ABSTRACT
Understanding of how the human pathogen Streptococcus pneumoniae perceives and responds to its environment in the host offers insight into the pathogenesis of disease caused by this important bacterium and the potential for improved interventions. A central role in this environmental response is played by two-component systems (TCSs), which both sense the environment and drive the cellular response. Molecular advances in the form of genome sequencing, signature-tagged mutagenesis, differential fluorescence induction and microarray analysis have yielded considerable progress in the study of these systems in S. pneumoniae. These recent advances are discussed here, focusing in particular on the role of TCSs in the virulence of S. pneumoniae.
Subject(s)
Bacterial Proteins/metabolism , Streptococcus pneumoniae/metabolism , Streptococcus pneumoniae/pathogenicity , Bacterial Proteins/genetics , Gene Expression Regulation, Bacterial , Genes, Bacterial/genetics , Streptococcus pneumoniae/genetics , Virulence/geneticsABSTRACT
Complement, which bridges innate and adaptive immune responses as well as humoral and cell-mediated immunity, is antiviral. Kaposi's sarcoma-associated herpesvirus (KSHV) encodes a lytic cycle protein called KSHV complement control protein (KCP) that inhibits activation of the complement cascade. It does so by regulating C3 convertases, accelerating their decay, and acting as a cofactor for factor I degradation of C4b and C3b, two components of the C3 and C5 convertases. These complement regulatory activities require the short consensus repeat (SCR) motifs, of which KCP has four (SCRs 1 to 4). We found that in addition to KCP being expressed on the surfaces of experimentally infected endothelial cells, it is associated with the envelope of purified KSHV virions, potentially protecting them from complement-mediated immunity. Furthermore, recombinant KCP binds heparin, an analogue of the known KSHV cell attachment receptor heparan sulfate, facilitating infection. Treating virus with an anti-KCP monoclonal antibody (MAb), BSF8, inhibited KSHV infection of cells by 35%. Epitope mapping of MAb BSF8 revealed that it binds within SCR domains 1 and 2, also the region of the protein involved in heparin binding. This MAb strongly inhibited classical C3 convertase decay acceleration by KCP and cofactor activity for C4b cleavage but not C3b cleavage. Our data suggest similar topological requirements for cell binding by KSHV, heparin binding, and regulation of C4b-containing C3 convertases but not for factor I-mediated cleavage of C3b. Importantly, they suggest KCP confers at least two functions on the virion: cell binding with concomitant infection and immune evasion.
Subject(s)
Complement System Proteins/physiology , Viral Proteins/chemistry , Animals , Antibodies, Monoclonal/immunology , Binding Sites , Cell Adhesion , Cells, Cultured , Endothelial Cells/metabolism , Humans , Mice , Mice, Inbred BALB C , Protein Structure, Tertiary , Recombinant Proteins/pharmacology , Viral Proteins/physiology , Virion/metabolismABSTRACT
IL-18, a multifunctional cytokine, has been shown to be involved in the immune response to numerous pathogens including several bacterial species. To study its role in infection by the Gram-positive bacterium Streptococcus pneumoniae, wild-type and IL-18 knockout BALB/c mice were compared in murine models of pneumococcal pneumonia, bacteraemia and nasopharyngeal colonization. The influence of IL-18 varied with the infection type, whereby it contributed to increased bacterial loads in pneumonia, reduced levels of colonization and had no effect on levels of bacteraemia following intravenous challenge. Likewise, the influence of IL-18 on pneumonia varied between two infecting pneumococcal strains. Comparison of these results with previous data also suggested that the influence of IL-18 in pneumococcal pneumonia differs with the mouse strain genetic background. Overall, these results demonstrate the complex influence of IL-18 in the response to the pneumococcus.
Subject(s)
Interleukin-18/genetics , Lung Diseases/immunology , Lung Diseases/microbiology , Pneumococcal Infections/immunology , Streptococcus pneumoniae , Animals , Disease Models, Animal , Interleukin-18/deficiency , Mice , Mice, KnockoutABSTRACT
Inflammation is a prominent feature of Streptococcus pneumoniae infection in both humans and animal models. Indeed, an intense host immune response to infection is thought to contribute significantly to the pathology of pneumococcal pneumonia and meningitis. Previously, induction of the inflammatory response following infection with S. pneumoniae has been attributed to certain cell wall constituents and the toxin pneumolysin. Here we present data implicating a putative zinc metalloprotease, ZmpB, as having a role in inflammation. Null mutations were created in the zmpB gene of the virulent serotype 2 strain D39 and analyzed in a murine model of infection. Isogenic mutants were attenuated in pneumonia and septicemia models of infection, as determined by levels of bacteremia and murine survival. Mutants were not attenuated in colonization of murine airways or lung tissue. Examination of cytokine profiles within the lung tissue revealed significantly lower levels of the proinflammatory cytokine tumor necrosis factor alpha following challenge with the Delta zmpB mutant (Delta 739). These data identify ZmpB as a novel virulence factor capable of inducing inflammation in the lower respiratory tract. The possibility that ZmpB was involved in inhibition of complement activity was examined, but the data indicated that ZmpB does not have a significant effect on this important host defense. The regulation of ZmpB by a two-component system (TCS09) located immediately upstream of the zmpB gene was examined. TCS09 was not required for the expression of zmpB during exponential growth in vitro.
