Diphtheria toxoid, the principle component of diphtheria vaccines, is prepared by inactivating diphtheria toxin with formaldehyde and glycine. The treatment introduces intramolecular cross-links and intermolecular formaldehyde/glycine adducts in diphtheria toxin. The purpose of the present study was to elucidate the nature and location of formaldehyde-induced modifications at two functional sit...

We report here the synthesis and properties of a hybrid toxin prepared by covalently coupling diphtheria toxin to transferrin. The purified material contained two major hybrid protein species and was highly cytotoxic to mouse LMTK- cells in culture, reducing protein synthesis by 50% in 24 h at a concentration of 1 ng/ml. Cytotoxic activity was completely abolished in the presence of exogenous t...

THE RESULTS OBTAINED IN THE PRECEDING EXPERIMENTS MAY BE BRIEFLY SUMMARIZED AS FOLLOWS: 1. Nucleohiston does not protect against a separate and subcutaneous injection of tetanus toxin, diphtheria toxin, hog-cholera bacillus, or anthrax bacillus. 2. Mixtures of nucleohiston and either tetanus toxin or diphtheria toxin lose their specific toxic action-the latter much more rapidly than the former....

A reversed passive latex agglutination (RPLA) assay for determining the toxigenicity of Corynebacterium diphtheriae is presented. Rabbit antitoxin antiserum was raised by using commercially available diphtheria toxoid. This antiserum reacted with the diphtheria toxin when the culture supernatant was assayed by Western blotting, and it did not cross-react with other extracellular antigens. Affin...

A nonspecific binding of antibodies to diphtheria toxin, especially in adult serum samples, was observed in our diphtheria-tetanus-pertussis multiplex immunoassay (DTaP4 MIA). This can be significantly reduced by the use of diphtheria toxoid, achieving a good correlation with the Vero cell neutralization test and the toxin binding inhibition assay.

At neutral pH, NH4Cl and chloroquine protected cells against diphtheria toxin. A brief exposure of the cells to low pH (4.5-5.5) at 37 degrees completely abolished this protection. When, to cells preincubated with diphtheria toxin and NH4Cl, neutralizing amounts of anti-diphtheria toxin were added before the pH was lowered, the toxic effect was considerably reduced, but it was not completely ab...

A method for the screening of tetanus and diphtheria antibodies in serum using anatoxin (inactivated toxin) instead of toxin was developed as an alternative to the in vivo toxin neutralization assay based on the toxin-binding inhibition test (TOBI test). In this study, the serum titers (values between 1.0 and 19.5 IU) measured by a modified TOBI test (Modi-TOBI test) and toxin neutralization as...

The biochemical and biologic properties of a purified disulfide conjugate of diphtheria toxin fragment A and human placental lactogen (toxin A-hPL) have been studied by (a) assaying the ADP-ribosyltransferase activity of the intact conjugate, (b) assaying the binding of the intact conjugate to mammary gland plasma membrane lactogenic receptors, and (c) assaying the effect of the conjugate on th...

We describe here three different hamster cell mutants which are resistant to diphtheria toxin and which provide models for investigating some of the functions required by the toxin inactivates elongation factor 2 (EF-2). Cell-free extracts from mutants Dtx(r)-3 was codominant. The evidence suggests that the codominant phenotype is the result of a mutation in a gene coding for EF-2. The recessiv...

Chinese hamster ovary (CHO) cell cultures in microtiter wells are sensitive to growth inhibition and killing by picogram quantities of diphtheria toxin. In the absence of biologically active toxin, the CHO cell culture produces sufficient acidic metabolites to change the phenol red pH indicator from pink to yellow within 56 h. In the presence of 10 pg of toxin per well, growth inhibition can be...