RESUMO
BACKGROUND: Taenia multiceps is a cestode parasite with its larval stage (metacestode), Coenurus cerebralis, mainly encysts in the central nervous system of sheep and other livestock causing cerebralis coenurosis. Since, treatment of coenurosis with chemotherapy showed little effect and surgical removal of cysts is not advisable in field conditions, vaccination is useful to control coenurosis. Previous study indicated that immunization with T. multiceps metacestode antigens could induce protection in sheep against coenurosis, so the aim of this study was to identify T. multiceps metacestode antigens in order to find potential vaccine development candidates for further study. METHODS: The protein extracts from the larval T. multiceps were analyzed by two-dimensional electrophoresis (2-DE) and characterized by mass spectrometry. RESULTS: A total of 150 protein spots were detected with isoelectric point (pI) value from 4.97 to 9.65 and molecular weight from 14 to 98 kDa. Twenty-two protein identities were determined by mass spectrometry and 15 unique proteins were obtained. Functional annotation revealed that some of these proteins are involved in catalytic activity, binding, metabolic, cellular process and stress response. Among these molecules are antioxidant proteins (peroxiredoxin and glutathione-S-transferase), glycolytic enzymes (malate dehydrogenase and enolase), proteins with chaperone activity (heat shock protein 70 and small heat shock protein), and structural proteins (actin, actin modulator protein and paramyosin). CONCLUSION: The identification of T. multiceps metacestode protein will provide valuable information to elucidate their specific roles in the parasitism and screen new targets for vaccine development.
RESUMO
Protoscoleces of Taenia multiceps were collected from the naturally infected sheep and total RNA was extracted. Specific primers were designed according to TaHe2-D11 mRNA sequence and T. multiceps thioredoxin peroxidase gene (TmTPx) was amplified by RT-PCR. PCR products were ligated into pMD18-T vector and transformed to E. coli DH5alpha. The recombinant plasmids were identified by restriction digestion and sequencing. A 614 bp cDNA was amplified. The TmTPx open reading frame (591 bp) encoded a 196-amino acid protein with Mr 21,690, pI 7.61. Bioinformatics analysis indicated that TmTPx had a typical 2-Cys Prx conserved domain. Phylogenetic tree revealed that T. multiceps had the closest relationship to T. asiatica, followed by T. solium and T. crassiceps, E. granulosus and E. multilocularis.
