Monoclonal Antibodies against Vero Cells That Protect against Diphtheria Toxin

Mice were immunized with a cell line (Vero) that possesses a high number of membrane receptors for diphtheria toxin. Spleen cells from these mice were fused with SP2/0-Ag14 cells and two cell lines (1A2 and 2D2) isolated by screening for the ability of their secreted antibodies to inhibit binding of...

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Bibliographic Details
Main Authors: Roennberg, Bengt J, Lidgerding, Burt C, Middlebrook, John L
Format: Report
Language:English
Subjects:
ANTIBODIES
BACTERIAL TOXINS
Biochemistry
Biology
BIOSYNTHESIS
CELLS
CELLS(BIOLOGY)
DORYNEBACTERIUM DIPHTHERIAE
ENZYMES
INHIBITION
ISOLATION
LABELED SUBSTANCES
MICE
MONOCLONAL ANTIBODIES
PHOSPHOLIPIDS
PROTEINS
RADIOACTIVE ISOTOPES
REPRINTS
SENSE ORGANS
SITES
SPLEEN
SURFACES
Toxicology
TOXINS AND ANTITOXINS
TRYPSIN
VERO CELLS
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Summary:Mice were immunized with a cell line (Vero) that possesses a high number of membrane receptors for diphtheria toxin. Spleen cells from these mice were fused with SP2/0-Ag14 cells and two cell lines (1A2 and 2D2) isolated by screening for the ability of their secreted antibodies to inhibit binding of radiolabeled diphtheria toxin to Vero cells. These antibodies protected Vero cells from the inhibition of protein synthesis mediated by diphtheria toxin. The antibodies were purified, iodinated, and their binding characteristics investigated. At 4C, the association of 1A2 and 2D2 with Vero cells was saturable and indicated about 10(6) binding sites/cell. Diphtheria toxin did not inhibit the binding of either radiolabeled antibody. Monoclonal antibody 1A2 completely inhibited (125)I-2D2 binding and vice versa. Trypsin or phospholipase C treatment of Vero cells had no effect on the ability of the monoclonal antibodies to bind to the cells. These findings suggest that: (1) the two monoclonal antibodies recognize the same or closely related epitopes and (2) the antibodies bind a domain distinct from the toxin binding site or to a subcomponent of the diphtheria toxin receptor that is present at many other cell surface sites. These antibodies offer a powerful tool to study the structure, processing and mode of action of diphtheria toxin receptors. Reprints. Availability: Pub. in Toxicom, v27 n10 p1095-1104 1989.