Characterization of a rat monoclonal antibody directed against the mouse B lymphocyte IgE receptor
Rats were immunized with purified receptor for IgE (Fc/sub element of/R) obtained from an Fc/sub element of/R/sup +/ murine B cell hybridoma (01.2B2) and the rat spleen cells were fused to the rat myeloma line IR983F. One clone (B3B4) produced a monoclonal IgG/sub 2a/ antibody that specifically recognized the Fc/sub element of/R on murine B lymphocytes as determined by immunoprecipitation and SDS-PAGE analysis. FACS analysis indicated that B3B4 reacted with 01.2B2 cells or normal murine B lymphocytes but not with murine macrophages, rat spleen cells or with murine mast cell lines bearing the high affinity Fc/sub element of/R. B3B4 completely blocked murine lymphocyte rosetting of ox erythrocytes (OE) coated with mouse IgE but had not effect on rosette formation with IgG coated OE. B3B4 inhibited the binding of /sup 125/I-rat IgE and similarly IgE inhibited the binding of /sup 125/I-B3B4 to 0.1.2B2 cells in a dose dependent fashion. Saturation binding analysis indicated that B3B4 Fab' binds to twice as many sites/cell as does IgE suggesting that there are two identical determinants in the Fc/sub element of/R per IgE binding site. Like normal B lymphocytes, the number of Fc/sub element of/R on 01.2B2 cells can be upregulated by culture with rodent IgE. However, neither B3B4 nor its (Fab')/sub 2/ or Fab' fragments caused upregulation of the Fc/sub element of/R on these cells suggesting that a site specific interaction, provided only by IgE, is necessary for upregulation. These findings indicate that the B3B4 and IgE Fc binding determinants are in close proximity, but not identical; this new reagent should be very useful in additional structure, function analysis of the lymphocyte Fc/sub element of/R.
- Research Organization:
- Johns Hopkins Medical School, Baltimore, MD
- OSTI ID:
- 5326174
- Report Number(s):
- CONF-8604222-; TRN: 86-030445
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
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550201* - Biochemistry- Tracer Techniques