Detection of antibodies inhibiting the ADP-ribosyltransferase activity of pertussis toxin in human serum.
Bordetella pertussis produces a protein virulence factor termed pertussis toxin. Many candidate pertussis vaccines are based on the rationale that an immune response that neutralizes the virulence activities of this toxin, which are thought to arise from its catalytic ADP-ribosyltransferase activity, would be beneficial. The report describes two methods that quantify the inhibition of this activity by human serum. One, termed a direct assay, involves an initial incubation of toxin with serum, a second incubation that activates the toxin, and a third incubation that measures the ADP-ribosyltransferase activity of the mixture. The other assay, termed a plate assay, involves immobilization of the toxin, exposure of the immobilized toxin to serum and washing of the plate, and then activation and assay of the toxin's ADP-ribosyltransferase activity. The plate assay may be more selective than the direct assay in terms of identifying antibodies that neutralize the toxin in vivo. Sera from controls, selected patients presenting with cough, and vaccinated infants were first analyzed by the direct assay. In contrast to sera from controls, sera from several of the patients and vaccinated infants strongly inhibited activity. Dose-response curves of inhibition were determined for samples from three vaccinated infants by both the direct and plate assays. One of the samples had a dose-response curve of a different shape and thus differed not only in titer but also in functional characteristics. A comparison of inhibition of ADP-ribosyltransferase activity and neutralization in a CHO cell assay indicated that there was incomplete agreement between the two assays. Taken together, these results indicate that measurement of inhibition of ADP-ribosyltransferase activity by human serum is practical and may be useful in the evaluation of responses to pertussis vaccines.
Kaslow
H R
HR
Department of Physiology and Biophysics, University of Southern California School of Medicine, Los Angeles 90033.
Platler
B W
BW
Blumberg
D A
DA
Cherry
J D
JD
D000293:Adolescent; D000328:Adult; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood* / Q000276:immunology; D016466:CHO Cells; D006224:Cricetinae; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D008875:Middle Aged; D037342:Pertussis Toxin; D010567:Pertussis Vaccine / Q000276:immunology; D011065:Poly(ADP-ribose) Polymerases / Q000097:blood / Q000276:immunology*; D010566:Virulence Factors, Bordetella / Q000097:blood / Q000276:immunology*
D000907:Antibodies, Bacterial; D010567:Pertussis Vaccine; D010566:Virulence Factors, Bordetella; D011065:Poly(ADP-ribose) Polymerases; D037342:Pertussis Toxin