Molecular characterization of a Trichinella spiralis serine proteinase
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RESEARCH ARTICLE
Open Access
Molecular characterization of a Trichinella spiralis serine proteinase Xin Yue, Xiang Yuan Sun, Fang Liu, Chen Xi Hu, Ying Bai, Qi Da Yang, Ruo Dan Liu, Xi Zhang, Jing Cui* and Zhong Quan Wang*
Abstract The aim of this study was to investigate the biological characteristics and functions of a Trichinella spiralis serine proteinase (TsSerp) during larval invasion and development in the host. The full-length TsSerp cDNA sequence was cloned and expressed in Escherichia coli BL21. The results of RT-PCR, IFA and western blotting analyses showed that TsSerp was a secretory protein that was highly expressed at the T. spiralis intestinal infective larva and muscle larva stages and primarily located at the cuticle, stichosome and intrauterine embryos of the parasite. rTsSerp promoted the larval invasion of intestinal epithelial cells (IECs) and the enteric mucosa, whereas an anti-rTsSerp antibody impeded larval invasion; the promotion and obstruction roles were dose-dependently related to rTsSerp and the antirTsSerp antibodies, respectively. Vaccination of mice with rTsSerp elicited a remarkable humoral immune response (high levels of serum IgG, IgG1/IgG2a, IgE and IgM), and it also triggered both systemic (spleen) and local intestinal mucosal mesenteric lymph node (MLN) cellular immune responses, as demonstrated by a significant elevation in Th1 cytokines (IFN-γ) and Th2 cytokines (IL-4) after the spleen and MLN cells from vaccinated mice were stimulated with rTsSerp. Anti-TsSerp antibodies participated in the killing and destruction of newborn larvae via ADCC. The mice vaccinated with rTsSerp exhibited a 48.7% reduction in intestinal adult worms and a 52.5% reduction in muscle larvae. These results indicated that TsSerp participates in T. spiralis invasion and development in the host and might be considered a potential candidate target antigen to develop oral polyvalent preventive vaccines against Trichinella infection. Keywords: Trichinella spiralis, serine proteinase, invasion, intestinal epithelial cells (IECs), immunogenicity, ADCC Introduction The genus Trichinella, including 10 species and 3 genotypes, is distributed in more than 150 domestic and wild mammals, and some species of the genus may cause the meat-borne zoonosis trichinellosis [1]. Humans acquire trichinellosis by consuming raw or semi-raw meat contaminated with Trichinella infective larvae. In China, endemic areas of human trichinellosis are mainly located in the southwestern region, but Trichinella infection in humans and animals was found in 33 of 34 provinces [2]. From 2004 to 2009, 12 trichinellosis outbreaks due *Correspondence: [email protected]; [email protected] Department of Parasitology, Medical College, Zhengzhou University, Zhengzhou 450052, China
to infected pork were reported [3]. Domestic pork is still the primary infectious source of Trichinella infection in developing countries [4–6]. Because Trichinella is a zoonotic parasite and has a broad distribution of natural hosts in the world, it is difficult to
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