Antibodies against C-reactive protein cross-react with 60-kilodalton heat shock proteins.
dc.contributor.author | Udvarnoki, Katalin | |
dc.contributor.author | Cervenak, László | |
dc.contributor.author | Uray, Katalin | |
dc.contributor.author | Hudecz, Ferenc | |
dc.contributor.author | Kacskovics, Imre | |
dc.contributor.author | Spallek, Ralf | |
dc.contributor.author | Singh, Mahavir | |
dc.contributor.author | Füst, George | |
dc.contributor.author | Prohászka, Zoltán | |
dc.date.accessioned | 2008-02-21T14:09:36Z | |
dc.date.available | 2008-02-21T14:09:36Z | |
dc.date.issued | 2007-04 | |
dc.identifier.citation | Antibodies against C-reactive protein cross-react with 60-kilodalton heat shock proteins. 2007, 14 (4):335-41 Clin. Vaccine Immunol. | en |
dc.identifier.issn | 1556-6811 | |
dc.identifier.pmid | 17301219 | |
dc.identifier.doi | 10.1128/CVI.00155-06 | |
dc.identifier.uri | http://hdl.handle.net/10033/18872 | |
dc.description.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. | |
dc.language.iso | en | en |
dc.subject.mesh | Animals | en |
dc.subject.mesh | Antibodies | en |
dc.subject.mesh | Bacterial Proteins | en |
dc.subject.mesh | Binding Sites, Antibody | en |
dc.subject.mesh | C-Reactive Protein | en |
dc.subject.mesh | Chaperonin 60 | en |
dc.subject.mesh | Chaperonins | en |
dc.subject.mesh | Cross Reactions | en |
dc.subject.mesh | Humans | en |
dc.subject.mesh | Mice | en |
dc.subject.mesh | Mycobacterium tuberculosis | en |
dc.subject.mesh | Rabbits | en |
dc.title | Antibodies against C-reactive protein cross-react with 60-kilodalton heat shock proteins. | en |
dc.type | Article | en |
dc.contributor.department | Third Department of Medicine, Semmelweis University, H-1125 Budapest, Kútvölgyi st. 4, Hungary. | en |
dc.identifier.journal | Clinical and vaccine immunology : CVI | en |
refterms.dateFOA | 2018-06-13T03:46:06Z | |
html.description.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. |