Impact of the virulence-associated MAb3/1 epitope on the physicochemical surface properties of Legionella pneumophila sg1: An issue to explain infection potential?

The relationship between the presence/absence of the virulence-associated MAb3/1 epitope of sixteen Legionella pneumophila serogroup 1 strains and their respective surface physicochemical properties is evidenced from electrokinetic measurements (microelectrophoresis) performed as a function of KNO(3) electrolyte concentration (range 1-100mM, pH∼6.5). Among the bacteria selected, nine original strains constitute the Dresden reference panel and differ according to the presence/absence of the virulence-associated monoclonal antibody MAb3/1 of the O-specific chain of the lipopolysaccharides (LPS). Five isogenic Lens strains, also investigated in the current study, present the epitope MAb3/1 of their LPS and were involved to some extent in the outbreak that stroke the Nord Pas-de-Calais region (France) in 2004. All bacteria exhibit the typical electrokinetic features of soft (permeable) particles. On the basis of Ohshima's model, analysis of the electrophoretic mobility data allows evaluating the intraparticular flow penetration length 1/λ(0) and the (negative) volume charge density ρ(0) that both reflect the structure and chemical composition of the soft bacterial component. Our results show that the virulent MAb3/1 positive strains are characterized on average by 1/λ(0) and ǀρ(0)ǀ values that are about 1.5 times larger and 5 times lower, respectively, than those derived for lesser virulent (MAb3/1 negative) strains. In other words, on average the soft surface layer of MAb3/1 positive strains is significantly less charged and more permeable than those of MAb3/1 negative strains. The intimate correlation between virulence-associated MAb3/1 epitope and charge density carried by the bacterial envelop was further confirmed by lower 1/λ(0) and greater ǀρ(0)ǀ values for lag-1 mutant CS332 strain, lacking the MAb3/1 epitope, compared to the parental strain AM511. A closer inspection of the dispersion in 1/λ(0) and ǀρ(0)ǀ data over the ensemble of analysed bacteria together with the reported number of Legionnaires' disease cases they are responsible for, points out the charge density ǀρ(0)ǀ as the parameter that is most suitable for discriminating highly virulent (MAb3/1 positive) from less virulent (MAb3/1 negative) strains. Although short-range interaction determines infection process, our results suggest that the infection potential of Legionella pneumophila serogroup 1 may be also controlled significantly by non-specific long-range electrostatic repulsion the bacteria undergo when approaching negatively charged host cells to be infected.