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Infection and Immunity, July 2000, p. 4117-4134, Vol. 68, No. 7
Division of Infectious Diseases, San
Francisco General Hospital,1 and
Departments of Medicine2 and
Pharmaceutical Chemistry,3 University of
California, San Francisco, San Francisco, California 94143-0811
Received 22 February 2000/Accepted 1 April 2000
The apicomplexan parasite Cryptosporidium parvum is a
major cause of serious diarrheal disease in both humans and animals. No
efficacious chemo- or immunotherapies have been identified for
cryptosporidiosis, but certain antibodies directed against zoite
surface antigens and/or proteins shed by gliding zoites have been shown
to neutralize infectivity in vitro and/or to passively protect against,
or ameliorate, disease in vivo. We previously used monoclonal antibody
11A5 to identify a 15-kDa surface glycoprotein that was
shed behind motile sporozoites and was recognized by several lectins
that neutralized parasite infectivity for cultured epithelial cells.
Here we report the cloning and sequence analysis of the gene encoding
this 11A5 antigen. Surprisingly, the gene encoded a 330-amino-acid,
mucin-like glycoprotein that was predicted to contain an
N-terminal signal peptide, a homopolymeric tract of serine residues, 36 sites of O-linked glycosylation, and a hydrophobic C-terminal peptide
specifying attachment of a glycosylphosphatidylinositol anchor. The
single-copy gene lacked introns and was expressed during merogony to
produce a 60-kDa precursor which was proteolytically cleaved to 15- and
45-kDa glycoprotein products that both localized to the
surface of sporozoites and merozoites. The gp15/45/60 gene displayed a
very high degree of sequence diversity among C. parvum isolates, and the numerous single-nucleotide and single-amino-acid polymorphisms defined five to six allelic classes, each characterized by additional intra-allelic sequence variation. The gp15/45/60 single-nucleotide polymorphisms will prove useful for haplotyping and
fingerprinting isolates and for establishing meaningful relationships between C. parvum genotype and phenotype.
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Cloning and Sequence Analysis of a Highly
Polymorphic Cryptosporidium parvum Gene Encoding a
60-Kilodalton Glycoprotein and Characterization of Its 15- and
45-Kilodalton Zoite Surface Antigen Products
*
Corresponding author. Mailing address: Box 0811, University of California, San Francisco, CA 94143-0811. Phone: (415)
206-8846. Fax: (415) 648-8425. E-mail:
malaria{at}itsa.ucsf.edu.
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