Subject(s)
Cholestasis, Intrahepatic/complications , Cholestasis, Intrahepatic/diagnosis , Pruritus/diagnosis , Pruritus/etiology , Adult , Cholagogues and Choleretics/therapeutic use , Cholestasis, Intrahepatic/drug therapy , Diagnosis, Differential , Female , Humans , Jaundice, Obstructive/complications , Jaundice, Obstructive/diagnosis , Liver Function Tests , Pemphigoid Gestationis/diagnosis , Pregnancy , Pregnancy Complications , Pruritus/drug therapy , Scabies/diagnosis , Skin Diseases, Papulosquamous/diagnosis , Skin Diseases, Papulosquamous/etiology , Ursodeoxycholic Acid/therapeutic useABSTRACT
This study examined the ability of the adenylate cyclase toxin (CyaA) of Bordetella pertussis to act as a mucosal adjuvant for other antigens when co-administered by the intranasal route in mice. Two forms of CyaA were used: the cell-invasive, enzymically active form and a cell-invasive toxin lacking adenylate cyclase enzymic activity (CyaA*). Co-administration intranasally (i/n) of CyaA or CyaA* with ovalbumin (Ova) significantly enhanced (P<0.05) anti-Ova IgG and IgA antibody responses in the serum and anti-Ova IgA responses in lung and nasal secretions compared to those generated by immunisation i/n with Ova alone. The effects were greater with CyaA*. Administration of CyaA* with Ova induced priming of Ova-specific T cells in vivo to a greater extent than that obtained after immunisation with Ova alone. Co-administration of CyaA or CyaA* with pertactin (Prn) significantly enhanced (P<0.05) the serum anti-Prn IgG responses and immunisation with Prn and CyaA* significantly increased the anti-Prn IgA responses in the lungs compared with responses after immunisation with Prn alone. Immunisation i/n with Prn alone partially protected mice (P<0.05) against challenge i/n with B. pertussis. Co-administration of CyaA or CyaA* with pertactin (Prn) significantly increased protection (P<0.05) against challenge compared to that obtained with Prn alone. These effects were particularly apparent with CyaA* as the adjuvant.
Subject(s)
Adenylate Cyclase Toxin/immunology , Adjuvants, Immunologic , Bordetella pertussis/immunology , Pertussis Vaccine/immunology , Whooping Cough/immunology , Adenylate Cyclase Toxin/administration & dosage , Administration, Intranasal , Animals , Antibodies, Bacterial/blood , Bacterial Outer Membrane Proteins/administration & dosage , Bacterial Outer Membrane Proteins/immunology , Bordetella pertussis/enzymology , Female , Immunization , Lung/immunology , Mice , Mice, Inbred BALB C , Nose/immunology , Ovalbumin/administration & dosage , Ovalbumin/immunology , Pertussis Vaccine/administration & dosage , T-Lymphocytes/immunology , Virulence Factors, Bordetella/administration & dosage , Virulence Factors, Bordetella/immunology , Whooping Cough/prevention & controlABSTRACT
Continuous recording of the activity of recombinant adenylate cyclase (CyaA) of Bordetella pertussis (EC ) by conductimetric determination of enzyme-coupled pyrophosphate cleavage has enabled us to define a number of novel features of the activation of this enzyme by calmodulin and establish conditions under which valid activation data can be obtained. Activation either in the presence or absence of calcium is characterized by a concentration-dependent lag phase. The rate of formation and breakdown of the activated complex can be determined from an analysis of the lag phase kinetics and is in good agreement with thermodynamic data obtained by measuring the dependence of activation on calmodulin concentration, which show that calcium increases k(on) by about 30-fold. The rate of breakdown of the activated complex, formed either in the presence or absence of calcium, has been determined by dilution experiments and has been shown to be independent of the presence of calcium. The coupled assay is established as a rapid, convenient and safe method which should be readily applicable to the continuous assays of most other enzymes that catalyze reactions in which inorganic pyrophosphate is liberated.
