Folia Parasitologica 65:007 (2018) | DOI: 10.14411/fp.2018.007

Cloning, expression and characterisation of a cysteine protease from Trichinella spiralis

Yan Yan Song^*, Li Ang Wang^*, Hua Na Ren, Xin Qi, Ge Ge Sun, Ruo Dan Liu, Peng Jiang, Xi Zhang, Jing Cui, Zhong Quan Wang

Department of Parasitology, Medical College, Zhengzhou University, PR China

^* These authors contributed on the work equally.

Cysteine protease is a superfamily of widespread proteolytic enzymes and plays a major role in larval invasion, migration, exsheathing, survival and immune evasion in parasites. In the present study, the gene coding cysteine proteinase of the nematode Trichinella spiralis (Owen, 1835) was cloned into pQE-80L and subsequently expressed in E. coli JM109. The rTsCP was purified and its antigenicity was identified by Western blot and ELISA. Using anti-rTsCP serum the native TsCP was identified in muscle larval crude proteins. The results of quantitative real-time PCR and immunofluorescence test demonstrated that the TsCP was expressed in all stages of T. spiralis and located mainly in cuticle, stichosome and reproductive organs. The immunisation of mice with rTsCP elicited Th2-predominant immune responses. Anti-rTsCP antibodies could partially inhibit the in vitro larval invasion of intestinal epithelial cells and kill the newborn larvae by an antibody-dependent cell-mediated dose-dependent cytotoxicity. The vaccinated mice exhibited a 54% reduction of adults and a 33% reduction of muscle larvae following challenge infection. The results suggested that the TsCP might be an indispensable protein in Trichinella invasion, development and survival of T. spiralis in hosts, and could be a potential vaccine target against infection.

Keywords: Trichinellosis, cysteine protease, invasion, development

Received: November 14, 2017; Accepted: March 19, 2018; Published online: May 29, 2018  Show citation