Characterization of a rat monoclonal antibody directed against the mouse B lymphocyte IgE receptor

Conference · Sat Mar 01 04:00:00 EST 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5326174

Rao, M; Lee, W T; Conrad, D H

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-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:3
• Country of Publication: United States
• Language: English

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550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AFFINITY
ANIMAL CELLS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CELL CULTURES
CHEMICAL COMPOSITION
CLONE CELLS
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DOSE-RESPONSE RELATIONSHIPS
ELECTRON CAPTURE RADIOISOTOPES
ERYTHROCYTES
GLOBULINS
IMMUNOGLOBULINS
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LEUKOCYTES
LYMPHOCYTES
MACROPHAGES
MAMMALS
MAST CELLS
MATERIALS
MEMBRANE PROTEINS
MICE
MONOCLONAL ANTIBODIES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PHAGOCYTES
PROTEINS
RADIOISOTOPES
RATS
REACTION KINETICS
RECEPTORS
RODENTS
SOMATIC CELLS
SPLEEN CELLS
TRACER TECHNIQUES
VERTEBRATES