Cloning, expression, and preliminary characterization of the dysferlin tegument protein in Schistosoma japonicum

• Published in: Parasitology international 2013-12, Vol.62 (6), p.522-529
• Main Authors: Xiong, Yanian, Ai, Dezhou, Meng, Peipei, Wei, Meimei, Hong, Yang, Zhang, Min, Huang, Lini, Fu, Zhiqiang, Shi, Yaojun, Lin, Jiaojiao
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Helminth - immunology; Cell Membrane - metabolism; Cloning, Molecular; DNA, Complementary - chemistry; DNA, Complementary - genetics; DNA, Helminth - chemistry; DNA, Helminth - genetics; Dose-Response Relationship, Drug; Down-Regulation; Dysferlin; Escherichia coli; Female; Gastroenterology and Hepatology; Gene Expression Regulation; Helminth Proteins - genetics; Helminth Proteins - metabolism; Infectious Disease; Lung - parasitology; Male; Mice; Mice, Inbred BALB C; Plasma membrane repair; Recombinant Proteins; RNA, Messenger - genetics; Schistosoma japonicum; Schistosoma japonicum - genetics; Schistosoma japonicum - immunology; Schistosoma japonicum - metabolism; Sequence Analysis, DNA; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• ISSN: 1383-5769; 1873-0329
• DOI: 10.1016/j.parint.2013.07.008

Abstract The schistosomal tegument is a dynamic host-interactive layer. Proteins exposed to the host on the tegumental surface are important for completion of the parasitic lifecycle. Dysferlin is a member of the ferlin family and is involved in plasma membrane repair. Based on the results of a proteomics study of tegument surface proteins of Schistosoma japonicum in our laboratory, dysferlin was identified as a tegumental protein of S. japonicum . The gene encoding S. japonicum dysferlin (SjDF), which codes for several Ca 2 + binding sites, was cloned, expressed in Escherichia coli , and characterized. Western blot analysis revealed that recombinant SjDF had good immunogenicity. Real-time RT-PCR analysis showed that SjDF was upregulated mainly in adult worms and the transcription level in 42-day-old female worms was significantly higher than that in males. Immunofluorescence analysis revealed that SjDF was mainly distributed in the tegument at various developmental stages. Experimental mice were treated with praziquantel and at 35 days post-infection, we noted that damage to the tegument and subtegument worsened and did not recover at 36 h post-treatment in the high-dose group and was accompanied by downregulation of SjDF mRNA, while the damage was less severe and recovered by this time in the low-dose group, and accompanied by upregulation of SjDF. Our results suggested that SjDF is a tegumental protein that may be important in schistosomal development and may participate in the repair process in muscle and tegument, and could present a viable vaccine candidate for schistosomiasis.