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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 23 (1999), S. 0 
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Bordetella pertussis produces a cell-invasive adenylate cyclase toxin which is synthesised from the cyaA gene as an inactive protoxin that is post-translationally activated by the product of the cyaC gene. Purified active and inactive CyaA proteins were prepared from B. pertussis or from recombinant Escherichia coli expressing both cyaA and cyaC genes or the cyaA gene alone, respectively. In addition, a hybrid toxin (Hyb2) in which an internal region of CyaA had been replaced with the analogous region from the leukotoxin (LktA) of Pasteurella haemolytica, and which had low cell-invasive activity, was also prepared from E. coli expressing the cyaC gene. The CyaA preparations showed no evidence of toxicity in a mouse weight-gain test. Active toxin preparations were protective in mice against intranasal challenge with wild-type B. pertussis, as evidenced by lung:body weight ratios and bacterial numbers in the lungs, which were comparable to those in mice given whole-cell DPT vaccine. Hyb2 was not as protective as active CyaA and inactive CyaA preparations were not protective. Active CyaA, when co-administered with ovalbumin (OA), had a marked adjuvant effect on the anti-OA IgG antibody response which was not as apparent with inactive CyaA preparations. Similarly, active CyaA stimulated a greater anti-CyaA response than the inactive form.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The leukotoxin (LktA) of Pasteurella haemolytica is active only against cells of ruminant origin. It is synthesised as an inactive protoxin encoded by the lktA gene and post-translationally modified to the active toxin by the product of the lktC gene. The LktA and LktC proteins were expressed separately in Escherichia coli and partially purified. Active LktA was produced in vitro in the presence of LktC and acyl-acyl carrier protein (ACP) charged separately in vitro with a fatty-acyl group. The toxic activity and target cell specificity of LktA and adenylate cyclase toxin (CyaA), a toxin active against a wide variety of mammalian cells, were investigated after activation with ACP charged with different fatty acids. Palmitoyl-ACP produced the most active toxin in both cases and, although other fatty acids were also effective, the fatty acid preference was the same for the in vitro activation of both toxins. Activated LktA remained ruminant cell-specific whichever acyl group was used to acylate the A protoxin.
    Type of Medium: Electronic Resource
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