Abstract
We describe the development of plasmid vectors carrying the expression sites, an hlyA cassette and the secretion genes of Escherichia coli hemolysin. These allow the synthesis and secretion of heterologous microbial antigens in E. coli and attenuated Salmonella aroA strains. Genes or gene fragments encoding microbial antigens are inserted in-frame into a residual part of the hlyA gene which essentially encodes the HlyA secretion signal (HlyAs). In general, the fused genes, carrying the hlyAs sequence at the 3' terminus, are efficiently expressed, and the synthesized antigens are secreted into the culture supernatant of the producing strain. Attenuated Salmonella strains synthesizing either HlyAs-fused listeriolysin or p60 of Listeria monocytogenes were constructed by this procedure and shown to provide protective immunity against L. monocytogenes in mice. The most effective protection was obtained when these microbial antigens were secreted by the attenuated Salmonella strains. We further present new approaches which may allow the application of this antigen-delivery system to any microbial antigen.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antigens, Bacterial / administration & dosage*
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Antigens, Bacterial / biosynthesis
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Antigens, Bacterial / genetics
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Antigens, Bacterial / therapeutic use
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Bacterial Proteins / administration & dosage*
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism
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Bacterial Proteins / physiology*
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Bacterial Toxins*
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Bacterial Vaccines / administration & dosage
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Bacterial Vaccines / genetics
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Bacterial Vaccines / therapeutic use
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Escherichia coli Proteins*
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Genetic Vectors
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Heat-Shock Proteins / administration & dosage*
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Heat-Shock Proteins / genetics
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Heat-Shock Proteins / metabolism
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Hemolysin Proteins / genetics
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Hemolysin Proteins / physiology*
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Listeriosis / prevention & control
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Mice
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Plasmids / genetics
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Recombinant Fusion Proteins / administration & dosage
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Recombinant Fusion Proteins / metabolism
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Recombinant Fusion Proteins / therapeutic use
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Salmonella / genetics
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Vaccination
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Vaccines, Synthetic / administration & dosage
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Vaccines, Synthetic / genetics
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Vaccines, Synthetic / therapeutic use
Substances
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60 kDa protein, Listeria monocytogenes
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Antigens, Bacterial
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Bacterial Proteins
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Bacterial Toxins
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Bacterial Vaccines
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Escherichia coli Proteins
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Heat-Shock Proteins
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Hemolysin Proteins
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Hlya protein, E coli
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Recombinant Fusion Proteins
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Vaccines, Synthetic
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hlyA protein, Listeria monocytogenes