An intact phosphocholine binding site is necessary for transgenic rabbit C-reactive protein to protect mice against challenge with platelet-activating factor.
C-reactive protein (CRP), an acute phase protein in humans and rabbits, is part of the innate immune system. The role of CRP in host defense has been thought to be largely due to its ability to bind phosphocholine, activate complement, and interact with IgGRs (FcgammaRs). We have shown previously that transgenic rabbit CRP (rbCRP) protects mice from lethal challenges with platelet-activating factor (PAF). To investigate the mechanism of this protection, we created additional lines of transgenic mice that express either wild-type rbCRP, a variant of rbCRP with altered complement activation activity (Y175A), or a variant of rbCRP unable to bind phosphocholine (F66Y/E81K). In the current study, these lines were challenged with a single injection of PAF and their survival monitored. Mice expressing wild-type and Y175A rbCRP were protected against challenge by PAF whereas mice expressing F66Y/E81K rbCRP were not. Treatment with cobra venom factor did not affect survival, confirming the results with the Y175A rbCRP variant and indicating that complement activation was not required to mediate protection. Both wild-type rbCRP and Y175A rbCRP were capable of binding PAF in vitro whereas F66Y/E81K rbCRP was not. Although other interpretations are possible, our results suggest that the protective effect of rbCRP against PAF is due to sequestration of PAF.
Department of Biochemistry, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA.
Steven
S
Black
Augusta
A
Wilson
David
D
Samols
D000818:Animals; D001665:Binding Sites / Q000235:genetics; D002097:C-Reactive Protein / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D003167:Complement Activation / Q000502:physiology; D004040:Dietary Carbohydrates / Q000008:administration & dosage; D004044:Dietary Proteins / Q000008:administration & dosage; D018698:Glutamic Acid / Q000235:genetics; D008239:Lysine / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010649:Phenylalanine / Q000235:genetics; D010767:Phosphorylcholine / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000633:toxicity*; D011485:Protein Binding / Q000235:genetics; D011817:Rabbits; D016019:Survival Analysis; D014443:Tyrosine / Q000235:genetics
D004040:Dietary Carbohydrates; D004044:Dietary Proteins; D010972:Platelet Activating Factor; D010767:Phosphorylcholine; D018698:Glutamic Acid; D014443:Tyrosine; D010649:Phenylalanine; D002097:C-Reactive Protein; D008239:Lysine