A sensitive chemiluminescence assay for pertussis toxin and for evaluation of cell-free pertussis toxoids.
Pertussis toxin (PT) inhibited luminol-enhanced chemiluminescence induced in rabbit peritoneal neutrophils by N'-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) at doses as low as 0.8 ng.ml-1, even in the presence of a 10-fold higher concentration of filamentous haemagglutinin (FHA). A cell-free extract of Bordetella pertussis, containing predominantly PT and FHA, suppressed the neutrophil response to fMLP. After toxoiding with carbodiimide, the inhibitory activity of the extract was abolished and an enhancement of neutrophil chemiluminescence was observed due to FHA activity. Abrogation of the chemiluminescent response of neutrophils to fMLP is proposed as a sensitive, in vitro assay for pT, and may be useful for monitoring the residual toxin activity in pertussis toxoids and for determining the anti-toxic effects of anti-PT antibodies.
Craig
F F
FF
Department of Microbiology, University of Glasgow, U.K.
Christodoulides
M
M
Parton
R
R
Stewart-Tull
D E
DE
Freer
J H
JH
Lackie
J M
JM
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D006388:Hemagglutinins / Q000494:pharmacology; D066298:In Vitro Techniques; D008163:Luminescent Measurements; D008165:Luminol; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects; D037342:Pertussis Toxin; D011817:Rabbits; D014121:Toxoids / Q000032:analysis* / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000032:analysis* / Q000494:pharmacology
D006388:Hemagglutinins; D014121:Toxoids; D010566:Virulence Factors, Bordetella; C051093:pertussis toxoid; D009240:N-Formylmethionine Leucyl-Phenylalanine; D008165:Luminol; D037342:Pertussis Toxin