title: C-reactive protein triggers calcium signalling in human neutrophilic granulocytes via FcγRIIa in an allele-specific way.
abstract: C-reactive protein (CRP) binds to Fcγ-receptors, FcγRIIa (CD32) with high affinity and to FcγRIa (CD64) with low affinity. The binding to CD32 has been shown to be allele specific, that is, it binds to R/R131 but not to H/H131. Little is known about the cooperation of CRP and neutrophilic granulocytes (PMNs) in inflammatory reactions. The purpose of the present study was to examine CRP signalling in human PMNs, and whether this signalling is also allele specific. Cytosolic calcium of PMN was measured in a single-cell digital imaging system. Receptor expression and polymorphism were studied by real-time RT-PCR, flow cytometry and standard PCR. C-reactive protein induced cytosolic calcium signals in PMNs from homozygote R/R131 donors, but not in PMNs from heterozygote R/H131 donors. However, after the heterozygote PMNs had been incubated with IFN-γ (100 U/ml) for 2 h, both the proportion of cells responding and the size of the CRP-induced calcium signals increased. IFN-γ increased mRNA expression of CD64 about fivefold and surface protein expression of CD64 about fourfold. The calcium signal elicited by CRP was augmented by PMN adhesion to fibronectin, but almost totally abrogated by sphingosine kinase inhibitors. The signals were partly dependent on calcium influx. In conclusion, calcium signalling instigated by CRP in human PMN is FcγRIIa allele specific, as R/R131 responded to CRP, whereas R/H131 did not. However, increased expression of FcγRIa (CD64), stimulated by IFN-γ, can augment calcium signalling by CRP in low-responders. This suggests that the state of the PMNs, as well as the genetic origin, affect sensitivity for CRP.
authors.affiliation: Faculty of Health Sciences, Oslo and Akershus University College of Applied Sciences, Oslo, Norway. vigdis.aas@hioa.no
authors.forename: V
authors.initials: V
authors.lastname: Aas
authors.forename: K L
authors.initials: KL
authors.lastname: Sand
authors.forename: H-C
authors.initials: HC
authors.lastname: Asheim
authors.forename: H B
authors.initials: HB
authors.lastname: Benestad
authors.forename: J-G
authors.initials: JG
authors.lastname: Iversen
mesh_terms: D000483:Alleles; D002097:C-Reactive Protein / Q000276:immunology / Q000378:metabolism*; D020013:Calcium Signaling / Q000235:genetics* / Q000276:immunology; D005434:Flow Cytometry; D006801:Humans; D018375:Neutrophil Activation / Q000235:genetics* / Q000276:immunology; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism*; D020641:Polymorphism, Single Nucleotide; D060888:Real-Time Polymerase Chain Reaction; D017452:Receptors, IgG / Q000235:genetics* / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: C080568:Fc gamma receptor IIA; D017452:Receptors, IgG; D002097:C-Reactive Protein