FEMS Immunology and Medical Microbiology 9 (1994) 117-124 © 1994 Federation of European Microbiological Societies 0928-8244/94/$07.00 Published by Elsevier 117 FEMSIM 00414 Use of cyclodextrin as an agent to of Bordetella pertussis antigens induce excretion Daniela Hozbor, Maria E. Rodriguez and Osvaldo Yantorno * Centro de Investigacion y Desarrollo en Fermentaciones Industriales (CINDEFI), Facultad De Ciencias Exactas UNLP, Calle 47y 115 (1900), La Plata, Argentina (Received 10 January 1994; revision received 30 March 1994; accepted 12 April 1994) Abstract: This paper attempts to provide an explanation for the effect of cyclodextrin on the yield of Bordetella pertussis soluble antigens. It was demonstrated that the addition of cyclodextrin to the synthetic Stainer-Scholte liquid medium enhances the level of the intracellular form of adenylate cyclase (200 kDa) in the supernate. In addition to this effect, it has been reported that cyclodextrin also enhances the levels of two other extracellular proteins, pertussis toxin and filamentous hemagglutinin. As these antigens are structurally different, it seems that the effect of cyclodextrin is not specific. With the use of different buffer systems of well-known action on outer membrane stability it was possible to determine a relationship between the presence of cyclodextrin, destabilisation of the outer membrane and the release of proteins. It was determined that the cyclodextrin did not modify the fluidity of B. pertussis cells but produced a change of outer membrane permeability. Key words: Bordetella pertussis; Cyclodextrin; Outer membrane; Antigen release Introduction Among the virulence factors produced by Bor- detella pertussis, the ethiological agent of whoop- ing cough, adenylate cyclase (AC) has received considerable attention as a potential vaccine can- didate [1-5]. However, vaccine production based on this protein, using cultures of the virulent organism as the source of material, has been rather difficult because of the low yields of this protein secreted into culture fluids. The discovery that addition of 2,6-O-dimethyl-/3-cyclodextrin * Corresponding author. (Me/~CD) to the Stainer-Scholte (SS) [6] medium led to an enhanced level of AC, proved to be helpful [7]. Other authors have reported that cyclodextrin also increased the level of the other important soluble immunogens of B. pertussis such as pertussis toxin (PT) and filamentous hemagglutinin (FHA) [8-11]. However, the way which this compound enhances the yield of extra- cellular antigens, and in particular that of AC, remains to be elucidated. For the development of an acellular vaccine such knowledge is important, from an economic point of view, because it could contribute to the search for other, less expensive substances which could evoke a similar effect. In a previous paper [12] we showed that cy- SSDI 0928-8244(94)00022-L Downloaded from https://academic.oup.com/femspd/article-abstract/9/2/117/518750 by guest on 22 May 2020