well-characterised virulence factors and discusssome of the newer data relating to host responseand the role of two-component transcriptionalregulators (TCS) in virulence. A summary of some of the putative virulence factors of thepneumococcus is shown in
Much work has been carried out on the roleof this protein in infection models. The genewas originally cloned in 1983 [6, 7] and theavailability of this gene allowed the construc-tion of de
ned pneumolysin-negative knock-outs and isogenic replacement mutants [8
10].These mutants have been used in several labo-ratories to probe the role of the toxin. The toxinitself has been the subject of a detailed structurefunction analysis and a three-dimensional struc-ture has been proposed based on the structureof the related toxin perfringolysin O. Functionaldomains have been mapped onto this structure. By altering these domains andre-introducing the gene into the pneumococcusit has been possible to probe the role of indi-vidual amino acids in the pathogenesis of infec-tion . A summary of the effects of the muta-tions and the possible role of pneumolysin isgiven in
. Pneumolysin has at least twobiological activities that contribute to virulencein animal models: i) lytic activity and ii) abilityto activate complement. However, introductionofpoint mutations,which block these two activi-ties, results in an organism that is still morevirulent than the pneumolysin-negative mutant. This suggests that there is another activityof the toxin which contributes to virulence.
4. Surface proteins
The surface of the pneumococcus is decoratedwith a range of surface proteins that may play arole in pathogenesis. These proteins may beeither anchored to the cell wall via the Gram-positive attachment motif (LPXTG) or may benoncovalently attached via an interaction withcholine (choline-binding proteins, Cbps). One of the neuraminidase enzymes and hyaluronidaseproduced by the pneumococcus spend part of their time as cell surface proteins as judged bythe presence of the LPXTG motif and cell locali-sation experiments . Choline-binding pro-teins share common repeat elements associatedwith this property and include PspA, LytA, andCbpA which will be described here.
4.1. Pneumococcal surface protein A (PspA)
The mechanism of action of PspA is not fullyunderstood, but the protein is present on thesurface of all pneumococci and is required forfull viulence [14, 15]. It has been reported thatPspA is a lactoferrin-binding protein . PspAalso inhibits complement activation by
. The amino-terminal half of the protein contains a highly charged coil
coil domain and it is variations in this sequencethat generate the heterogeneity observed in thismolecule. The region still contains sufficientconserved epitopes to allow vaccination with asingle PspA type to confer protective immunityto a range of other PspA types .
4.2. Choline-binding protein (CbpA)
CbpA is present on the surface of the pneu-mococcus and reacts with pooled convalescent
. Some of the virulence factors produced by
414 T.J. Mitchell / Res. Microbiol. 151 (2000) 413