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In vitro and in vivo analyses of constitutive and in vivo-induced promoters in attenuated vaccine and vector strains of Vibrio cholerae.
John, M; Crean, T I; Calderwood, S B; Ryan, E T.
Afiliação
  • John M; Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
Infect Immun ; 68(3): 1171-5, 2000 Mar.
Article em En | MEDLINE | ID: mdl-10678922
The optimal promoter for in vivo expression of heterologous antigens by live, attenuated vaccine vector strains of Vibrio cholerae is unclear; in vitro analyses of promoter activity may not accurately predict expression of antigens in vivo. We therefore introduced plasmids expressing the B subunit of cholera toxin (CtxB) under the control of a number of promoters into V. cholerae vaccine strain Peru2. We evaluated the tac promoter, which is constitutively expressed in V. cholerae, as well as the in vivo-induced V. cholerae heat shock htpG promoter and the in vivo-induced V. cholerae iron-regulated irgA promoter. The functionality of all promoters was confirmed in vitro. In vitro antigenic expression was highest in vaccine strains expressing CtxB under the control of the tac promoter (2 to 5 microgram/ml/unit of optical density at 600 nm [OD(600)]) and, under low-iron conditions, in strains containing the irgA promoter (5 microgram/ml/OD(600)). We orally inoculated mice with the various vaccine strains and used anti-CtxB immune responses as a marker for in vivo expression of CtxB. The vaccine strain expressing CtxB under the control of the tac promoter elicited the most prominent specific anti-CtxB responses in vivo (serum immunoglobulin G [IgG], P
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Vibrio cholerae / Vacinas Bacterianas / Regiões Promotoras Genéticas Tipo de estudo: Prognostic_studies Limite: Animals Idioma: En Revista: Infect Immun Ano de publicação: 2000 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Vibrio cholerae / Vacinas Bacterianas / Regiões Promotoras Genéticas Tipo de estudo: Prognostic_studies Limite: Animals Idioma: En Revista: Infect Immun Ano de publicação: 2000 Tipo de documento: Article País de afiliação: Estados Unidos