Abstract
C-reactive protein (CRP) is an established marker of inflammation and has been proposed to play a proinflammatory role in pathologies of several diseases. CRP is primarily produced by the liver and released into circulation as a pentameric molecule composed of five identical subunits. It has been suggested that the activation of the proinflammatory actions of CRP requires sequential conformational changes triggered by local inflammatory conditions. These include the dissociation into the subunit form (monomeric CRP, mCRP) and further reduction of the intra-subunit disulfide bond of mCRP. This model predicts that mCRP is the primary isoform present in inflamed but not healthy tissues, however the supporting evidence is lacking. Herein, we stained tissue samples across multiple anatomical locations from several types of human diseases with highly selective monoclonal antibodies that can differentiate CRP and mCRP. The results indicated that mCRP is the predominant form existing in the lesions. Further immunoblotting of the patient tissue samples revealed the potential presence of reduced mCRP. Together, we conclude that mCRP but not CRP is the major isoform present in local inflammatory lesions, supporting the so-called cascading model of CRP function and regulation.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Allin, K. H., Nordestgaard, B. G. (2011) Elevated C-reactive protein in the diagnosis, prognosis, and cause of cancer. Crit. Rev. Clin. Lab. Sci. 48, 155–170.
Ciubotaru, I., Potempa, L. A., Wander, R. C. (2005) Production of modified C-reactive protein in U937-derived macrophages. Exp. Biol. Med. (Maywood) 230, 762–770.
Eisenhardt, S. U., Habersberger, J., Murphy, A., Chen, Y. C., Woollard, K. J., Bassler, N., Qian, H., von Zur Muhlen, C., Hagemeyer, C. E., Ahrens, I., Chin-Dusting, J., Bobik, A., Peter, K. (2009) Dissociation of pentameric to monomeric C-reactive protein on activated platelets localizes inflammation to atherosclerotic plaques. Circ. Res. 105, 128–137.
Eisenhardt, S. U., Thiele, J. R., Bannasch, H., Stark, G. B., Peter, K. (2009) C-reactive protein: how conformational changes influence inflammatory properties. Cell Cycle 8, 3885–3892.
Filep, J. G. (2009) Platelets affect the structure and function of C-reactive protein. Circ. Res. 105, 109–111.
Jabs, W. J., Logering, B. A., Gerke, P., Kreft, B., Wolber, E. M., Klinger, M. H., Fricke, L., Steinhoff, J. (2003) The kidney as a second site of human C-reactive protein formation in vivo. Eur. J. Immunol. 33, 152–161.
Ji, S. R., Ma, L., Bai, C. J., Shi, J. M., Li, H. Y., Potempa, L. A., Filep, J. G., Zhao, J., Wu, Y. (2009) Monomeric C-reactive protein activates endothelial cells via interaction with lipid raft microdomains. FASEB J. 23, 1806–1816.
Ji, S. R., Wu, Y., Potempa, L. A., Liang, Y. H., Zhao, J. (2006) Effect of modified C-reactive protein on complement activation: a possible complement regulatory role of modified or monomeric C-reactive protein in atherosclerotic lesions. Arterioscler. Thromb. Vasc. Biol. 26, 935–941.
Ji, S. R., Wu, Y., Potempa, L. A., Qiu, Q., Zhao, J. (2006) Interactions of C-reactive protein with lowdensity lipoproteins: implications for an active role of modified C-reactive protein in atherosclerosis. Int. J. Biochem. Cell Biol. 38, 648–661.
Ji, S. R., Wu, Y., Zhu, L., Potempa, L. A., Sheng, F. L., Lu, W., Zhao, J. (2007) Cell membranes and liposomes dissociate C-reactive protein (CRP) to form a new, biologically active structural intermediate: mCRP(m). FASEB J. 21, 284–294.
Khreiss, T., Jozsef, L., Potempa, L. A., Filep, J. G. (2004) Conformational rearrangement in C-reactive protein is required for proinflammatory actions on human endothelial cells. Circulation 109, 2016–2022.
Khreiss, T., Jozsef, L., Potempa, L. A., Filep, J. G. (2005) Loss of pentameric symmetry in C-reactive protein induces interleukin-8 secretion through peroxynitrite signaling in human neutrophils. Circ. Res. 97, 690–697.
Khreiss, T., Jozsef, L., Potempa, L. A., Filep, J. G. (2004) Opposing effects of C-reactive protein isoforms on shear-induced neutrophil-platelet adhesion and neutrophil aggregation in whole blood. Circulation 110, 2713–2720.
Lauer, N., Mihlan, M., Hartmann, A., Schlotzer-Schrehardt, U., Keilhauer, C., Scholl, H. P., Charbel Issa, P., Holz, F., Weber, B. H., Skerka, C., Zipfel, P. F. (2011) Complement regulation at necrotic cell lesions is impaired by the age-related macular degeneration-associated factor-H His402 risk variant. J. Immunol. 187, 4374–4383.
Ma, X., Ji, S. R., Wu, Y. (2013) Regulated conformation changes in C-reactive protein orchestrate its role in atherogenesis. Chinese Sci. Bull. 58, 1642–1649.
Mihlan, M., Blom, A. M., Kupreishvili, K., Lauer, N., Stelzner, K., Bergstrom, F., Niessen, H. W., Zipfel, P. F. (2011) Monomeric C-reactive protein modulates classic complement activation on necrotic cells. FASEB J. 25, 4198–4210.
Molins, B., Pena, E., de la Torre, R., Badimon, L. (2011) Monomeric C-reactive protein is prothrombotic and dissociates from circulating pentameric C-reactive protein on adhered activated platelets under flow. Cardiovasc. Res. 92, 328–337.
Pepys, M. B., Hirschfield, G. M. (2003) C-reactive protein: a critical update. J. Clin. Invest. 111, 1805–1812.
Singh, S. K., Suresh, M. V., Voleti, B., Agrawal, A. (2008) The connection between C-reactive protein and atherosclerosis. Ann. Med. 40, 110–120.
Sjowall, C., Bengtsson, A. A., Sturfelt, G., Skogh, T. (2004) Serum levels of autoantibodies against monomeric C-reactive protein are correlated with disease activity in systemic lupus erythematosus. Arthritis. Res. Ther. 6, R87–94.
Sjowall, C., Cardell, K., Bostrom, E. A., Bokarewa, M. I., Enocsson, H., Ekstedt, M., Lindvall, L., Fryden, A., Almer, S. (2012) High prevalence of autoantibodies to C-reactive protein in patients with chronic hepatitis C infection: association with liver fibrosis and portal inflammation. Hum. Immunol. 73, 382–388.
Wang, M. S., Messersmith, R. E., Reed, S. M. (2012) Membrane curvature recognition by C-reactive protein using lipoprotein mimics. Soft Matter. 8, 3909–3918.
Wang, M. Y., Ji, S. R., Bai, C. J., El Kebir, D., Li, H. Y., Shi, J. M., Zhu, W., Costantino, S., Zhou, H. H., Potempa, L. A., Zhao, J., Filep, J. G., Wu, Y. (2011) A redox switch in C-reactive protein modulates activation of endothelial cells. FASEB J. 25, 3186–3196.
Wang, M. Y., Zhou, H. H., Zhang, S. C., Hui, F., Zhu, W., Su, H. X., Guo, H. Y., Li, X. W., Ji, S. R., Wu, Y. (2014) Recurrent mutations at C-reactive protein gene promoter SNP position -286 in human cancers. Cell Res., 24, 505–508.
Wettero, J., Nilsson, L., Jonasson, L., Sjowall, C. (2009) Reduced serum levels of autoantibodies against monomeric C-reactive protein (CRP) in patients with acute coronary syndrome. Clin. Chim. Acta 400, 128–131.
Wu, Y., Ji, S. R., Wang, H. W., Sui, S. F. (2002) Study of the spontaneous dissociation of rabbit C-reactive protein. Biochemistry (Mosc) 67, 1377–1382.
Wu, Y., Wang, H. W., Ji, S. R., Sui, S. F. (2003) Two-dimensional crystallization of rabbit C-reactive protein monomeric subunits. Acta Crystallogr. D. Biol. Crystallogr. 59, 922–926.
Yasojima, K., Schwab, C., McGeer, E. G., McGeer, P. L. (2001) Generation of C-reactive protein and complement components in atherosclerotic plaques. Am. J. Pathol. 158, 1039–1051.
Ying, S. C., Gewurz, H., Kinoshita, C. M., Potempa, L. A., Siegel, J. N. (1989) Identification and partial characterization of multiple native and neoantigenic epitopes of human C-reactive protein by using monoclonal antibodies. J. Immunol. 143, 221–228.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
About this article
Cite this article
Shi, P., Li, XX., Zhu, W. et al. Immunohistochemical Staining Reveals C-Reactive Protein Existing Predominantly as Altered Conformation Forms in Inflammatory Lesions. BIOLOGIA FUTURA 65, 265–273 (2014). https://doi.org/10.1556/ABiol.65.2014.3.3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1556/ABiol.65.2014.3.3