Detection of lymphocyte Fc gamma receptor-blocking factors by the EA rosette inhibition assay. Refinement of the conventional method and development of a novel flow-cytometric assay.
Serum factors which interact with human peripheral blood lymphocyte Fc gamma receptors (Fc gamma Rs) may be detected in vitro by the EA rosette inhibition assay (EARIA). This assay has been used to detect circulating immune complexes and certain alloantibodies directed against cell surface antigens situated in close proximity to Fc gamma Rs. Three main types of FcR-blocking factor have been demonstrated by the EARIA in human serum following exposure to alloantigens. A strong correlation was observed between the presence of one of these FcR-blocking factors (FcBF1) and human renal allograft survival. This factor was previously shown to bind preferentially to CD32+ B cells and to inhibit antibody synthesis. In this study we have shown that detection of FcBF1 by the EARIA depends on the type of erythrocyte and on the amount of antibody used to sensitise the erythrocytes. Furthermore, we have developed a flow-cytometric version of the EARIA which is rapid, reproducible and, most importantly, objective. Inter-laboratory comparisons using this standardised EARIA should now be possible.
Sandilands
G P
GP
University Department of Pathology, Western Infirmary, Glasgow, Scotland, UK.
Greer
M R
MR
Chisholm
S E
SE
McKay
I C
IC
Downie
I
I
McMillan
M A
MA
MacSween
R N
RN
D004912:Erythrocytes / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D016030:Kidney Transplantation / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D017452:Receptors, IgG / Q000037:antagonists & inhibitors*; D012397:Rosette Formation / Q000379:methods*
D017452:Receptors, IgG