title: ADP-ribosylation activity in pertussis vaccines and its relationship to the in vivo histamine-sensitisation test.
abstract: Pertussis toxin (PTx) is a major virulence factor produced by Bordetella pertussis. In its detoxified form (PTd), it is an important component of acellular pertussis vaccines although some residual PTx activity may likely be present because of the limitations of the detoxification processes used. Furthermore, different detoxification procedures have been shown to result in different amino acid side-chain modifications for the resulting PTd. The histamine-sensitisation test (HIST) in mice is currently used for the safety testing of these vaccines. However, an alternative test is needed because of large assay variability and ethical concerns. The ADP-ribosylation enzyme activity of PTx is thought to be the major factor responsible for the histamine-sensitising activity detected in vivo. In the present study, the ADP-ribosylation activity in different acellular pertussis-based combination vaccine formulations was measured and compared with reactivity in the HIST. The results indicated that different products showed differences in ADP-ribosylation activity and a level which would be significant in relation to the reactivity seen in the HIST could not be defined, except for vaccines that contain genetically detoxified PTx, which do not have enzymatic activity nor in vivo toxicity. Different detoxification procedures as well as formulation factors could contribute to this variation. Relying solely on the residual enzyme activity of PTx in vaccines containing chemically detoxified PTd may not fully reflect the in vivo reactivity observed by the HIST. Refinement of the in vitro test to include a step which monitors the B-subunit activity of PTx may provide a better correlation with the in vivo HIST.
authors.affiliation: Division of Bacteriology, National Institute for Biological Standards and Control, Potters Bar, Hertfordshire, EN6 3QG, UK.
authors.forename: S R
authors.initials: SR
authors.lastname: Gomez
authors.forename: C-T
authors.initials: CT
authors.lastname: Yuen
authors.forename: C
authors.initials: C
authors.lastname: Asokanathan
authors.forename: A
authors.initials: A
authors.lastname: Douglas-Bardsley
authors.forename: M J
authors.initials: MJ
authors.lastname: Corbel
authors.forename: J G
authors.initials: JG
authors.lastname: Coote
authors.forename: R
authors.initials: R
authors.lastname: Parton
authors.forename: D K L
authors.initials: DK
authors.lastname: Xing
mesh_terms: D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism*; D006632:Histamine / Q000494:pharmacology*; D051379:Mice; D009243:NAD / Q000378:metabolism; D037342:Pertussis Toxin / Q000032:analysis*; D010567:Pertussis Vaccine / Q000737:chemistry* / Q000494:pharmacology; D022282:Vaccines, Acellular / Q000737:chemistry
chemical_list: D010567:Pertussis Vaccine; D022282:Vaccines, Acellular; D009243:NAD; D000246:Adenosine Diphosphate Ribose; D006632:Histamine; D037342:Pertussis Toxin; D019206:GTP-Binding Protein alpha Subunits, Gi-Go