Cloning, expression and characterisation of a cysteine protease from Trichinella spiralis
| Autor: | Ge Ge Sun, Xi Zhang, Jing Cui, Peng Jiang, Ruo Dan Liu, Zhong Quan Wang, Xin Qi, Yan Yan Song, Li Ang Wang, Hua Na Ren |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Male Antigenicity Trichinella spiralis Trichinella Gene Expression Microbiology 03 medical and health sciences Mice Immune system Western blot Cysteine Proteases medicine Escherichia coli Animals Cloning Molecular Mice Inbred BALB C biology medicine.diagnostic_test Proteolytic enzymes Helminth Proteins Sequence Analysis DNA biology.organism_classification Cysteine protease Specific Pathogen-Free Organisms 030104 developmental biology Larva biology.protein Parasitology Female Antibody Microorganisms Genetically-Modified |
| Zdroj: | Folia parasitologica. 65 |
| ISSN: | 0015-5683 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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