Proteome approach for identification of schistosomiasis japonica vaccine candidate antigen. Proteome approach for identification of schistosomiasis japonica vaccine candidate antigen

Access this Article

Search this Article

Author(s)

    • KIKUCHI Mihoko
    • Department of Immunogenetics, Institute of Tropical Medicine (NEKKEN), Nagasaki University
    • WATANABE Kanji
    • Department of Parasitology, Institute of Tropical Medicine (NEKKEN), Nagasaki University
    • NARA Takeshi
    • Department of Molecular and Cellular Parasitology, Department of Epidemiology and Environmental Health
    • ARAKAWA Takeshi
    • Division of Molecular Microbiology, Center of Molecular Biosciences, University of the Ryukyu
    • AOKI Yoshiki
    • Department of Parasitology, Institute of Tropical Medicine (NEKKEN), Nagasaki University
    • HIRAYAMA Kenji
    • Department of Immunogenetics, Institute of Tropical Medicine (NEKKEN), Nagasaki University

Abstract

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.

Journal

  • Parasitology international

    Parasitology international 58(1), 36-44, 2009-03-01

    ELSEVIER

References:  53

Codes

  • NII Article ID (NAID)
    10025329984
  • NII NACSIS-CAT ID (NCID)
    AA11112001
  • Text Lang
    ENG
  • Article Type
    ART
  • ISSN
    13835769
  • Data Source
    CJP  IR 
Page Top