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Antibodies against C-reactive protein cross-react with 60-kilodalton heat shock proteins.
Udvarnoki, Katalin Cervenak, László Uray, Katalin Hudecz, Ferenc Kacskovics, Imre Spallek, Ralf Singh, Mahavir Füst, George Prohászka, Zoltán
Udvarnoki, Katalin
Cervenak, László
Uray, Katalin
Hudecz, Ferenc
Kacskovics, Imre
Spallek, Ralf
Singh, Mahavir
Füst, George
Prohászka, Zoltán
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2007-04
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Abstract
C-reactive protein (CRP) is an acute-phase reactant frequently used in histochemistry as a marker of ongoing inflammation. Furthermore, CRP is a powerful biomarker for the prediction of coronary artery disease risk. Heat-shock protein 60 (Hsp60) and CRP are complement-activating molecules, and the effect of their interactions on the regulation of complement activation was studied. However, during the first experiments, we learned that polyclonal anti-CRP antibodies cross-react with Hsp60. Therefore, the aim of our present study was to analyze the cross-reactivity of anti-CRP antibodies (Ab) with Hsp60 in solid-phase enzyme immune assays, in epitope studies using a series of overlapping synthetic peptides, and in Ouchterlony analyses. We found that three different commercial rabbit polyclonal antibodies and two monoclonal (9C9 and CRP-8) anti-CRP antibodies specifically recognize recombinant human Hsp60 and recombinant Mycobacterium tuberculosis Hsp65, respectively. Hsp60 was found to inhibit the binding of anti-CRP polyclonal Ab to Hsp60. Six epitope regions of Hsp60 were recognized by the anti-CRP antibodies, and one region (amino acids [AA] 218 to 232) was recognized by monoclonal antibodies CRP-8 and 9C9. This epitope region of Hsp60 displays 26.6% amino acid identity to CRP AA region 77 to 90. These data suggest that the B-cell epitopes shared between CRP and Hsp60 give rise to a true mimicry-based cross-reaction and the induction of cross-reactive antibodies. Our study underlines the importance of thorough study design and careful interpretation of results while using polyclonal anti-CRP antibodies for histochemistry, especially at low dilutions. Furthermore, analytical interference with Hsp60 in CRP assays should also be tested.
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Antibodies against C-reactive protein cross-react with 60-kilodalton heat shock proteins. 2007, 14 (4):335-41 Clin. Vaccine Immunol.
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en
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1556-6811