GENOTYPIC AND PHENOTYPIC CHARACTERIZATION OF AN AROD DELETION-ATTENUATED ESCHERICHIA-COLI K12-SHIGELLA-FLEXNERI HYBRID VACCINE EXPRESSING SHIGELLA-FLEXNERI 2A-SOMATIC ANTIGEN

The construction and characterization of EcSf2a-2, an aroD-deleted Escherichia coli-Shigella hybrid vaccine carrying chromosomal and plasmid genes from Shigella flexneri and expressing S. flexneri 2a somatic antigen in association with E. coli K12 core are described. Expression of hybrid lipopolysaccharide and deletion of aroD resulted in the attenuation of phenotypic characteristics associated with pathogenicity. The addition of an aroD deletion results in a requirement for an aromatic precursor of para-aminobenzoic acid (PABA), an essential bacterial metabolite not present in mammalian tissues. The biosynthesis of hybrid somatic antigen prevents expression of a Sereny-positive reaction by invasive bacteria capable of expressing a plaque-positive phenotype. A functional kcpA gene is required for expression of the plaque-positive phenotype. The presence of an aroD deletion does not interfere with expression of an invasive phenotype; however, in bacteria containing afunctional kcpA gene, replication and spread by invading bacteria are limited, preventing development of the plaque-positive phenotype.