Schistosoma japonicum: cloning, expression and characterization of a gene encoding the α5-subunit of the proteasome.
The development of an effective vaccine against the schistosome is thought to be the most desirable means to control schistosomiasis, even though there is an effective means of chemotherapy with praziquantel. A full-length cDNA encoding the Schistosoma japonicum proteasome subunit alpha type 5 protein (SjPSMA5) was first isolated from 18-day-schistosomulum cDNAs. The cDNA had an open reading frame (ORF) of 747bp and encoded 248 amino acids. Real-time quantitative RT-PCR analysis revealed that SjPSMA5 is up-regulated in 18-day and 32-day schistosomes, and the level of expression in male is around fourfold higher than that in female worms at 42 days. The SjPSMA5 was subcloned into pET28a(+) and expressed as inclusion bodies in Escherichia coli BL21 (DE3) cells. Western blotting showed that the recombinant SjPSMA5 (rSjPSMA5) was immunogenic. After immunization of BALB/c mice with rSjPSMA5, reductions of 23.29% and 35.24% were obtained in the numbers of worms and eggs in the liver, respectively. The levels of specific IgG antibodies and CD(4)(+) cells were significantly higher (P<0.01) in the group vaccinated with rSjPSMA5 combined with Seppic 206 adjuvant than in the other groups, as detected by enzyme linked immunosorbent assay (ELISA) and flow cytometry. The study suggested that rSjPSMA5 induced partial immunoprotection against S. japonicum in BALB/c mice, and it could be a potential vaccine candidate against schistosomiasis.
Hong
Yang
Y
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, PR China.
Han
Hongxiao
H
Peng
Jinbiao
J
Li
Ye
Y
Shi
Yaojun
Y
Fu
Zhiqiang
Z
Liu
Jinming
J
Lin
Jiaojiao
J
Li
Xiangrui
X
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D015153:Blotting, Western; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D007111:Immunity, Cellular; D007074:Immunoglobulin G / Q000097:blood; D008099:Liver / Q000469:parasitology; D008297:Male; D051379:Mice; D010802:Phylogeny; D046988:Proteasome Endopeptidase Complex / Q000235:genetics* / Q000276:immunology; D018510:RNA, Helminth / Q000302:isolation & purification; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012549:Schistosoma japonicum / Q000145:classification / Q000235:genetics* / Q000276:immunology; D012554:Schistosomiasis japonica / Q000276:immunology / Q000517:prevention & control; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000276:immunology
D000909:Antibodies, Helminth; D007074:Immunoglobulin G; D018510:RNA, Helminth; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D046988:Proteasome Endopeptidase Complex