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A more recent version of this article appeared on March 1, 2007 Originally published In Press as doi:10.1194/jlr.C600020-JLR200 on December 11, 2006

Papers In Press, published online ahead of print December 9, 2006
J. Lipid Res., doi:10.1194/jlr.C600020-JLR200
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Submitted on November 21, 2006
Revised on December 8, 2006
Accepted on December 9, 2006

Biological effects of C-reactive protein are not due to endotoxin contamination: Evidence from toll like receptor 4 knock-down human aortic endothelial cells

Mohan R. Dasu, Sridevi Devaraj, Terry W. Du Clos, and Ishwarlal Jialal

UCDavis Med Ctr, Sacramento, CA 95817

Corresponding Author: ijialal{at}ucdavis.edu

C-reactive protein (CRP) is the prototypic marker of inflammation and a strong predictor of cardiovascular events in humans. There are questions as to the validity of the biological effects of CRP in spite of adhering to rigorous control measures minimizing endotoxin (LPS) contamination in these in vitro studies. In the present study, we addressed the key question of endotoxin contamination in CRP preparations using TLR4 knock down endothelial cells. Human aortic endothelial cells transfected with prevalidated TLR4 siRNA and scrambled siRNA controls were challenged with pleural fluid derived CRP or LPS for 12-16hrs. Secreted IL-6, IL-1b, IL-8, PAI-1 levels and eNOS activity were determined. TLR4 knock down in HAECs significantly decreased LPS induced IL-1b, IL-6, IL-8, while stimulatory effects of CRP were similar in both Sc control and TLR4 knock down cells. Furthermore, CRP significantly stimulated PAI-1 levels in both control and TLR4 transfected cells and inhibited eNOS activity, while LPS effects were negated in TLR4 transfected cells. Data presented cogently demonstrate and further confirm the biological effects of CRP on HAECs are independent of LPS and are thus due to native protein per se. This is the first study to positively implicate and authenticate the significance of earlier in vitro reports on CRP biological effects.


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