• Korean Journal of Veterinary Research
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• v.39 no.2
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• pp.301-310
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• 1999

Anthrax caused by Bacillus anthracis is one of the most important zoonotic diseases. The bacterium produces several virulence factors. Of the factors, protective antigen (PA) of tripatite toxin has been identified as a central component in the pathogenesis of anthrax. However, precise roles of PA and other cellular components in the reaction with the target cells remain to be elucidated, especially in the initial stage of the disease. Three B anthracis antigens were prepared for investigation; PA, sonicated cellular antigens (S-Ag) and formalin-inactivaed whole cell antigens (W-Ag). PA was purified from culture supernatant of the bacterium using FPLC system with MonoQ. S-Ag and W-Ag were prepared by sonication and formalin inactivation of the cultured cells, respectively. Purity of the antigens was confirmed by SDS-PAGE and Western blot analysis. The roles of these antigens in the production of inflammatory mediators such as NO, IL-6 and $TNF{\alpha}$ from mouse peritoneal macrophages were investigated. PA alone did not induce the production of the inflammatory mediators while the other antigens, S-Ag and W-Ag, did in a dose and time dependent manner. These results suggested that in addition to major virulence factors, other cellular antigens are also involved in the initial stage of the disease by the induction of inflammatory mediators.

• Korean Journal of Microbiology
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• v.37 no.1
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• pp.21-27
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• 2001

Recently many investigators have devoted considerable attention to the production and purification of PA for antigens, and the preparation of new synthetic medium (RM medium) have solved to increase the yields of the PA but, the low sensitivity of the PA to detect B. anthracis infections has remained as a problem to be solved. This study was undertaken to evaluate the yields of the PA from culture filtrates of B. anthracis Sterne $34F_2$ strain in modified RM medium in which 10 g/l of $NaHCO_3$ and 10g/l of glucose were replaced by 8 g/l and 5 g/l, and the first purification step of PA from culture broth was used hydroxyapatite. The PA was purified by hydroxyapatite column chromatography, DEAE-Sepharose CL-4B column chromatography and Toyo-pearl gel filtration chromatography. The yield of PA from the modified RM medium, 8.6 mg/l.