Abstract
Cryptosporidium parvum is a zoonotic intracellular protozoan responsible for the diarrheal illness cryptosporidiosis in humans and animals. Although a number of zoite surface proteins are known to be expressed during, and believed to be involved in, attachment and invasion of host cells, the molecular mechanisms by which C. parvum invades the host epithelial cells are not well understood. In the present study, we investigated the gene expression patterns, protein localization in developmental stages in culture, and in vitro neutralization characteristics of Cpgp40/15 and Cpgp40. Indirect immunofluorescence assay showed that Cpgp40/15 is associated with the parasitophorous vacuole membrane (PVM) during intracellular development. Both anti-gp40/15 and anti-gp40 antibodies demonstrated the ability to neutralize C. parvum infection in vitro. Further studies are needed to fully understand the specific role and functional mechanism of Cpgp40/15 (or gp40/15 complex) in the invasion of the host or in the PVM and to determine the feasibility of gp40/15 as a vaccine candidate for cryptosporidiosis in vivo.
Highlights
Cryptosporidium spp. are important etiological agents of diarrhea and are among the leading causes of moderate to severe diarrhea in children under 2 years [1]
The host cell-derived parasitophorous vacuole membrane (PVM) structure separates the intracellular parasites from the host cell cytosol [12]
We investigated the gene expression patterns, protein localization in developmental stages in culture, and in vitro neutralization characteristics of Cpgp40/15 and Cpgp40 to gain deeper insights into the biological role of Cpgp40/15 in C. parvum
Summary
Cryptosporidium spp. are important etiological agents of diarrhea and are among the leading causes of moderate to severe diarrhea in children under 2 years [1]. Cryptosporidiosis is self-limiting in immunocompetent hosts but can be a chronic and life-threatening infection in immunocompromised patients [2]. Nitazoxanide is approved for treatment of cryptosporidiosis in immunocompetent individuals, it has not been approved for use by immunocompromised patients [5]. The initial interaction processes of Cryptosporidium oocysts and sporozoites with host epithelial cells can be divided into several major developmental phases: excystation, gliding motility, attachment, invasion, parasitophorous vacuole formation, intracellular maintenance, and host cell damage [9,10]. Cryptosporidium does not normally cause systemic infection or penetrate deep tissue; rather, the parasite establishes itself in a membrane-bound compartment, termed the parasitophorous vacuole (PV), on the apical surface of the intestinal epithelium [11]. The host cell-derived parasitophorous vacuole membrane (PVM) structure separates the intracellular parasites from the host cell cytosol [12]
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