Abstract
Myoferlin is a member of the ferlin family of proteins, which are involved in plasma membrane repair, and has been identified as one of the tegument proteins of Schistosoma japonicum. The tegument proteins are potential candidates for vaccines and new drug targets. In this study, myoferlin of S. japonicum (SjMF) was cloned, expressed and characterized, the potential of SjMF recombinant protein (rSjMF) as a vaccine candidate was evaluated, and the effect of praziquantel on SjMF was detected by Real-time PCR. Immunofluorescence showed that this protein was mainly distributed on the surface of worms at different stages. Sequence analysis revealed that the SjMF open reading frame was conserved at all stages of the S. japonicum life cycle. And SjMF transcription was upregulated in 42-day-old worms, and was significantly higher in female worms. Western blotting revealed that rSjMF showed strong immunogenicity. The cytokine profile and IgG isotype analysis demonstrated that rSjMF plus ISA206 immunization induced a mixed T helper (Th)1/Th2 response. Purified rSjMF emulsified with ISA206 adjuvant significantly reduced worm burden from 21.8% to 23.21% and liver egg number from42.58% to 28.35%. Besides, SjMF transcription was downregulated when worms were exposed to low-dose praziquantel (PZQ) and upregulated when PZQ was degraded, accompanied by recovery of damaged tegument. When worms were exposed to high-dose PZQ, SjMF transcription was downregulated all the time and the damaged tegument did not recover. These findings indicated that SjMF is a potential vaccine against S. japonicum and provides the basis for further investigations into the biological function of SjMF.
Highlights
Schistosomes are parasitic blood helminths that infect millions of people in tropical and subtropical countries [1]
We described the cloning, expression, and immunolocalization of the myoferlin of S. japonicum (SjMF) gene, as well as the immunogenicity of recombinant SjMF
Molecular Cloning and Sequence Analysis of SjMF The sequence of the S. japonicum cDNA encoding SjMF was obtained by PCR amplification with specific oligonucleotides
Summary
Schistosomes are parasitic blood helminths that infect millions of people in tropical and subtropical countries [1]. 779 million people are at risk of being infected in 76 endemic countries and an estimated 280,000 deaths are directly or indirectly attributable to the disease annually [2,3]. Schistosomiasis control remains a major challenge in China. Schistosomiasis control strategy is mainly based on treatment of infected individuals with praziquantel (PZQ). PZQ can effectively reduce the morbidity associated with schistosomiasis, but it has been proved not to be sufficient to control disease transmission and prevent reinfection [4,5]. An effective vaccine against schistosomiasis would be essential to the current control strategy, mainly because it would provide long-lasting immunity against infection. It is important to identify the appropriate schistosomal antigens that could induce activity against schistosomal infection or reduce the release of live eggs to limit parasite transmission
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