Preparation and characterization of monoclonal antibodies against ES antigens of Trichinella spiralis

Cui, J.; Jing, F.; Fu, G.; Ren, H.; Liu, L.; Wang, Z.

doi:10.2478/s11687-014-0238-3

Abstract

Hybridomas secreting monoclonal antibodies (MAbs) against excretory-secretory (ES) antigen of Trichinella spiralis muscle larvae were produced. The 12 monoclones (designated 3A11, 7D9, 7F6, 5A7, 6H7, 39F3, 38H2, 37E7, 35B9, 29A11, 39C9 and 39A6) secreted IgM, IgG3 and IgG2b, respectively. Western blot using T. spiralis ES antigens showed that ten of 12 MAbs recognized the bands between 119.8 − 19.3kDa (mainly 68.4 − 28.7kDa) and two MAbs (3A11, 37E7) did not recognize any protein constituents of ES antigens. Nine of 12 MAbs also recognized the larval antigens of T. nativa, T. pseudospiralis and T. nelsoni at 40 days post-infection (dpi). No cross-reactions were found with the somatic antigens of adult worms of P. westermani, C. sinensis, and T. solium cysticercus. However, three (3A11, 6H7 and 39A6) were cross-reacted with the somatic antigens of adult worms of S. japonicum. Immunofluorescent location showed that the nine MAbs reacted with the cuticle and internal organs of T. spiralis larvae. Additionally, five and eight MAbs generated in this study can recognize the early antigens of pre-encapsulated T. spiralis larvae at 11 dpi and 13 dpi, respectively. The generation and characterization of the MAbs against T. spiralis ES antigens provide foundation for the development of specific early serodiagnostic techniques for trichinellosis.

References

[1] Arriaga, C., Yepez-Mulia, L., Morilla, A., Ortegapierres, G. (1995): Detection of circulating Trichinella spiralis muscle larva antigens in serum samples of experimentally and naturally infected swine. Vet. Parasitol. 58: 319–326. DOI: 10.1016/0304-4017(94)00775-8 http://dx.doi.org/10.1016/0304-4017(94)00775-810.1016/0304-4017(94)00775-8
Search in Google Scholar
[2] Boireau, P., Vayssier, M., Fabien, J. F., Perret, C., Calamel, M., Soule, C. (1997): Characterization of eleven antigenic groups in Trichinella genus and identification of stage and species markers. Parasitology, 115: 641–651 http://dx.doi.org/10.1017/S003118209700171610.1017/S0031182097001716
Search in Google Scholar
[3] Bradford, M. M. (1976): A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72: 248–254. DOI: 10.1016/0003-2697(76)90527-3 http://dx.doi.org/10.1016/0003-2697(76)90527-310.1016/0003-2697(76)90527-3
Search in Google Scholar
[4] Bruschi, F., Tassi, C., Pozio, E. (1990): Parasite-specific antibody response in Trichinella sp. human infection: A one year follow-up. Am. J. Trop. Med. Hyg., 43: 186–193
Search in Google Scholar
[5] Chen, M. X., Zhang R. L, Chen, J. X., Chen, S. H., Li, X. H., Gao, S. T., Geng, Y. J., Huang, D. N., Ai, L., Xu, M. J., Zhu, X. Q. (2010): Monoclonal antibodies against excretory/secretory antigens of Angiostrongylus cantonensis. Hybridoma (Larchmt), 29: 447–452. DOI: 10.1089/hyb.2010.0008 http://dx.doi.org/10.1089/hyb.2010.000810.1089/hyb.2010.0008
Search in Google Scholar
[6] Cui, J., Wang, Z. Q., Zhu, W., Zhang, R. G. (1999): Seroepidemiological study of Trichinella spiralis infection in central China. Helminthologia, 36: 235–239
Search in Google Scholar
[7] Cui, J., Wang, Z. Q., Xu, B. L. (2011): The epidemiology of human trichinellosis in China during 2004–2009. Acta Trop., 118: 1–5. DOI: 10.1016/j.actatropica.2011.02.005 http://dx.doi.org/10.1016/j.actatropica.2011.02.00510.1016/j.actatropica.2011.02.005
Search in Google Scholar
[8] Dea-auuela, M. A., Bolas-Fernandez, F. (1999): Trichinella antigens: a review. Vet. Res., 30: 559–571
Search in Google Scholar
[9] De-La-Rosa, J. L., Moran-Tlatelpa, E., Medina, Y., Gomez-Priego, A., Correa, D. (2001): Detection of circulating and fecal Trichinella spiralis antigens during experimental infection using monoclonal antibodies against the new born larvae. Parasite, 8: S123–S125 http://dx.doi.org/10.1051/parasite/200108s212310.1051/parasite/200108s2123
Search in Google Scholar
[10] Gamble, H. R., Pozio, E., Bruschi, F., Nockler, K., Kapel, C. M., Gajadhar, A. A. (2004): International Commission on Trichinellosis: recommendations on the use of serological tests for the detection of Trichinella infection in animals and man. Parasite, 11: 3–13. DOI: 10.1051/parasite/20041113 http://dx.doi.org/10.1051/parasite/2004111310.1051/parasite/20041113
Search in Google Scholar
[11] Gamble, H. R., Rapic, D., Marinculic, A., Murrell, K. D. (1988): Evaluation of excretory-secretory antigens for the serodiagnosis of swine trichinellosis. Vet. Parasitol., 30: 131–137. DOI: 10.1016/0304-4017(88)90160-4 http://dx.doi.org/10.1016/0304-4017(88)90160-410.1016/0304-4017(88)90160-4
Search in Google Scholar
[12] Harnett, W., Macdonal, M., Preece, G., Patterson, M., Parkhouse, R.M. (1997): Production of monoclonal antibodies against excretory-secretory products of adult male Onchocerca gibsoni. J. Parasitol., 83: 316–319. DOI: 10.2307/3284464 http://dx.doi.org/10.2307/328446410.2307/3284464
Search in Google Scholar
[13] Kapel, C. M., Gamble, H. R. (2000): Infectivity, persistence, and antibody response to domestic and sylvatic Trichinella spp. in experimentally infected pigs. Int. J. Parasitol., 30: 215–221. DOI: 10.1016/S0020-7519 (99)00202-7 http://dx.doi.org/10.1016/S0020-7519(99)00202-710.1016/S0020-7519(99)00202-7
Search in Google Scholar
[14] Kapel, C. M., Webster, P., Lind, P., Pozio, E., Henriksen, S. A., Murrell, K. D., Nansen, P. (1998): Trichinella spiralis, T. britovi, and T. nativa: infectivity, larval distribution in muscle, and antibody response after experimental infection of pigs. Parasitol. Res., 84: 264–271. DOI: 10.1007/s004360050393 http://dx.doi.org/10.1007/s00436005039310.1007/s004360050393
Search in Google Scholar
[15] Kennett, R. H., Mckearn, T. J., Bechtol, K. B. (1980): Monoclonal antibodies: hybridomas, a new dimension in biological analyses. Plenum Press, New York, NY http://dx.doi.org/10.1007/978-1-4615-7505-410.1007/978-1-4615-7505-4
Search in Google Scholar
[16] Kohler, G., Milstein, J. (1975): Continuous cultures of fused cells secreting antibody of predefined specificity. Nature, 256: 495–497. DOI: 10.1038/256495a0 http://dx.doi.org/10.1038/256495a010.1038/256495a0
Search in Google Scholar
[17] Li, F., Cui, J., Wang, Z. Q., Jiang, P. (2010): Sensitivity and Optimization of Artificial Digestion in the Inspection of Meat for Trichinella spiralis. Foodborne Pathog. Dis., 7: 879–885. DOI: 10.1089/fpd.2009.0445 http://dx.doi.org/10.1089/fpd.2009.044510.1089/fpd.2009.0445
Search in Google Scholar
[18] Li, C. K., Ko, R. C. (2001): The detection and occurrence of circulating antigens of Trichinella spiralis during worm development. Parasitol. Res., 87: 155–162. DOI: 10.1007/PL00008569 http://dx.doi.org/10.1007/PL0000856910.1007/PL00008569
Search in Google Scholar
[19] Liu, L. N., Jing, F. J., Cui, J., Fu, G. Y., Wang, Z. Q. (2013): Detection of circulating antigen in serum of mice infected with Trichinella spiralis by an IgY-IgM mAb sandwich ELISA. Exp. Parasitol., 133: 150–155. DOI: 10.1007/PL00008569 http://dx.doi.org/10.1016/j.exppara.2012.11.00110.1007/PL00008569
Search in Google Scholar
[20] Mahannop, P., Chaicumpa, W., Setasuban, P., Morakote, N., Tapchaisri, P. (1992): Immunodiagnosis of human trichinellosis using excretory-secretory (ES) antigen. J. Helminthol., 66: 297–304. DOI: 10.1017/S0022149X00014759 http://dx.doi.org/10.1017/S0022149X0001475910.1017/S0022149X00014759
Search in Google Scholar
[21] Mckinney, M. M., Parkinson, A. (1987): A simple, nonchromatographic procedure to purify immunoglobulins from serum and ascites fluid. J. Immunol. Methods, 96: 271–278. DOI: 10.1016/0022-1759(87)90324-3 http://dx.doi.org/10.1016/0022-1759(87)90324-310.1016/0022-1759(87)90324-3
Search in Google Scholar
[22] Mclaren, D. J., Ortega-pierres, G., Parkhouse, R. M. (1987): Trichinella spiralis: immunocytochemical localization of surface and intracellular antigens using monoclonal antibody probes. Parasitology, 94: 101–114. DOI: 10.1017/S003118200005349X http://dx.doi.org/10.1017/S003118200005349X10.1017/S003118200005349X3547257
Search in Google Scholar
[23] NÖCKLER, K., Reckinger, S., Broglia, A., Mayerscholl, A., Bahn, P. (2009): Evaluation of a Western blot and ELISA for the detection of anti-Trichinella-IgG in pig sera. Vet. Parasitol., 163: 341–347. DOI: 10.1016/j.vetpar.2009.04.034 http://dx.doi.org/10.1016/j.vetpar.2009.04.03410.1016/j.vetpar.2009.04.03419473770
Search in Google Scholar
[24] Pozio, E. (2007): World distribution of Trichinella spp. infections in animals and humans. Vet. Parasitol., 149: 3–21. DOI: 10.1016/j.vetpar.2007.07.002 http://dx.doi.org/10.1016/j.vetpar.2007.07.00210.1016/j.vetpar.2007.07.002
Search in Google Scholar
[25] Romaris, F., Escalante, M., Valins, B., Seoane, R., Wang, Z. Q., Leiro, J., Ubeira, F. M. (2001): Characterization of two monoclonal antibodies raised in Btk(xid) mice that recognize phosphorylcholine-bearing antigens from Trichinella and other helminths. Parasite Immunol., 23: 313–322. DOI: 10.1046/j.1365-3024.2001.00388.x http://dx.doi.org/10.1046/j.1365-3024.2001.00388.x10.1046/j.1365-3024.2001.00388.x
Search in Google Scholar
[26] Srimanote, P., Ittiprasert, W., Sermsart, B., Chaisri, U., Mahannop, P., Sakolvaree, Y., Tapchaisri, P., Maleewong, W., Kurazono, H., Hayashi, H., Chaicumpa, W. (2000): Trichinella spiralis- specific monoclonal antibodies and affinity-purified antigen-based diagnosis. Asian Pac. J. Allergy Immunol., 18: 37–45
Search in Google Scholar
[27] Wang, L., Wang, Z. Q., Cui, J. (2013a): Proteomic analysis of the changed proteins of Trichinella spiralis infective larvae after co-culture in vitro with intestinal epithelial cells. Vet. Parasitol. 194: 160–163. DOI: 10.1016/j.vetpar.2013.01.045 http://dx.doi.org/10.1016/j.vetpar.2013.01.04510.1016/j.vetpar.2013.01.045
Search in Google Scholar
[28] Wang, L., Wang, Z. Q., Hu, D. D., Cui, J. (2013b): Proteomic analysis of Trichinella spiralis muscle larval excretory- secretory proteins recognized by early infection sera. Biomed. Res. Int., 2013: 139745. DOI: 10.1155/2013/139745 10.1155/2013/139745
Search in Google Scholar
[29] Wang, L., Cui, J., Hu, D. D., Liu, R. D., Wang, Z. Q. (2014): Identification of early diagnostic antigens from major excretory-secretory proteins of Trichinella spiralis muscle larvae using immunoproteomics. Parasit. Vectors, 7: 40. DOI: 10.1186/1756-3305-7-40 http://dx.doi.org/10.1186/1756-3305-7-4010.1186/1756-3305-7-40
Search in Google Scholar
[30] Wang, R., Wang, Z. Q., Cui, J. (2009): Immunodiagnostic value and immune protection of the recombinant Ts21 antigen of Trichinella spiralis. Chin. J. Parasitol. Parasit. Dis., 27: 17–21 (In Chinese)
Search in Google Scholar
[31] Wang, S. W., Wang, Z. Q., Cui, J. (2011): Protein change of intestinal epithelial cells induced in vitro by Trichinella spiralis infective larvae. Parasitol. Res., 108: 593–599. DOI: 10.1007/s00436-010-2102-9 http://dx.doi.org/10.1007/s00436-010-2102-910.1007/s00436-010-2102-9
Search in Google Scholar
[32] Wang, Z. Q., Cui, J., Wei, H. Y., Han, H. M., Zhang, H. W., Li, Y. L. (2006): Vaccination of mice with DNA vaccine induces the immune response and partial protection against T. spiralis infection. Vaccine, 24: 1205–1212. DOI: 10.1016/j.vaccine.2005.08.104 http://dx.doi.org/10.1016/j.vaccine.2005.08.10410.1016/j.vaccine.2005.08.104
Search in Google Scholar
[33] Wang, Z. Q., Cui, J., Wu, F., Mao, F. R., Jin, X. X. (1998): Epidemiological, clinical and serological studies on trichinellosis in Henan Province, China. Acta Trop., 71: 255–268. DOI: 10.1016/S0001-706X(98)00072-2 http://dx.doi.org/10.1016/S0001-706X(98)00072-210.1016/S0001-706X(98)00072-2
Search in Google Scholar
[34] Wang, Z. Q., Fu, G. Y., Jing F. J., Jin, J., Ren, H. J., Jiang, P., Cui, J. (2012a): Detection of Trichinella spiralis circulating antigens in serum of experimentally infected mice by an IgY-mAb sandwich ELISA. Foodborne Pathog. Dis., 9: 727–933. DOI: 10.1089/fpd.2012. 1157 http://dx.doi.org/10.1089/fpd.2012.1157
Search in Google Scholar
[35] Wang, Z. Q., Wang, L., Cui, J. (2012b): Proteomic analysis of Trichinella spiralis proteins in intestinal epithelial cells after culture with their larvae by shotgun LC-MS/MS approach. J. Proteomics, 75: 2375–2383. DOI: 10.1016/j.vetpar.2013.01.045 http://dx.doi.org/10.1016/j.jprot.2012.02.00510.1016/j.vetpar.2013.01.045
Search in Google Scholar
[36] Zhu, D. Z., Bell, R. G. (1990): Trichinella spiralis: murine strain variation in response to monoclonally defined, protective, nonstage-specific antigens. Exp. Parasitol., 70: 330–343. DOI: 10.1016/0014-4894(90)90115-S http://dx.doi.org/10.1016/0014-4894(90)90115-S10.1016/0014-4894(90)90115-S
Search in Google Scholar

Preparation and characterization of monoclonal antibodies against ES antigens of Trichinella spiralis

Abstract

Paradigm

My account