Proteolysis increases the Fc-mediated binding of murine IgG2b to human EBV-transformed B cells, but decreases the expression of Fc gamma RII and Fc epsilon RII.
We have previously described a polymorphic human Fc receptor for murine IgG2b (mIgG2b). This receptor was defined by the binding of (complexed) mIgG2b to monocytes and Epstein-Barr virus (EBV)-transformed B cells. Three per cent of normal individuals were high responders with respect to mIgG2b (mIgG2b-HR), whereas the other individuals were low responders for mIgG2b (mIgG2b-LR). In the present study we investigated the effect of proteolytic enzymes on the Fc-mediated binding of mIgG2b to EBV-B cells. Pronase, human leucocyte elastase and cathepsin G caused an increased binding (in an EA-rosetting assay) of mIgG2b to EBV-B cells from mIgG2b-HR, but not from mIgG2b-LR. Simultaneous immunofluorescence studies demonstrated that these proteolytic enzymes strongly reduced the expression of Fc gamma RII and Fc epsilon RII on these cells, whereas HLA class I or HLA class II molecules were not affected. These findings strongly suggest that binding of mIgG2b is not mediated by Fc gamma RII or Fc epsilon RII. We also studied the effect of proteolysis on mIgG2b-HR EBV-B cells from an HLA class II-negative individual. In this case EA-mIgG2b rosetting was decreased after proteolysis, suggesting that HLA class II molecules may have a role in protecting the binding site for mIgG2b against proteolytic destruction.
Department of Medicine, University Hospital Nijmegen, The Netherlands.
S
S
Holtrop
G P
GP
Rijke-Schilder
N E
NE
van Doorn
C W
CW
Jacobs
R A
RA
Koene
W J
WJ
Tax
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D056649:Cathepsin G; D002403:Cathepsins / Q000378:metabolism; D002461:Cell Line, Transformed; D015870:Gene Expression / Q000276:immunology / Q000502:physiology*; D004854:Herpesvirus 4, Human; D006801:Humans; D019272:Leukocyte Elastase; D051379:Mice; D009439:Neuraminidase / Q000378:metabolism; D010196:Pancreatic Elastase / Q000378:metabolism; D011961:Receptors, Fc / Q000276:immunology / Q000378:metabolism*; D017455:Receptors, IgE / Q000378:metabolism; D017452:Receptors, IgG / Q000378:metabolism; D012397:Rosette Formation; D012697:Serine Endopeptidases / Q000378:metabolism*
D011961:Receptors, Fc; D017455:Receptors, IgE; D017452:Receptors, IgG; D009439:Neuraminidase; D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G; C534096:Ctsg protein, mouse; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase