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Interaction of monoclonal antibodies with the IgE-receptor on rat mast cells and rat basophilic leukemia (RBL) cells

  • Histamine and Kinins
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Abstract

The close relation between rat mast cells and rat basophilic leukemia (RBL) cells with regard to the presence of receptors for IgE and Fcγ led us to generate monoclonal antobodies directed against cell surface antigens. Hybridomas were obtained by the fusion of NS1 mouse myeloma cells with murine spleen and lymph node cells. The culture supernatants were assayed by two ELISA techniques: a) for the production of mouse immunoglobulin in general and b) for antibodies directed against surface antigens of RBL cells. For this purpose RBL cells were attached to polyvinyl chloride microtitre plates. Eight hybrids produced antibodies directed against surface antigens on RBL cells. Hybrids were cloned and characterized with regard to their isotype and light chains. All eight clones secreted IgM with K light chains. Immunofluorescence studies performed with RBL cells revealed that all eight antibodies were able to show a specific fluorescence. Furthermore, four of these eight antibodies also showed a specific fluorescence with purified rat mast cells. These four antibodies were analyzed as to their ability of interacting with the IgE-receptor on RBL cells and purified rat mast cells. They reduced the binding rate of radiolabelled rat IgE to RBL and rat mast cells. A mutual inhibition of the passive cutaneous anaphylaxis (PCA) reaction in the rat by either mixing mouse reaginic serum directed against 2,4-dinitrophenol bovine serum albumin (DNP-BSA) or by mixing monoclonal mouse anti-DNP IgE with the monoclonal mouse anti-cell surface (rat basophilic leukemia, rat mast cell) IgM was determined. The histamine release of rat mast cells obtained from Nippo-strongylus brasiliensis infected rats was markedly reduced in the presence of two monoclonal anti cell surface antibodies. Our data suggest that monoclonal antibodies with specificity for the IgE receptor (inhibition of IgE-binding) can be distinguished from those which bind closely to the IgE-receptor but also modulate the histamine release from presensitized mast cells.

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Abbreviations

DMEM:

Dulbecco's modified Eagle's Medium

DMSO:

dimethylsulfoxide

DPBS:

Dulbecco's phosphate buffered saline

FCS:

fetal calf serum

HAT:

hypoxanthine — aminopterine — thymidine

HT:

hypoxanthine — thymidine

ION:

calcium ionophore A 23187

MAC:

monoclonal antibody against cell surface antigens

MRS:

mouse reaginic serum

Nb:

Nippostrongylus brasiliensis

PCA:

passive cutaneous anaphylaxis

PEG:

polyethyleneglycoll

RAM:

rabbit anti mouse

RBL cells:

rat basophilic leukemia cells

RMC:

rat mast cells

TRIS:

tris-(hydroxymethyl)aminomethane

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Authors and Affiliations

  1. Lehrstuhl Med. Mikrobiologie und Immunologie—AG Infektabwehrmechanismen, RUHR-Universität Bochum, Universitätsstr. 150, Postfach 10 21 48, 4630, Bochum 1, FRG

    G. Micklefield,  W. König, Ph. Pfeiffer & A. Bohn

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  1. G. Micklefield
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  2. W. König
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  3. Ph. Pfeiffer
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  4. A. Bohn
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in partial fulfilment of G.M. M.D.-thesis supported by DFG Kö 427-7/2

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Micklefield, G., König, W., Pfeiffer, P. et al. Interaction of monoclonal antibodies with the IgE-receptor on rat mast cells and rat basophilic leukemia (RBL) cells. Agents and Actions 17, 418–426 (1986). https://doi.org/10.1007/BF01965508

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  • DOI: https://doi.org/10.1007/BF01965508

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