Devaraj, and S. Kuta and L. Willerson, and E. Calabro, D. Chang, J. Zhang, M. Sun, D. Samols, and I. Agrawal, D. Enocsson, C. Skogh, M. Eloranta, L. Voleti and A. Migliorini and H. Theofilopoulos, R. Baccala, B. Beutler, and D. View at: Google Scholar W. Hutchinson, G. Noble, P. Hawkins, and M. Pepys and M. Singh, M. Suresh, B. Voleti, and A. Allin and B. Yousuf, B. Mohanty, S. Martin et al. Brand, S. Page, G. Rogler et al. Thurberg and T. Marumo, V. Schini-Kerth, B. Fisslthaler, and R.
Pasceri, J. Taylor, and D. Swafford Jr. Bratz, J. Knudson et al. H—H, Pepys, P. Hawkins, M. Kahan et al. Eisenhardt, M. Schwarz, N. Bassler, and K. Schallner et al. Habersberger, A. Murphy et al. Khreiss, L. Potempa, and J.
Zouki, M. Beauchamp, C. Baron, and J. Zouki, B. Haas, J. Chan, L. Potempa, B. Maldonado, P. Laurent, E. Zemel, and H. Kresl, L. Potempa, and B. Potempa, Q. Qiu, and J. Pepys, J. Gallimore, J. Lloyd et al. Lane, N. Wassef, S. Poole et al. Habersberger, D. Braig et al. Habersberger, F. Strang, A. Scheichl et al. Scheichl, Y.
Chen et al. Eisenhardt, Y. Schmidt, G. Karaxha et al. Teslenko, M. Rogers, and J. Edsfeldt, N. Ko et al. Hossain, J. Motie, K. Schaul, and L. View at: Google Scholar H. Li, J. Wang, Y. Wu et al. Bray, N. Samberg, H. Gewurz, L. Potempa, and A. View at: Google Scholar N. Samberg, R. Bray, H. Gewurz, A. Landay, and L. Rees, H. Gewurz, J. Siegel, J.
Coon, and L. Murphy, L. Baum, and K. Dong and J. View at: Google Scholar J. Gould and J. Diehl, G. Haines III, J. Radosevich, and L. Owens III and N. Bang, L. Marnell, C. Mold et al. Lu, L. Marnell, K. Marjon, C. Mold, T. Du Clos, and P. Ji, L. Ma, C. Bai et al. Colotta, F. Re, N. Polentarutti, S. Sozzani, and A. C-reactive protein is produced by a small number of normal human peripheral blood lymphocytes. J Exp Med —6. Release of C-reactive protein in response to inflammatory cytokines by human adipocytes: linking obesity to vascular inflammation.
J Am Coll Cardiol —3. Brain Res —9. The kidney as a second site of human C-reactive protein formation in vivo. Eur J Immunol — Expression of C-reactive protein in human lung epithelial cells and upregulation by cytokines and carbon particles. Inhal Toxicol — Acute phase proteins with special reference to C-reactive protein and related proteins pentaxins and serum amyloid A protein.
Adv Immunol — C-reactive protein levels in patients with rheumatoid arthritis: the impact of therapy. Clin Rheumatol —4. Povoa P. C-reactive protein: a valuable marker of sepsis. Intensive Care Med — C-reactive protein kinetics after major surgery.
Anesth Analg —9. C-reactive protein and lesion morphology in patients with acute myocardial infarction. Circulation —5. STAT3 participates in transcriptional activation of the C-reactive protein gene by interleukin J Biol Chem —9. Interferon-alpha mediates suppression of C-reactive protein: explanation for muted C-reactive protein response in lupus flares? Arthritis Rheum — JAMA — Voleti B, Agrawal A.
Statins and nitric oxide reduce C-reactive protein production while inflammatory conditions persist. Mol Immunol —6. Immunology — Structural recognition and functional activation of FcgammaR by innate pentraxins. Nature — Monomeric C-reactive protein activates endothelial cells via interaction with lipid raft microdomains.
Loss of pentameric symmetry of C-reactive protein is associated with promotion of neutrophil-endothelial cell adhesion. Loss of pentameric symmetry in C-reactive protein induces interleukin-8 secretion through peroxynitrite signaling in human neutrophils. Circ Res —7. Conformational rearrangement in C-reactive protein is required for proinflammatory actions on human endothelial cells.
Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell — Fibrinogen deposition at the postischemic vessel wall promotes platelet adhesion during ischemia-reperfusion in vivo. Blood — PubMed Abstract Google Scholar. Topological localization of monomeric C-reactive protein determines proinflammatory endothelial cell responses.
Platelets and the immune continuum. Nat Rev Immunol — Monomeric C-reactive protein is prothrombotic and dissociates from circulating pentameric C-reactive protein on adhered activated platelets under flow. J Thromb Haemost — 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 — Kushner I, Kaplan MH. Studies of acute phase protein. An immunohistochemical method for the localization of Cx-reactive protein in rabbits. Association with necrosis in local inflammatory lesions. C-reactive protein binds to apoptotic cells, protects the cells from assembly of the terminal complement components, and sustains an antiinflammatory innate immune response: implications for systemic autoimmunity.
Monomeric CRP contributes to complement control in fluid phase and on cellular surfaces and increases phagocytosis by recruiting factor H. Cell Death Differ — Monomeric C-reactive protein inhibits renal cell-directed complement activation mediated by properdin. Monomeric C-reactive protein modulates classic complement activation on necrotic cells. Eltzschig HK, Eckle T. Ischemia and reperfusion — from mechanism to translation. Nat Med — Myocardial ischemia-reperfusion injury: a neglected therapeutic target.
J Clin Invest — C-reactive protein and complement are important mediators of tissue damage in acute myocardial infarction. C-reactive protein colocalizes with complement in human hearts during acute myocardial infarction. C-reactive protein exacerbates renal ischemia-reperfusion injury. Acute inflammation is persistent locally in burn wounds: a pivotal role for complement and C-reactive protein. J Burn Care Res — A conformational change of C-reactive protein in burn wounds unmasks its proinflammatory properties.
Int Immunol — Am J Transplant 18 Suppl 1 — Cecka JM. Early rejection: determining the fate of renal transplants. Transplant Proc —4. The incidence and impact of early rejection episodes on graft outcome in recipients of first cadaver kidney transplants. Transplantation —8.
Transplantation —9. Non-self recognition by monocytes initiates allograft rejection. Transplant Proc — Cold ischemia and the reduced long-term survival of cadaveric renal allografts.
Kidney Int —8. The registry of the International Society for Heart and Lung Transplantation: thirteenth official pediatric lung and heart-lung transplantation report — J Heart Lung Transplant — Contribution of prolonged ischemia and donor age to chronic renal allograft dysfunction.
J Am Soc Nephrol — Opelz G, Wujciak T. The influence of HLA compatibility on graft survival after heart transplantation. The Collaborative Transplant Study. N Engl J Med —9. Antibody-mediated rejection across solid organ transplants: manifestations, mechanisms, and therapies. Toll-like receptors: critical proteins linking innate and acquired immunity. Nat Immunol — Pathogen recognition and innate immunity.
Ross R. Atherosclerosis — an inflammatory disease. Libby P, Ridker PM. Inflammation and atherosclerosis: role of C-reactive protein in risk assessment. More detailed protocol information is provided in Supplementary material online.
The MP gate was defined by size, using 0. Samples were run under native PAGE conditions and blotted onto nitrocellulose membranes as previously described. Whole blood was spun at g for 15 min. The top two-thirds of plasma were collected and remaining cells pelleted by a 2 min spin at 13 g. Concentrated MPs were then prepared by centrifugation at 30 g for 90 min. After centrifugation, an MP-depleted plasma sample was collected from the top of the preparation and the remainder discarded.
Samples were run on a native PAGE and blotted as described earlier. The membrane was stained with the specific mCRP antibody clone 9C9. The study population was divided into four clinical groups. Patient groups were defined on the basis of clinical presentation and angiographic findings. The third patient group included patients who were found to have non-obstructive coronary artery disease and did not undergo PCI.
The final group were patients with no angiographic evidence of coronary artery disease. All patients were enrolled 24—48 h after angiography. The previously documented rapid time course of pCRP dissociation 11 , 12 ensured that there was adequate time for this process to occur.
The clinical inclusion criteria were age 18—80 years, coronary artery disease status established by angiography, and willing and able to provide informed consent.
Following enrolment, patients were allocated to the appropriate disease group. All patients gave informed consent before enrolment. Western blot samples were run individually or in pairs, therefore results were not quantified and were recorded as either positive or negative for the presence of mCRP.
Microscope settings were obtained with controls in which primary antibody had been omitted. All statistical analyses were performed using GraphPad Prism v5.
All experiments were performed at least three times. The MP-depleted sample was unable to produce a significant amount of mCRP, consistent with the substantially reduced MP count as measured by flow cytometry. Concentrated MPs MP conc.
Flow cytometry was performed to quantitate the concentration of MPs in each sample concentrated and depleted. Membranes were probed for mCRP using antibody clone 9C9. All results were obtained at least three times with representative western blots shown above. The lipidomic profile of MPs derived from activated platelets were analysed by mass spectrometry to investigate for the presence of lysophosphatidylcholine LPC , which has previously been shown to be essential for pCRP binding 17 and conversion to mCRP.
This process did not occur in the MP-depleted supernatant control. For this study, four groups of patients across the spectrum of coronary artery disease were recruited. The final groups consisted of patients with stable coronary artery disease not requiring PCI and patients with angiographically normal coronary arteries. Patient characteristics were comparable across the groups, as shown in Table 1. There were no other significant differences between the groups. Collected samples were analysed by western blotting and flow cytometry for the presence or absence of mCRP.
A representative blot is shown in Figure 3 A. MP analysis by flow cytometry was also performed on each of the patient groups. These data are shown in Figure 4 with a representative flow cytometry histogram. These results were not significantly affected if normalized for circulating MP numbers data not shown. A MPs were analysed by flow cytometry. B Representative flow cytometry histograms from each patient group with secondary control shown.
We next investigated whether MPs could transfer mCRP to intact cells, potentially acting as mediators for the transmission of a pro-inflammatory signal. This clearly shows that MPs bind mCRP and are able to subsequently transfer it to the activated endothelial cells. MPs transfer mCRP to endothelial cells. In this study, we demonstrate that the dissociation process of pCRP to monomeric subunits occurs following its binding to MPs.
This finding is consistent with the previous discovery that this process takes place on activated cell membranes, either in model systems or in vitro. This finding is supported by a recent study that also identified CRP on circulating MPs, 19 however in this study the conformational status of CRP was not assessed. This is the first study to our knowledge in which mCRP specifically has been detected in the peripheral circulation. This is a critical advance in establishing the biological relevance of this isoform and may resolve some of the conflicts currently surrounding the pathological role of CRP in general.
Although circulating p CRP levels have been shown to correlate with individuals' cardiovascular risk, 3 population studies investigating a direct causal role have generally been inconclusive.
This is despite the fact that CRP has been detected in atherosclerotic plaques, 11 infarcted myocardium, 18 and has pro-inflammatory effects on the vascular endothelium.
The ability of pCRP to dissociate to monomers with unique biological properties was recognized previously, 10 but the relevance of this process to biological systems has not been fully explored. Upon agonist stimulation, platelets undergo marked changes in membrane shape and phospholipid composition.
Phospholipase A2 PLA2 activation mostly cytosolic PLA2 is essential to enable the production of potent pro-thrombotic eicosanoid mediators such as thromboxane A2, which promote localized platelet recruitment. It is these rapid changes in the membrane structure of platelets following activation that lead them to shed a large number of MPs. Indeed, the majority of circulating MPs have been found to derive from platelets.
The demonstration of a plausible mechanism of in vivo formation of mCRP 11 coupled with the discovery that mCRP can be detected on MPs potentially marks a paradigm shift in our understanding of CRP in inflammation.
These include complement component C1q fixation 12 , 18 , 26 and activation of monocytes 11 , 27 and platelets, 28 , 29 all of which contribute to myocardial ischaemia and reperfusion injury. The pathological role of circulating MPs has not been well established, although they have been implicated in the pathogenesis of inflammatory arthritis 30 and atherosclerosis.
MPs carrying proteins such as mCRP are able to engage cell surfaces through phagocytosis, ligand engagement, or membrane fusion. Recent work has shown that MPs play an important transport role for microRNAs 33 and it is plausible that a similar role is played for insoluble proteins such as mCRP.
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