A mutation in an ompR-like gene on a Legionella longbeachae serogroup 1 plasmid attenuates virulence

Examination of a panel of Legionella longbeachae serogroup 1 strains using a guinea pig model of virulence determined that this clonal species of Legionella shows a remarkable variation in symptoms and disease outcome (Doyle et al., Infect. Immun. 69, 5335-5344, 2001). The presence of plasmids was investigated, as plasmid encoded functions may contribute to the virulence of genetically similar strains. Partial sequence analysis of a large native plasmid (approximately 120 kb), designated pA5H5, from a highly virulent Australian isolate revealed a putative two-component regulatory system with inferred identity to the OmpR family of two-component transcriptional regulatory proteins and EnvZ sensor kinases. An isogenic mutant was constructed in the transcriptional regulatory gene, designated lrpR (L. longbeachae sg 1 regulatory protein) and this strain was tested in Acanthamoeba, U937 cells and in a guinea pig animal model. The mutant was reduced in intracellular multiplication within Acanthamoeba but not U937 macrophage-like cells. However, the lrpR mutant did appear reduced in invasion at the early stages of infection of U937 cells. The lrpR mutant was also attenuated for virulence in a guinea pig animal model of infection. The importance of plasmid-encoded functions for the pathogenicity of Legionella longbeachae serogroup 1 strains is discussed.