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A sensitive chemiluminescence assay for pertussis toxin and for evaluation of cell-free pertussis toxoids
Authors:FF Craig  M Christodoulides  R Parton  DES Stewart-Tull  JH Freer  JM Lackie
Affiliation:Department of Microbiology, University of Glasgow, Glasgow, U.K.;Department of Cell Biology, University of Glasgow, Glasgow, U.K.
Abstract:Abstract 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 pertusis , 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.
Keywords:Bordetella pertussis            Chemiluminescence  Neutrophil  Pertussis toxin  Toxoid
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