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Protective Immunity against Q Fever Induced with a Recombinant P1 Antigen Fused with HspB of Coxiella burnetii
Author(s) -
LI QINGFENG,
NIU DONGSHENG,
WEN BOHAI,
CHEN MEILING,
QIU LING,
ZHANG JINGBO
Publication year - 2005
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1355.021
Subject(s) - q fever , coxiella burnetii , immunity , antigen , recombinant dna , virology , immunology , biology , immune system , genetics , gene
A bstract : The gene fragments encoding outer membrane protein 1 (P1) and heat‐shock protein B (HspB) amplified from genomic DNA of Coxiella burnetii Xinqiao by PCR were inserted into prokaryotic expression vector pQE30 to construct recombinant expression plasmids pQE30/ p1 and pQE30/ hspB , respectively. The p1 fragment from pQE30/ p1 was ligated with hspB of pQE30/ hspB to construct pQE30/ p1‐hspB . Recombinant proteins, P1, HspB, and P1‐HspB, were expressed in Escherichia coli cells transformed with pQE30/ p1 , pQE30/ hspB , and pQE30/ p1‐hspB , respectively. The purified recombinant proteins and whole‐cell antigen (WCA) of C. burnetii were used to immunize BALB/c mice. The antibody detection, T‐cell proliferation assay, and cytokine detection demonstrated that the animals immunized with P1‐HspB or WCA exhibited stronger humoral and cellular immune responses compared with animals immunized with P1 or HspB individually. Seven days after challenge of 10‐fold 50% infection dose of C. burnetii , mice were euthanized and their spleens were collected. The splenic weights of mice immunized with P1‐HspB or WCA were significantly lighter than that of mice immunized with P1 or HspB. By real‐time PCR assay, the coxiella loads of spleens of mice immunized with P1‐HspB or WCA were also significantly lower than that of mice immunized with P1 or HspB. The data from this study indicate that fusion antigen P1‐HspB is a good immunogen for eliciting immunoresponses against C. burnetii , and it may be a more suitable candidate for preparing subunit vaccine against Q fever.