Premium
Molecular and immunological characterization of Babesia gibsoni and Babesia microti heat shock protein‐70
Author(s) -
TERKAWI M. ALAA,
ABOGE G.,
JIA H.,
GOO Y.K.,
OOKA H.,
YAMAGISHI J.,
NISHIKAWA Y.,
YOKOYAMA N.,
IGARASHI I.,
KAWAZU S.I.,
FUJISAKI K.,
XUAN X.
Publication year - 2009
Publication title -
parasite immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.795
H-Index - 75
eISSN - 1365-3024
pISSN - 0141-9838
DOI - 10.1111/j.1365-3024.2009.01109.x
Subject(s) - biology , immunoscreening , microbiology and biotechnology , recombinant dna , heat shock protein , virology , babesiosis , antibody , gene , adjuvant , open reading frame , peripheral blood mononuclear cell , complementary dna , babesia , in vitro , peptide sequence , immunology , cdna library , biochemistry
Summary Serological immunoscreening was used to identify a gene encoding heat shock protein‐70 from Babesia gibsoni (BgHSP‐70) that showed high homology with HSP‐70s from other apicomplexan parasites. This gene corresponded to a full‐length cDNA containing an open reading frame of 1968 bp predicted to result in a 70‐kDa mature protein consisting of 656 amino acids. Analysis of the expression levels of BgHSP‐70 indicated elevated transcription from cultured parasites incubated at 40°C for 1 h, but not at 30°C. Interestingly, antiserum raised against recombinant BgHSP‐70 protein reacted specifically not only with a 70‐kDa protein of B. gibsoni but also with a corresponding native protein of B. microti (BmHSP‐70), indicating the high degree of conservation of this protein. The BmHSP‐70 gene was then isolated and characterized and the immunoprotective properties of recombinant BgHSP‐70 (rBgHSP‐70) and rBmHSP‐70 were compared in vitro and in vivo . Both proteins had potent mitogenic effects on murine and canine mononuclear cells as evidenced by high proliferative responses and IFN‐γ production after stimulation. Immunization regimes in BALB/c and C57BL/6 mice using rBgHSP‐70 and rBmHSP‐70 elicited high antibody levels, with concurrent significant reductions in peripheral parasitaemias. Taken together, these results emphasize the potential of HSP‐70s as a molecular adjuvant vaccine.