Identification and profiling of circulating antigens by screening with the sera from schistosomiasis japonica patients.
BACKGROUND
Schistosomiasis is a chronic disease caused by trematode flatworms of the genus Schistosoma. The disease remains a serious public health problem in endemic countries and affects at least 207 million people worldwide. A definite diagnosis of the disease plays a key role in the control of schistosomiasis. The detection of schistosome circulating antigens (CAs) is an effective approach to discriminate between previous exposure and current infection. Different methods have been investigated for detecting the CAs. However, the components of the schistosome CAs remain unclear. In this study, we analyzed the CAs in sera of patients infected with Schistosoma japonicum.


METHODS
The parasites were collected from the infected rabbits for preparing the adult worm antigen (AWA). The hyline hens were immunized subcutaneously with AWA to produce anti-AWA IgY. The IgY was purified by water-dilution and ammonium sulfate precipitation method and identified by ELISA and Western blotting. After purification and characterization, IgY was immobilized onto the resin as a capture antibody. The circulating antigens were immune-precipitated from patients' serum samples by direct immunoprecipitation. The precipitated proteins were separated by one-dimensional electrophoresis and analyzed by LC-MS/MS.


RESULTS
Firstly, the IgY against AWA was produced from the eggs of immunized hens by AWA, which gave a titer of 1:12800. The purified IgY was used as the capture antibody to enrich the CAs in sera of S. japonicum infected patients through immunoprecipitation. The CAs were determined by LC-MS/MS. There were four proteins, including protein BUD31 homolog, ribonuclease, SJCHGC06971 protein and SJCHGC04754 protein, which were identified among the CAs.


CONCLUSIONS
We developed a novel method based on IgY for identification and profiling CAs in sera of S. japonicum infected patients. Four new CAs were identified and have potential value for further development of an antigen assay.
School of Biotechnology, East China University of Science and Technology, Shanghai 200237, P,R, China.
Yan
Y
Lu
Bin
B
Xu
Chuan
C
Ju
Xiaojin
X
Mo
Shenbo
S
Chen
Zheng
Z
Feng
Xiaoning
X
Wang
Wei
W
Hu
D000818:Animals; D000947:Antigens, Helminth / Q000097:blood* / Q000276:immunology; D002645:Chickens; D006801:Humans; D007136:Immunoglobulins / Q000276:immunology; D047468:Immunoprecipitation / Q000379:methods; D013058:Mass Spectrometry; D011817:Rabbits; D012549:Schistosoma japonicum / Q000378:metabolism*; D012554:Schistosomiasis japonica / Q000097:blood / Q000469:parasitology*
D000947:Antigens, Helminth; C016512:IgY; D007136:Immunoglobulins