Helicobacter pylori is a major cause of gastric-associated diseases. To evaluate the efficacy of a possible vaccine antigen against H. pylori infection, the chimaeric construct adhesin–CTXA2B, derived from H. pylori adhesin genetically coupled to cholera toxin (CTX) subunits A2 and B (CTXA2B), was expressed in Escherichia coli as an insoluble recombinant chimaeric protein. The protein was then purified by denaturation, renaturation and size-exclusion chromatography. The composition of purified adhesin–CTXA2B was verified by SDS/PAGE and Western blotting with antibodies to antigenic components of adhesin and CTXB, and confirmed as a chimaeric protein with GM1-ganglioside binding activity and adhesin epitopes by a GM1-ELISA developed using antibodies to adhesin. Oral immunization of mice with adhesin–CTXA2B induced higher levels of mucosal IgA and serum IgG antibodies to H. pylori adhesin and to CTXB than in mice immunized with adhesin or CTXA2B alone. Adhesin–CTXA2B was also demonstrated to be a potential protective antigen in a mouse model of H. pylori infection. The immunization of mice with adhesin–CTXA2B protected 62.5% of mice infected with H. pylori SS1 strain, whereas adhesin immunization was not able to confer protection to mice. This protection may be correlated with high levels of mucosal IgA and serum IgG antibodies against H. pylori adhesin. Taken together, the results indicate that the genetically linked CTXA2B acts as a useful mucosal adjuvant, and that the adhesin–CTXA2B chimaeric protein could be a potential component in future H. pylori vaccine development.
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Research Article|
February 02 2001
Peroral immunization with Helicobacter pylori adhesin protein genetically linked to cholera toxin A2B subunits
Byung Oh KIM;
Byung Oh KIM
*School of Pharmacy, Sung Kyun Kwan University, Suwon, 440-746, Kyunggi-Do, South Korea
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Sung Seup SHIN;
Sung Seup SHIN
†R&D Center, Daewoong Pharmaceutical Co., Ltd., Sungnam, 462-120, Kyunggi-Do, South Korea
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Young Hyo YOO;
Young Hyo YOO
†R&D Center, Daewoong Pharmaceutical Co., Ltd., Sungnam, 462-120, Kyunggi-Do, South Korea
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Suhkneung PYO
*School of Pharmacy, Sung Kyun Kwan University, Suwon, 440-746, Kyunggi-Do, South Korea
Correspondence: Dr Suhkneung Pyo (e-mail [email protected]).
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Publisher: Portland Press Ltd
Received:
May 26 2000
Revision Received:
October 09 2000
Accepted:
November 24 2000
Online ISSN: 1470-8736
Print ISSN: 0143-5221
The Biochemical Society and the Medical Research Society © 2001
2001
Clin Sci (Lond) (2001) 100 (3): 291–298.
Article history
Received:
May 26 2000
Revision Received:
October 09 2000
Accepted:
November 24 2000
Citation
Byung Oh KIM, Sung Seup SHIN, Young Hyo YOO, Suhkneung PYO; Peroral immunization with Helicobacter pylori adhesin protein genetically linked to cholera toxin A2B subunits. Clin Sci (Lond) 1 March 2001; 100 (3): 291–298. doi: https://doi.org/10.1042/cs1000291
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