Outer membrane protein PhoE as a carrier for the exposure of foreign antigenic determinants at the bacterial cell surface |
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Authors: | Marja Agterberg Jan Tommassen |
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Affiliation: | (1) Institute of Molecular Biology and Medical Biotechnology, University of Utrecht, Padualaan 8, 3584-CH Utrecht, The Netherlands;(2) Department of Molecular Cell Biology, University of Utrecht, Padualaan 8, 3584-CH Utrecht, The Netherlands;(3) Present address: Department of Medical Chemistry, Free University of Amsterdam, van der Boechorststraat 7, 1007-MC Amsterdam, The Netherlands |
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Abstract: | PhoE protein is an abundant outer membrane protein of theEscherichia coli K-12 outer membrane. This protein can be used as an exposure system to produce foreign antigenic determinants and for their transport to the bacterial cell surface. The system is very flexible, since insertions varying in length and nature could be made in different cell surface-exposed regions of PhoE, without interfering with the assembly process of the mutant proteins into the outer membrane. Two antigenic determinants of the structural VP1 protein of foot-and-mouth disease virus were inserted in different combinations in four cell surface-exposed regions of PhoE. The epitopes were exposed at the bacterial cell surface and they keep their antigenic and immunogenic properties in this PhoE-associated conformation. Immunization of guinea pigs with one hybrid protein, containing a combination of the two epitopes inserted in the fourth exposed region, resulted in complete protection against challenge with the virus. A T-cell epitope of the 65 kDa heat shock protein ofMycobacterium tuberculosis was inserted in the fourth exposed region of PhoE and in vitro proliferation of two T-cell specific clones was demonstrated. Thus, the PhoE exposure system has been shown to be suitable for presentation of both B-cell and T-cell determinants to the immune system. Furthermore, good expression of the hybrid protein in attenuatedSalmonella strains, which can be used as live oral vaccines, was shown.Paper awarded the Kluyver Prize 1990 by the Netherlands Society of Microbiology to Dr. M.C. Agterberg |
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Keywords: | exposure system cell surface immunization immunogenicity live oral vaccine PhoE protein |
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