was used to determine if and how in vitro culture medium of E.m containing ESPs can inhibit complement activation. Potential ESPs were annotated using bioinformatics methods, and one ESP was subsequently expressed as a recombinant protein with a eukaryotic expression system. The ability of this protein to inhibit complement activation was also tested by haemolysis assay.Results: These assays showed that in vitro culture medium of E.m inhibited activation of the complement classical pathway. EmuJ_000439500 encodes a protein containing seven Sushi domains, which was the only potential E.m-derived complement inhibitor (Em-CI, UniProt: A0A068Y4F2) annotated among the 653 ESPs. Recombinant Em-CI also displayed the ability to inhibit activation of the complement classical pathway.Discussion: The discovery of Em-CI sheds light on the mechanism by which E.m escapes killing by the complement system and provides potential targets for immunotherapy for parasitic diseases.Keywords: Echinococcus multilocularis, alveolar echinococcosis, complement inhibition, excretory/secretory protein