Identification and profiling of circulating antigens by screening with the sera from schistosomiasis japonica patients

Particle and Fibre Toxicology

Wei Hu¹  Xiaoning Wang¹  Zheng Feng³  Shenbo Chen³  Xiaojin Mo³  Chuan Ju³  Bin Xu³  Yan Lu² 

[1] Department of Microbiology and Microbial Engineering, School of LifeScience, Fudan University, Shanghai 200433, P.R. China;School of Biotechnology, East China University of Science and Technology, Shanghai 200237, P.R. China;National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Key Laboratory of Parasite and Vector biology, MOH; WHO Collaborating Center of Malaria, Schistosomiasis and Filariasis, Shanghai 200025, P.R. China

关键词: IgY;    Identification;    Circulating antigens;    Schistosoma japonicum;   

Others  :  1231782
DOI  :  10.1186/1756-3305-5-115

 received in 2011-11-30, accepted in 2012-05-17,  发布年份 2012

【 摘 要 】

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.

【 授权许可】

   
2012 Lu et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

【 参考文献 】

• [1]King CH: Toward the elimination of schistosomiasis. N Engl J Med 2009, 360:106-109.
• [2]Hotez PJ, Molyneux DH, Fenwick A, Kumaresan J, Sachs SE, Sachs JD, Savioli L: Control of neglected tropical diseases. N Engl J Med 2007, 357:1018-1027.
• [3]Hotez PJ, Bethony JM, Diemert DJ, Pearson M, Loukas A: Developing vaccines to combat hookworm infection and intestinal schistosomiasis. Nat Rev Microbiol 2010, 8:814-826.
• [4]Steinmann P, Keiser J, Bos R, Tanner M, Utzinger J: Schistosomiasis and water resources development: systematic review, meta-analysis, and estimates of people at risk. Lancet Infect Dis 2006, 6:411-425.
• [5]King CH, Dickman K, Tisch DJ: Reassessment of the cost of chronic helmintic infection: a meta-analysis of disability-related outcomes in endemic schistosomiasis. Lancet 2005, 365:1561-1569.
• [6]Balog CI, Alexandrov T, Derks RJ, Hensbergen PJ, van Dam GJ, Tukahebwa EM, Kabatereine NB, Thiele H, Vennervald BJ, Mayboroda OA, Deelder AM: The feasibility of MS and advanced data processing for monitoring Schistosoma mansoni infection. Proteomics Clin Appl 2010, 4:499-510.
• [7]van Lieshout L, Polderman AM, Deelder AM: Immunodiagnosis of schistosomiasis by determination of the circulating antigens CAA and CCA, in particular in individuals with recent or light infections. Acta Trop 2000, 77:69-80.
• [8]Zhou YB, Zheng HM, Jiang QW: A diagnostic challenge for Schistosomiasis japonica in China: consequences on praziquantel-based morbidity control. Parasit Vectors 2011, 4:194. BioMed Central Full Text
• [9]Midzi N, Butterworth AE, Mduluza T, Munyati S, Deelder AM, van Dam GJ: Use of circulating cathodic antigen strips for the diagnosis of urinary schistosomiasis. Trans R Soc Trop Med Hyg 2009, 103:45-51.
• [10]van Dam GJ, Wichers JH, Ferreira TM, Ghati D, van Amerongen A, Deelder AM: Diagnosis of schistosomiasis by reagent strip test for detection of circulating cathodic antigen. J Clin Microbiol 2004, 42:5458-5461.
• [11]Legesse M, Erko B: Field-based evaluation of a reagent strip test for diagnosis of schistosomiasis mansoni by detecting circulating cathodic antigen (CCA) in urine in low endemic area in Ethiopia. Parasite 2008, 15:151-155.
• [12]Kahama AI, Kremsner PG, van Dam GJ, Deelder AM: The dynamics of a soluble egg antigen of Schistosoma haematobium in relation to egg counts, circulating anodic and cathodic antigens and pathology markers before and after chemotherapy. Trans R Soc Trop Med Hyg 1998, 92:629-633.
• [13]Elliott DE: Schistosomiasis. Pathophysiology, diagnosis, and treatment. Gastroenterol Clin North Am 1996, 25:599-625.
• [14]Legesse M, Erko B: Field-based evaluation of a reagent strip test for diagnosis of Schistosoma mansoni by detecting circulating cathodic antigen in urine before and after chemotherapy. Trans R Soc Trop Med Hyg 2007, 101:668-673.
• [15]de Carvalho Souza A, Kuil J, Maljaars CE, Halkes KM, Vliegenthart JF, Kamerling JP: Synthesis and conjugation of oligosaccharide analogues of fragments of the immunoreactive glycan part of the circulating anodic antigen of the parasite Schistosoma mansoni. Org Biomol Chem 2004, 2:2972-2987.
• [16]Corstjens PL, van Lieshout L, Zuiderwijk M, Kornelis D, Tanke HJ, Deelder AM, van Dam GJ: Up-converting phosphor technology-based lateral flow assay for detection of Schistosoma circulating anodic antigen in serum. J Clin Microbiol 2008, 46:171-176.
• [17]Deelder AM, Qian ZL, Kremsner PG, Acosta L, Rabello AL, Enyong P, Simarro PP, van Etten EC, Krijger FW, Rotmans JP, et al.: Quantitative diagnosis of Schistosoma infections by measurement of circulating antigens in serum and urine. Trop Geogr Med 1994, 46:233-238.
• [18]Shane HL, Verani JR, Abudho B, Montgomery SP, Blackstock AJ, Mwinzi PN, Butler SE, Karanja DM, Secor WE: Evaluation of urine CCA assays for detection of Schistosoma mansoni infection in Western Kenya. PLoS Negl Trop Dis 2011, 5:e951.
• [19]Coulibaly JT, Knopp S, N'Guessan NA, Silue KD, Furst T, Lohourignon LK, Brou JK, N'Gbesso YK, Vounatsou P, N'Goran EK, Utzinger J: Accuracy of urine circulating cathodic antigen (CCA) test for Schistosoma mansoni diagnosis in different settings of Cote d'Ivoire. PLoS Negl Trop Dis 2011, 5:e1384.
• [20]Qian CY, Huang B, Yu CX, Zhang J, Yin XR, Wang J, Song LJ, Zhang W, Ke XD: Detection of the circulating antigen 14-3-3 protein of Schistosoma japonicum by time-resolved fluoroimmunoassay in rabbits. Parasit Vectors 2011, 4:95. BioMed Central Full Text
• [21]Vermeer HJ, van Dam GJ, Halkes KM, Kamerling JP, Vliegenthart JF, Hokke CH, Deelder AM: Immunodiagnostically applicable monoclonal antibodies to the circulating anodic antigen of Schistosoma mansoni bind to small, defined oligosaccharide epitopes. Parasitol Res 2003, 90:330-336.
• [22]Polman K, De Vlas SJ, Van Lieshout L, Deelder AM, Gryseels B: Evaluation of density-dependent fecundity in human Schistosoma mansoni infections by relating egg counts to circulating antigens through Deming regression. Parasitology 2001, 122:161-167.
• [23]Polman K, Diakhate MM, Engels D, Nahimana S, Van Dam GJ, Falcao Ferreira ST, Deelder AM, Gryseels B: Specificity of circulating antigen detection for schistosomiasis mansoni in Senegal and Burundi. Trop Med Int Health 2000, 5:534-537.
• [24]Al-Sherbiny MM, Osman AM, Hancock K, Deelder AM, Tsang VC: Application of immunodiagnostic assays: detection of antibodies and circulating antigens in human schistosomiasis and correlation with clinical findings. AmJTrop Med Hyg 1999, 60:960-966.
• [25]De Clercq D, Sacko M, Vercruysse J, vanden Bussche V, Landoure A, Diarra A, Gryseels B, Deelder A: Circulating anodic and cathodic antigen in serum and urine of mixed Schistosoma haematobium and S. mansoni infections in Office du Niger, Mali. Trop Med Int Health 1997, 2:680-685.
• [26]Deelder AM, van Dam GJ, Kornelis D, Fillie YE, van Zeyl RJ: Schistosoma: analysis of monoclonal antibodies reactive with the circulating antigens CAA and CCA. Parasitology 1996, 112(Pt 1):21-35.
• [27]Standley C, Lwambo N, Lange C, Kariuki H, Adriko M, Stothard J: Performance of circulating cathodic antigen (CCA) urine-dipsticks for rapid detection of intestinal schistosomiasis in schoolchildren from shoreline communities of Lake Victoria. Parasit Vectors 2010, 3:7. BioMed Central Full Text
• [28]Tini M, Jewell UR, Camenisch G, Chilov D, Gassmann M: Generation and application of chicken egg-yolk antibodies. Comp Biochem Physiol A Mol Integr Physiol 2002, 131:569-574.
• [29]Larsson A, Wejaker PE, Forsberg PO, Lindahl T: Chicken antibodies: a tool to avoid interference by complement activation in ELISA. J Immunol Methods 1992, 156:79-83.
• [30]Larsson A, Karlsson-Parra A, Sjoquist J: Use of chicken antibodies in enzyme immunoassays to avoid interference by rheumatoid factors. Clin Chem 1991, 37:411-414.
• [31]Carlander D, Kollberg H, Wejaker PE, Larsson A: Peroral immunotherapy with yolk antibodies for the prevention and treatment of enteric infections. Immunol Res 2000, 21:1-6.
• [32]Carlander D, Stalberg J, Larsson A: Chicken antibodies: a clinical chemistry perspective. Ups J Med Sci 1999, 104:179-189.
• [33]Shin SJ, Lee SS, Manning EJ, Collins MT: Production of and applications for a polyclonal IgY diagnostic reagent specific for Mycobacterium avium subsp. paratuberculosis. J Microbiol 2009, 47:600-609.
• [34]Empey Campora C, Hokama Y, Yabusaki K, Isobe M: Development of an enzyme-linked immunosorbent assay for the detection of ciguatoxin in fish tissue using chicken immunoglobulin Y. J Clin Lab Anal 2008, 22:239-245.
• [35]Juliarena M, Gutierrez S, Ceriani C: Chicken antibodies: a useful tool for antigen capture ELISA to detect bovine leukaemia virus without cross-reaction with other mammalian antibodies. Vet Res Commun 2007, 31:43-51.
• [36]Lei JH, Liu WQ, Sun CS, Tang CL, Li MJ, Chen YL, Li YL: Detection of circulating antigen in serum of mice infected with Schistosoma japonicum by immunomagnetic bead ELISA based on IgY. Acta Trop 2009, 111:39-43.
• [37]Lei JH, Su BT, Xu H, Shen JL, Guan XH, Feng ZQ, Li YL, Xu MX, Liu WQ: Evaluation of an IgY-based immunomagnetic enzyme-linked immunosorbent assay system for detection of circulating Schistosoma japonicum antigen in serum samples from patients in China. AmJTrop Med Hyg 2011, 85:1054-1059.
• [38]Liu F, Cui SJ, Hu W, Feng Z, Wang ZQ, Han ZG: Excretory/secretory proteome of the adult developmental stage of human blood fluke, Schistosoma japonicum. Mol Cell Proteomics 2009, 8:1236-1251.
• [39]Li YS, Ross AG, Sleigh AC, Li Y, Waine GJ, Williams GJ, Tanner M, McManus DP: Antibody isotype responses, infection and re-infection for Schistosoma japonicum in a marshland area of China. Acta Trop 1999, 73:79-92.
• [40]Aban JL, Ramajo V, Arellano JL, Oleaga A, Hillyer GV, Muro A: A fatty acid binding protein from Fasciola hepatica induced protection in C57/BL mice from challenge infection with Schistosoma bovis. Vet Parasitol 1999, 83:107-121.
• [41]Liu S, Dong W, Kong T: Preparation and characterization of immunoglobulin yolk against the venom of Naja naja atra. Indian J Exp Biol 2010, 48:778-785.
• [42]Ruan GP, Ma L, He XW, Meng MJ, Zhu Y, Zhou MQ, Hu ZM, Wang XN: Efficient production, purification, and application of egg yolk antibodies against human HLA-A*0201 heavy chain and light chain (beta2m). Protein Expr Purif 2005, 44:45-51.
• [43]Ko KY, Ahn DU: Preparation of immunoglobulin Y from egg yolk using ammonium sulfate precipitation and ion exchange chromatography. Poult Sci 2007, 86:400-407.
• [44]de Moraes MH, Guarneri AA, Girardi FP, Rodrigues JB, Eger I, Tyler KM, Steindel M, Grisard EC: Different serological cross-reactivity of Trypanosoma rangeli forms in Trypanosoma cruzi-infected patients sera. Parasit Vectors 2008, 1:20. BioMed Central Full Text
• [45]Johnston KL, Wu B, Guimaraes A, Ford L, Slatko BE, Taylor MJ: Lipoprotein biosynthesis as a target for anti-Wolbachia treatment of filarial nematodes. Parasit Vectors 2010, 3:99. BioMed Central Full Text
• [46]Liu F, Lu J, Hu W, Wang SY, Cui SJ, Chi M, Yan Q, Wang XR, Song HD, Xu XN, Wang JJ, Zhang XL, Zhang X, Wang ZQ, Xue CL, Brindley PJ, McManus DP, Yang PY, Feng Z, Chen Z, Han ZG: New perspectives on host-parasite interplay by comparative transcriptomic and proteomic analyses of Schistosoma japonicum. PLoS Pathog 2006, 2:e29.
• [47]Liu F, Hu W, Cui SJ, Chi M, Fang CY, Wang ZQ, Yang PY, Han ZG: Insight into the host-parasite interplay by proteomic study of host proteins copurified with the human parasite, Schistosoma japonicum. Proteomics 2007, 7:450-462.
• [48]Van Dam GJ, Bergwerff AA, Thomas-Oates JE, Rotmans JP, Kamerling JP, Vliegenthart JF, Deelder AM: The immunologically reactive O-linked polysaccharide chains derived from circulating cathodic antigen isolated from the human blood fluke Schistosoma mansoni have Lewis x as repeating unit. Eur J Biochem 1994, 225:467-482.
• [49]Bergwerff AA, van Dam GJ, Rotmans JP, Deelder AM, Kamerling JP, Vliegenthart JF: The immunologically reactive part of immunopurified circulating anodic antigen from Schistosoma mansoni is a threonine-linked polysaccharide consisting of – > 6)-(beta-D-GlcpA-(1 – > 3))-beta-D-GalpNAc-(1 – > repeating units. J Biol Chem 1994, 269:31510-31517.
• [50]Masciadri B, Areces LB, Carpinelli P, Foiani M, Draetta G, Fiore F: Characterization of the BUD31 gene of Saccharomyces cerevisiae. Biochem Biophys Res Commun 2004, 320:1342-1350.
• [51]Saha D, Khandelia P, O'Keefe RT, Vijayraghavan U: Saccharomyces cerevisiae NineTeen complex (NTC)-associated factor Bud31/Ycr063w assembles on precatalytic spliceosomes and improves first and second step pre-mRNA splicing efficiency. J Biol Chem 2012, 287:5390-5399.
• [52]Dunne DW, Jones FM, Doenhoff MJ: The purification, characterization, serological activity and hepatotoxic properties of two cationic glycoproteins (alpha 1 and omega 1) from Schistosoma mansoni eggs. Parasitology 1991, 103(Pt 2):225-236.
• [53]Fitzsimmons CM, Schramm G, Jones FM, Chalmers IW, Hoffmann KF, Grevelding CG, Wuhrer M, Hokke CH, Haas H, Doenhoff MJ, Dunne DW: Molecular characterization of omega-1: a hepatotoxic ribonuclease from Schistosoma mansoni eggs. Mol Biochem Parasitol 2005, 144:123-127.
• [54]Everts B, Perona-Wright G, Smits HH, Hokke CH, van der Ham AJ, Fitzsimmons CM, Doenhoff MJ, van der Bosch J, Mohrs K, Haas H, Mohrs M, Yazdanbakhsh M, Schramm G: Omega-1, a glycoprotein secreted by Schistosoma mansoni eggs, drives Th2 responses. J Exp Med 2009, 206:1673-1680.
• [55]Steinfelder S, Andersen JF, Cannons JL, Feng CG, Joshi M, Dwyer D, Caspar P, Schwartzberg PL, Sher A, Jankovic D: The major component in schistosome eggs responsible for conditioning dendritic cells for Th2 polarization is a T2 ribonuclease (omega-1). J Exp Med 2009, 206:1681-1690.
• [56]Chaudhuri J, Martin RW, Donahue MJ: Tryptophan hydroxylase and aromatic L-amino acid decarboxylase activities in the tissues of adult Ascaris suum. Int J Parasitol 1988, 18:341-346.
• [57]Cass CL, Johnson JR, Califf LL, Xu T, Hernandez HJ, Stadecker MJ, Yates JR, Williams DL: Proteomic analysis of Schistosoma mansoni egg secretions. Mol Biochem Parasitol 2007, 155:84-93.
• [58]Braschi S, Borges WC, Wilson RA: Proteomic analysis of the schistosome tegument and its surface membranes. Mem Inst Oswaldo Cruz 2006, 101(Suppl 1):205-212.
• [59]Mulvenna J, Moertel L, Jones MK, Nawaratna S, Lovas EM, Gobert GN, Colgrave M, Jones A, Loukas A, McManus DP: Exposed proteins of the Schistosoma japonicum tegument. Int J Parasitol 2010, 40:543-554.
• [60]Delcroix M, Medzihradsky K, Caffrey CR, Fetter RD, McKerrow JH: Proteomic analysis of adult S. mansoni gut contents. Mol Biochem Parasitol 2007, 154:95-97.