title: Selection and identification of a new adhesion protein of Cryptosporidium parvum from a cDNA library by ribosome display.
abstract: In this study, we described a novel display method to identify surface adhesion proteins of Cryptosporidium parvum. A cDNA library of the sporozoite and oocyst stages of C. parvum was expressed on ribosome and selectively and specifically screened with intestinal epithelial cells (IECs) from newborn Cryptosporidium-free Holstein calves. Proteins were then enriched using a multi-step panning procedure. A new surface adherence protein of C. parvum was selected, named Cp20. Sequence analyses showed that Cp20 has a N-terminal signal peptide and four transmembrane regions. Indirect immunofluorescence assay (IFA) using an antibody specific for rCp20 demonstrated that the antibody specifically bound to the surface of sporozoites and oocysts. The recombinant plasmid pVAX1-Cp20 was constructed to examine the potential of the Cp20 gene as a target for specific preventive and therapeutic measures for cryptosporidiosis. The in vivo efficacies of the DNA vaccine was tested in BALB/c mice. The results indicated that the DNA vaccine elicited significant antibody responses and specific cellular responses when compared to control mice that received vector only or PBS. The DNA vaccine induced strong protective immune response against C. parvum and lower level of the oocysts shedding after challenge infection. This study suggested that Cp20 could serve as an effective target for specific preventive and therapeutic measures for cryptosporidiosis.
authors: College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: Dan
authors: D
authors: Xiao
authors: Chi
authors: C
authors: Yin
authors: Qian
authors: Q
authors: Zhang
authors: Jian-hua
authors: JH
authors: Li
authors: Peng-tao
authors: PT
authors: Gong
authors: Shu-hong
authors: SH
authors: Li
authors: Guo-cai
authors: GC
authors: Zhang
authors: Ying-jie
authors: YJ
authors: Gao
authors: Xi-chen
authors: XC
authors: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D015815:Cell Adhesion Molecules / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D003457:Cryptosporidiosis / Q000276:immunology / Q000469:parasitology / Q000517:prevention & control; D016785:Cryptosporidium parvum / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D015723:Gene Library; D006367:HeLa Cells; D006790:Host-Parasite Interactions; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000469:parasitology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010957:Plasmids / Q000276:immunology; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012270:Ribosomes / Q000235:genetics*; D014162:Transfection; D019444:Vaccines, DNA / Q000276:immunology
chemical_list: D015815:Cell Adhesion Molecules; D015800:Protozoan Proteins; D012333:RNA, Messenger; D019444:Vaccines, DNA