Isolation and characterization of Fc gamma receptors from human placenta.
Placental extract was prepared with Tris-HCl buffer pH 7.4 containing EDTA and 2-mercaptoethanol. It agglutinated erythrocytes (E) sensitized with subagglutinating amounts of IgG antibodies from rabbits, guinea pigs and mice, but not E sensitized with IgG from chickens. E or E sensitized with F(ab)2 or IgM antibodies were not agglutinated. The agglutination of EA by the extract was inhibited by human, rabbit and guinea pig IgG, but not by bovine and porcine IgG. Aggregated IgG was more inhibitory than monomeric IgG. IgG3 was the most effective subclass. The extract inhibited the formation of EA rosettes with human mononuclear cells, but did not influence the formation of E or EAC rosettes. The significance of the disulfide bonds and the C gamma 3 and C gamma 2 regions was implied by the finding that the extract neither agglutinated E sensitized with preparations of mildly reduced and alkylated IgG, nor with Facb fragments. These preparations did not inhibit the agglutination. The results strongly indicated that the active component was solubilized placental Fc receptor (FcR). Functionally active FcR was purified by affinity chromatography using aggregated human IgG coupled to Sepharose 4B. SDS-PAGE if the purified FcR under reducing conditions showed one distance band corresponding to approx. 47,000 daltons. The band neither consisted o Ig fragments nor Clq. A rabbit antiserum against the FcR inhibited the agglutination of EA by the extract and stained th FcR-positive areas in placenta.
D002846:Chromatography, Affinity; D005260:Female; D006384:Hemagglutination; D006801:Humans; D007074:Immunoglobulin G; D010920:Placenta / Q000276:immunology*; D011247:Pregnancy; D011961:Receptors, Fc / Q000302:isolation & purification* / Q000502:physiology; D012397:Rosette Formation
D007074:Immunoglobulin G; D011961:Receptors, Fc