Proteome approach for identification of schistosomiasis japonica vaccine candidate antigen.

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概要

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• Experimental vaccination with radiation-attenuated cercariae (RAC) confers possible practical levels of resistance to challenge infection by humoral and by cellular mechanism. Here, we aimed to identify possible vaccine antigens by using specific IgG antibody from RAC vaccinated miniature pig. Two milligrams of soluble egg antigen (SEA) or schistosomal worm antigen preparation (SWAP) was fractionated using two dimensional liquid chromatography (proteome PF 2D) consisted of high performance chromatofocusing (HPCF) and high resolution reversed phase chromatography (HPRP). Of the 42 HPCF fractions of SEA or SWAP, 26 (61.9%) or 15 (35.7%) showed positive dot blot reaction with RAC vaccinated serum respectively. The dot blot positive fractions were applied to the second HPRP column. One hundred and seven out of 26 x 96 of SEA fractions and 18 out of 15 x 96 SWAP fractions reacted with RAC vaccinated serum. From the positive fractions we chose 17 of SEA and 10 of SWAP that had no reactivity with normal cercariae infected (NCI) sera and had single peak of 214 nm; and automated N-terminal amino acid sequence based on in situ Edman Reaction was conducted. Four sequences were obtained and applied to the homology search in NCBI database. A total of eight candidate genes were listed up and their cDNA clones from schistosomula stage were obtained. Two of the recombinant proteins (AAW27472.1 and AXX25883.1) showed strong reactivity with the RAC vaccinated serum but marginal with NCI serum. This protocol using proteome PF 2D could be applicable in identifying immunoreactive proteins from crude extract for the development of vaccines or for diagnostics.

• 2009-03-01

著者

• HIRAYAMA Kenji

  Department of Physics, Chiba University

• Ito Takashi

  Department Of Physics University Of Tokyo

• Aoki Yoshiki

  Department of Parasitology, Institute of Tropical Medicine, Nagasaki University

• Kikuchi Mihoko

  Department Of Immunogenetics Institute Of Tropical Medicine (nekken) Nagasaki University

• Hirayama Kenji

  Department Of Immunogenetics Institute Of Tropical Medicine (nekken) Nagasaki University

• ABDEL-HAFEEZ Ekhlas

  Department of Immunogenetics, Institute of Tropical Medicine (NEKKEN), Nagasaki University

• WATANABE Kanji

  Department of Parasitology, Institute of Tropical Medicine (NEKKEN), Nagasaki University

• YU Chuanxin

  Jiangsu Institute of Parasitic Diseases

• CHEN Honggen

  Jiangxi Provincial Institute of Parasitic Diseases

• NARA Takeshi

  Department of Molecular and Cellular Parasitology, Department of Epidemiology and Environmental Heal

• ARAKAWA Takeshi

  Division of Molecular Microbiology, Center of Molecular Biosciences, University of the Ryukyu

• Hirayama Kenji

  Department Of Clinical Immunology Medical Institute Of Bioregulation Kyushu University

• Watanabe Kanji

  Department Of Clinical Nutrition Faculty Of Comprehensive Rehabilitation Osaka Prefecture University

• Abdel-hafeez Ekhlas

  Department Of Immunogenetics Institute Of Tropical Medicine (nekken) Nagasaki University

• Arakawa Takeshi

  Division Of Molecular Microbiology Center Of Molecular Biosciences University Of The Ryukyu

• Ito Takashi

  Department Of Chemistry School Of Science The University Of Tokyo

• Nara Takeshi

  Department Of Molecular And Cellular Parasitology Department Of Epidemiology And Environmental Healt

• Aoki Yoshiki

  Department Of Parasitology Institute Of Tropical Medicine (nekken) Nagasaki University

• Aoki Yoshiki

  Department Of Parasitology Institute For Tropical Medicine Nagasaki University

• Watanabe Kanji

  Department Of Parasitology Institute Of Tropical Medicine (nekken) Nagasaki University

• ITO Takashi

  Department of Advanced Materials Chemistry, Graduate School of Engineering, Yokohama National University

• ITO Takashi

  Department of Biochemistry, Nagasaki University

• KIKUCHI Mihoko

  Department of Immunogenetics and Global Centre of Excellence, Institute of Tropical Medicine, Nagasaki University

• AOKI Yoshiki

  Department of Agricultural Chemistry, Gifu University

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