A common oocyst surface antigen of Cryptosporidium recognized by monoclonal antibodies

J. R. Yu; S. O'Hara; J. Lin; M. Dailey; G. Cain; J. L.C. Lin

doi:10.1007/s00436-001-0586-z

A common oocyst surface antigen of Cryptosporidium recognized by monoclonal antibodies

J. R. Yu, S. O'Hara, J. Lin, M. Dailey, G. Cain, J. L.C. Lin

Gastroenterology and Hepatology

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

Two hybridoma clones, CMYL3 and CMYL30, were generated by immunizing Balb/c mice with excysted oocysts of Cryptosporidium muris. Both clones secreted monoclonal antibodies against an oocyst-wall antigen with apparent molecular mass of 250 kDa (called CM250) from C. muris and C. parvum. The epitope appeared to be periodate-sensitive, suggesting the involvement of a carbohydrate moiety. Immunofluorescence and confocal microscopy on purified oocysts and infected mouse tissues revealed staining confined to the oocyst wall of both Cryptosporidium species. Immunogold labeling further revealed the presence of the CM250 antigen in electron-dense vesicles and cytoplasm of developing macrogametocytes, and ultimately localized to the oocyst wall of mature oocysts. Both antibodies cross-reacted with C. serpentis oocysts but did not recognize the other enteropathogenic protozoans Giardia muris, Eimeria falciformis and E. nischulz. These antibodies may be valuable tools for the analysis of oocyst-wall formation in Cryptosporidium and characterization of the common antigen.

Original language	English (US)
Pages (from-to)	412-420
Number of pages	9
Journal	Parasitology Research
Volume	88
Issue number	5
DOIs	https://doi.org/10.1007/s00436-001-0586-z
State	Published - 2002

ASJC Scopus subject areas

Parasitology
General Veterinary
Insect Science
Infectious Diseases

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title = "A common oocyst surface antigen of Cryptosporidium recognized by monoclonal antibodies",

abstract = "Two hybridoma clones, CMYL3 and CMYL30, were generated by immunizing Balb/c mice with excysted oocysts of Cryptosporidium muris. Both clones secreted monoclonal antibodies against an oocyst-wall antigen with apparent molecular mass of 250 kDa (called CM250) from C. muris and C. parvum. The epitope appeared to be periodate-sensitive, suggesting the involvement of a carbohydrate moiety. Immunofluorescence and confocal microscopy on purified oocysts and infected mouse tissues revealed staining confined to the oocyst wall of both Cryptosporidium species. Immunogold labeling further revealed the presence of the CM250 antigen in electron-dense vesicles and cytoplasm of developing macrogametocytes, and ultimately localized to the oocyst wall of mature oocysts. Both antibodies cross-reacted with C. serpentis oocysts but did not recognize the other enteropathogenic protozoans Giardia muris, Eimeria falciformis and E. nischulz. These antibodies may be valuable tools for the analysis of oocyst-wall formation in Cryptosporidium and characterization of the common antigen.",

author = "Yu, {J. R.} and S. O'Hara and J. Lin and M. Dailey and G. Cain and Lin, {J. L.C.}",

note = "Funding Information: Acknowledgements We would like to thank Dr. Donald W. Duszynski for the generous gift of the oocysts of Eimeria falciformis and E. nieschulzi, and Dr. Stanley L. Erlandsen for the cysts of Giardia muris. We also thank Rebecca Reiter for critical reading of the manuscript. This study was partly supported by grant no. 2000-1-20200-003-1 from the Basic Research Program of the Korean Science and Engineering Foundation (to J.R.Y.) and by grant HD18577 from the National Institutes of Health (to J.J.C.L.). The experiments comply with the current laws of the United States in which the experiments were performed.",

year = "2002",

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T1 - A common oocyst surface antigen of Cryptosporidium recognized by monoclonal antibodies

AU - Yu, J. R.

AU - O'Hara, S.

AU - Lin, J.

AU - Dailey, M.

AU - Cain, G.

AU - Lin, J. L.C.

N1 - Funding Information: Acknowledgements We would like to thank Dr. Donald W. Duszynski for the generous gift of the oocysts of Eimeria falciformis and E. nieschulzi, and Dr. Stanley L. Erlandsen for the cysts of Giardia muris. We also thank Rebecca Reiter for critical reading of the manuscript. This study was partly supported by grant no. 2000-1-20200-003-1 from the Basic Research Program of the Korean Science and Engineering Foundation (to J.R.Y.) and by grant HD18577 from the National Institutes of Health (to J.J.C.L.). The experiments comply with the current laws of the United States in which the experiments were performed.

PY - 2002

Y1 - 2002

N2 - Two hybridoma clones, CMYL3 and CMYL30, were generated by immunizing Balb/c mice with excysted oocysts of Cryptosporidium muris. Both clones secreted monoclonal antibodies against an oocyst-wall antigen with apparent molecular mass of 250 kDa (called CM250) from C. muris and C. parvum. The epitope appeared to be periodate-sensitive, suggesting the involvement of a carbohydrate moiety. Immunofluorescence and confocal microscopy on purified oocysts and infected mouse tissues revealed staining confined to the oocyst wall of both Cryptosporidium species. Immunogold labeling further revealed the presence of the CM250 antigen in electron-dense vesicles and cytoplasm of developing macrogametocytes, and ultimately localized to the oocyst wall of mature oocysts. Both antibodies cross-reacted with C. serpentis oocysts but did not recognize the other enteropathogenic protozoans Giardia muris, Eimeria falciformis and E. nischulz. These antibodies may be valuable tools for the analysis of oocyst-wall formation in Cryptosporidium and characterization of the common antigen.

AB - Two hybridoma clones, CMYL3 and CMYL30, were generated by immunizing Balb/c mice with excysted oocysts of Cryptosporidium muris. Both clones secreted monoclonal antibodies against an oocyst-wall antigen with apparent molecular mass of 250 kDa (called CM250) from C. muris and C. parvum. The epitope appeared to be periodate-sensitive, suggesting the involvement of a carbohydrate moiety. Immunofluorescence and confocal microscopy on purified oocysts and infected mouse tissues revealed staining confined to the oocyst wall of both Cryptosporidium species. Immunogold labeling further revealed the presence of the CM250 antigen in electron-dense vesicles and cytoplasm of developing macrogametocytes, and ultimately localized to the oocyst wall of mature oocysts. Both antibodies cross-reacted with C. serpentis oocysts but did not recognize the other enteropathogenic protozoans Giardia muris, Eimeria falciformis and E. nischulz. These antibodies may be valuable tools for the analysis of oocyst-wall formation in Cryptosporidium and characterization of the common antigen.

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A common oocyst surface antigen of Cryptosporidium recognized by monoclonal antibodies

Abstract

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