Superantigen genes are more important than the emm type for the invasiveness of group A Streptococcus infection.

BACKGROUND: In this study, we examined the role of superantigen genes and emm genotypes of clinical Streptococcus pyogenes isolates collected in Germany between 1997 and 2003. METHODS: Multiplex polymerase chain reaction for all 11 currently known superantigen genes and sequencing for emm types were used. RESULTS: Using a 2-step explorative data analysis procedure, we found that after combined analysis of superantigen genes and emm types, only the superantigen genes spea1-spea3, spem, and spea4 have a predictive value for invasiveness, with odds ratios of 7.992, 3.209, and 2.323, respectively. The predictive value for invasiveness of emm1 was lost after combined analysis because of the association between emm type and the highly predictive superantigen genes. On the other hand, presence of the superantigen gene ssa and of emm77 are predictors of noninvasiveness, with odds ratios of 0.370 and 0.271, respectively. CONCLUSIONS: These results indicate that the presence of superantigen genes is more important for the invasiveness of group A Streptococcus infection than emm type and may be the connection between the high-risk HLA type of the host and the pathogen. Furthermore, we found a very clear correlation between the presence of the genes spea1-spea3 and the presence of the gene emm1, which indicates that the relationship between emm1 and invasiveness is based on the superantigen gene profile. Our data suggest that the superantigen gene profile is of high importance for the clinical outcome of group A Streptococcus infections.

A new closed-tube multiplex real-time PCR to detect eleven superantigens of Streptococcus pyogenes identifies a strain without superantigen activity.

Superantigens (SAgs) are very potent microbial toxins that are involved in severe diseases such as necrotizing fasciitis and toxic shock syndrome. There are currently 11 different SAgs that have been identified from Streptococcus pyogenes. In the present study, two sets of multiplex PCRs were developed for detection of these 11 SAg genes. The first group comprises spea1-3+5, spec, speg, spej, spek, and spel. The second group consists of spea1-4, speh, spei, spem, ssa, and smez. The presence of Streptococcus pyogenes SAg genes can be immediately identified using a real-time method with SYBR-Green, thus providing an excellent tool in clinical diagnostics. After testing more than 300 clinical isolates, we identified one strain without any SAg gene. This finding contrasts with previous reports describing SAg genes located on every Streptococcus pyogenes genome. This SAg gene-negative strain also did not show any mitogenic activity. It is hypothesized that clinical isolates from patients may overrepresent bacterial strains with pathogenic factors, such as SAgs.

Mycoplasma arthritidis-derived superantigen (MAM) displays DNase activity.

Bacterial superantigens are potent stimulators of the immune system. In this study, we expressed recombinant superantigens, which were then affinity purified and used for growth curves and DNase activity assays. Overexpression of Mycoplasma arthritidis-derived superantigen in Escherichia coli reduced bacterial growth. This is unique, as staphylococcal enterotoxin A and toxic shock syndrome toxin-1, expressed in the same vector system, showed no growth impairment. The observed growth inhibition was caused by the DNase activity of recombinant M. arthritidis-derived superantigen, thus describing the first superantigen showing enzymatic activity, which may be a result of the separate evolution of this toxin.

The superantigen staphylococcal enterotoxin A (SEA) and monoclonal antibody L243 share a common epitope but differ in their ability to induce apoptosis via MHC-II.

Crosslinking of MHC class II (MHC-II) molecules by antibodies or by superantigens (SAg) induces a variety of functional responses in the antigen presenting cell. We were able to allocate K39 as the residue that is essential for binding of antibody L243 to the alpha chain of HLA-DR. K39 is also essential for binding of staphylococcal enterotoxin A (SEA). However, the functional responses of the two ligands differ considerably exemplified by the ability of L243 to induce apoptosis in monocytic cells and in B cells, whereas SEA is unable to activate the apoptosis pathway. Despite the differences in functional responses, both ligands induce cell aggregation in MonoMac-1 cells. The SEA molecule with its two different binding sites associates one MHC alpha chain with one beta chain as opposed to two alpha chains that are brought into close proximity by the two identical antigen binding sites of L243. We therefore conclude that the spatial orientation of dimerized MHC-II and their associated proteins is an important factor for the nature of the transduced signal and consequently the outcome of functional responses.