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Infection and Immunity, July 2002, p. 3881-3890, Vol. 70, No. 7
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.7.3881-3890.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Analysis of Sequence Diversity at the Highly Polymorphic Cpgp40/15 Locus among Cryptosporidium Isolates from Human Immunodeficiency Virus-Infected Children in South Africa

Brett A. Leav,1 Malanie R. Mackay,1,2 Akudo Anyanwu,1 Roberta M. O' Connor,1 Ana Maria Cevallos,1,{dagger} Gurpreet Kindra,3 Nigel C. Rollins,2,3 Michael L. Bennish,1,2 Richard G. Nelson,4 and Honorine D. Ward1*

Division of Geographic Medicine and Infectious Diseases, Tufts New England Medical Center, Boston, Massachusetts,1 Nelson R. Mandela School of Medicine, University of Natal, Durban,2 Africa Center for Health and Population Studies, Mtubatuba, South Africa,3 Division of Infectious Diseases, San Francisco General Hospital, University of California, San Francisco, California4

Received 10 December 2001/ Returned for modification 2 February 2002/ Accepted 6 April 2002

Cryptosporidium sp. is a significant cause of diarrheal disease, particularly in human immunodeficiency virus (HIV)-infected patients in developing countries. We recently cloned and sequenced several alleles of the highly polymorphic single-copy Cryptosporidium parvum gene Cpgp40/15. This gene encodes a precursor protein that is proteolytically cleaved to yield mature cell surface glycoproteins gp40 and gp15, which are implicated in zoite attachment to and invasion of enterocytes. The most-striking feature of the Cpgp40/15 alleles and proteins is their unprecedented degree of sequence polymorphism, which is far greater than that observed for any other gene or protein studied in C. parvum to date. In this study we analyzed nucleic acid and amino acid sequence polymorphism at the Cpgp40/15 locus of 20 C. parvum isolates from HIV-infected South African children. Fifteen isolates exhibited one of four previously identified genotype I alleles at the Cpgp40/15 locus (Ia, Ib, Ic, and Id), while five displayed a novel set of polymorphisms that defined a new Cpgp40/15 genotype I allele, designated genotype Ie. Surprisingly, only 15 of these isolates exhibited concordant type I alleles at the thrombospondin-related adhesive protein of Cryptosporidium and Cryptosporidium oocyst wall protein loci, while five isolates (all of which displayed Cpgp40/15 genotype Ic alleles) displayed genotype II alleles at these loci. Furthermore, the last five isolates also manifested chimeric genotype Ic/Ib or Ic/II alleles at the Cpgp40/15 locus, raising the possibility of sexual recombination within and between prototypal parasite genotypes. Lastly, children infected with isolates having genotype Ic alleles were significantly older than those infected with isolates displaying other genotype I alleles.

* Corresponding author. Mailing address: Division of Geographic Medicine and Infectious Diseases, New England Medical Center, Box 04, 750 Washington St., Boston, MA 02111. Phone: (617) 636-7032. Fax: (617) 636-5292. E-mail: Hward{at}lifespan.org.

Editor: W. A. Petri, Jr.

{dagger} Present address: Departamento de Biología Molecular, Instituto de Investigaciones Biomédicas, UNAM, Ciudad Universitaria 04510, Mexico.

Infection and Immunity, July 2002, p. 3881-3890, Vol. 70, No. 7
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.7.3881-3890.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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