title: Cloning, expression, and characterization of Schistosoma japonicum tegument protein phosphodiesterase-5.
abstract: The tegument proteins of schistosomes are regarded as potential vaccine candidates and drug targets to control schistosomiasis. Nucleotide pyrophosphatase/phosphodiesterase-5 (NPP-5), which belongs to a multigene family of nucleotide pyrophosphatase/phosphodiesterases (NPPs), is important in the hydrolysis of pyrophosphate or phosphodiester bonds in nucleotides and their derivatives. In the present study, SjNPP-5, identified as one of the tegument proteins of Schistosoma japonicum in our previous proteomic studies, was cloned on a fragment of 1,371 bp and expressed as a recombinant protein of 69 kDa. Real-time RT-PCR analysis showed that SjNPP-5 was up-regulated at 21-42 days, and the expression level in 42-day-old male worms was almost nine times higher than that in females. Western blot analysis revealed that rSjNPP-5 had good antigenicity. Immunofluorescence analysis found that SjNPP-5 was a membrane-associated antigen mainly distributed on the surface of the male adult worm of S. japonicum. BALB/c mice vaccinated with rSjNPP-5 three times showed a 29.90% worm reduction (P < 0.05) and a 26.21% egg count reduction (P > 0.05). Immunization with rSjNPP-5 induced a mixed Th1/Th2 response in which Th1 was dominant. The response was characterized by a reduced IgG1/IgG2a ratio and elevated production of cytokines IFN-γ and IL-4. This study suggested that SjNPP-5 may be important in schistosome development, and further investigations are required to fully understand the function of this molecule.
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of Agriculture, 518 Ziyue road, Minhang, Shanghai 200241, People's Republic of China.
authors.forename: Min
authors.initials: M
authors.lastname: Zhang
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Han
authors.forename: Zhu
authors.initials: Z
authors.lastname: Zhu
authors.forename: Dong
authors.initials: D
authors.lastname: Li
authors.forename: Yang
authors.initials: Y
authors.lastname: Hong
authors.forename: Xiujuan
authors.initials: X
authors.lastname: Wu
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Fu
authors.forename: Jiaojiao
authors.initials: J
authors.lastname: Lin
mesh_terms: D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015153:Blotting, Western; D003001:Cloning, Molecular; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5 / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D011817:Rabbits; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012549:Schistosoma japonicum / Q000201:enzymology* / Q000235:genetics; D012554:Schistosomiasis japonica / Q000276:immunology / Q000517:prevention & control; D014611:Vaccination / Q000379:methods
chemical_list: D000947:Antigens, Helminth; D015801:Helminth Proteins; D011994:Recombinant Proteins; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5