Outer membrane protein A (OmpA) contributes to serum resistance and pathogenicity of Escherichia coli K-1

Abstract

We examined whether outer membrane protein A (OmpA) contributes to gram-negative pathogenesis by determining the effect of mutagenesis of ompA in a virulent Escherichia coli K-1 isolate. An OmpA mutant was generated by insertion of the transposon TnphoA, which was genetically modified to increase the efficiency of its delivery by conjugation. The mutant was less virulent than its parent strain in two models of E. coli K-1 infection. Equal inocula of the OmpA+ and OmpA- strains fed to neonatal rats resulted in a sevenfold-greater incidence of bacteremia at 72 h from the OmpA+ strain. The lethal effect of the OmpA- mutant was significantly less than that of the OmpA+ parent strain when inoculated onto the chorioallantoic membrane of 10-day embryonated chick eggs. There was, however, no difference between strains in growth characteristics under physiologic conditions, either in rat serum or in unembryonated chick eggs. In the presence of a 10-day chick embryo, there was a 10-fold increase in the survival and growth of the OmpA+ strain. Correction of the mutation in ompA with an E. coli K-12 ompA gene restored a level of virulence equivalent to that of the parent strain. The ompA mutant was more sensitive to the bactericidal effect of pooled human serum by the classical pathway of complement activation. These results suggest that OmpA contributes to E. coli K-1 pathogenesis by a mechanism which may involve increased serum resistance.