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In Vitro | Journal Article | Research Support, Non-U.S. Gov't

Defining antibody targets in Streptococcus oralis infection.

J P Burnie, W Brooks, M Donohoe, S Hodgetts, A al-Ghamdi, R C Matthews
J P Burnie
University Department of Medical Microbiology, Manchester Healthcare Trust, United Kingdom.
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W Brooks
University Department of Medical Microbiology, Manchester Healthcare Trust, United Kingdom.
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M Donohoe
University Department of Medical Microbiology, Manchester Healthcare Trust, United Kingdom.
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S Hodgetts
University Department of Medical Microbiology, Manchester Healthcare Trust, United Kingdom.
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A al-Ghamdi
University Department of Medical Microbiology, Manchester Healthcare Trust, United Kingdom.
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R C Matthews
University Department of Medical Microbiology, Manchester Healthcare Trust, United Kingdom.
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ABSTRACT

Immunoblotting of sera from 12 neutropenic patients with Streptococcus oralis septicemia and 18 patients with endocarditis due to viridans group streptococci revealed immunodominant S. oralis antigens at 85 and 180 kDa. The former cross-reacted with a mouse monoclonal antibody to hsp90. The latter was identified by sequencing positive clones obtained by screening a genomic expression library of S. oralis with pooled sera from patients who had been infected with S. oralis. Antibody eluted from one of these clones reacted with the 180-kDa antigen of S. oralis. Southern blotting confirmed the origin of the clone from S. oralis. The derived amino acid sequence showed 76.2% homology with the PAc protein precursor of Streptococcus mutans and 73.8% homology with the SpaA protein precursor of Streptococcus sobrinus. Epitope mapping of the derived amino acid sequence with sera from patients with viridans group streptococcal endocarditis delineated nine epitopes. Peptides 1 (TMYPNRQPGSGWDSS) and 2 (WYSLNGKIRAVDVPK), representing two of these epitopes, and peptide 3 (YEVEKPLEPAPVAPS), representing the repeat proline region, were synthesized. These three peptides were used to screen a phage antibody display library derived from a patient who had recovered from S. oralis infection. Two of the human recombinant antibodies produced (SORAL 3 and SORAL 4 against peptide 3) and a human recombinant antibody (B3.7) against the conserved epitope (LKVIRK) of hsp90 gave statistically significant protection, compared with control groups, in a mouse model of lethal S. oralis infection.

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Defining antibody targets in Streptococcus oralis infection.
J P Burnie, W Brooks, M Donohoe, S Hodgetts, A al-Ghamdi, R C Matthews
Infection and Immunity May 1996, 64 (5) 1600-1608; DOI:

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Defining antibody targets in Streptococcus oralis infection.
J P Burnie, W Brooks, M Donohoe, S Hodgetts, A al-Ghamdi, R C Matthews
Infection and Immunity May 1996, 64 (5) 1600-1608; DOI:
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