AI-2 quorum sensing-induced galactose metabolism activation in Streptococcus suis enhances capsular polysaccharide-associated virulence.

Bacteria utilize intercellular communication to orchestrate essential cellular processes, adapt to environmental changes, develop antibiotic tolerance, and enhance virulence. This communication, known as quorum sensing (QS), is mediated by the exchange of small signalling molecules called autoinducers. AI-2 QS, regulated by the metabolic enzyme LuxS (S-ribosylhomocysteine lyase), acts as a universal intercellular communication mechanism across gram-positive and gram-negative bacteria and is crucial for diverse bacterial processes. In this study, we demonstrated that in Streptococcus suis (S. suis), a notable zoonotic pathogen, AI-2 QS enhances galactose utilization, upregulates the Leloir pathway for capsular polysaccharide (CPS) precursor production, and boosts CPS synthesis, leading to increased resistance to macrophage phagocytosis. Additionally, our molecular docking and dynamics simulations suggest that, similar to S. pneumoniae, FruA, a fructose-specific phosphoenolpyruvate phosphotransferase system prevalent in gram-positive pathogens, may also function as an AI-2 membrane surface receptor in S. suis. In conclusion, our study demonstrated the significance of AI-2 in the synthesis of galactose metabolism-dependent CPS in S. suis. Additionally, we conducted a preliminary analysis of the potential role of FruA as a membrane surface receptor for S. suis AI-2.

Structure and Signal Regulation Mechanism of Interspecies and Interkingdom Quorum Sensing System Receptors.

Quorum sensing (QS) is a signaling mechanism for cell-to-cell communication between bacteria, fungi, and even eukaryotic hosts such as plant and animal cells. Bacteria in real life do not exist as isolated organisms but are found in complex, dynamic, and microecological environments. The study of interspecies QS and interkingdom QS is a valuable approach for exploring bacteria-bacteria interactions and bacteria-host interaction mechanisms and has received considerable attention from researchers. The correct combination of QS signals and receptors is key to initiating the QS process. Compared with intraspecies QS, the signal regulation mechanism of interspecies QS and interkingdom QS is often more complicated, and the distribution of receptors is relatively wide. The present review focuses on the latest progress with respect to the distribution, structure, and signal transduction of interspecies and interkingdom QS receptors and provides a guide for the investigation of new QS receptors in the future.

Paeoniflorin reduce luxS/AI-2 system-controlled biofilm formation and virulence in Streptococcus suis.

Streptococcus suis (S. suis), more specifically serotype 2, is a bacterial pathogen that threatens the lives of pigs and humans. Like many other pathogens, S. suis exhibits quorum sensing (QS) system-controlled virulence factors, such as biofilm formation that complicates treatment. Therefore, impairing the QS involving LuxS/AI-2 cycle in S. suis, may be a promising alternative strategy for overcoming S. suis infections. In this study, we investigated paeoniflorin (PF), a monoterpenoid glycoside compound extracted from peony, as an inhibitor of S. suis LuxS/AI-2 system. At a sub-minimal inhibitory concentration (MIC) (1/16 MIC; 25 µg/ml), PF significantly reduced biofilm formation by S. suis through inhibition of extracellular polysaccharide (EPS) production, without affecting bacterial growth. Moreover, evidence was brought that PF reduces AI-2 activity in S. suis biofilm. Molecular docking indicated that LuxS may be the target of PF. Monitoring LuxS enzymatic activity confirmed that PF had a partial inhibitory effect. Finally, we showed that the use of PF in a mouse model can relieve S. suis infections. This study highlighted the anti-biofilm potential of PF against S. suis, and brought evidence that it may as an inhibitor of the LuxS/AI-2 system to prevent S. suis biofilm-related infections. PF can thus be used as a new type of natural biofilm inhibitor for clinical application.

Evaluation of immune effect of Streptococcus suis biofilm-associated protein PDH.

As a zoonotic pathogen, Streptococcus suis(S. suis) takes pigs as the main host and is mainly colonizes in the upper respiratory tract and tonsil of pigs, causing septicemia, endocarditis and meningitis in pigs. Pyruvate dehydrogenase (PDH) is an enzyme that catalyzes the conversion of pyruvate to acetyl-CoA. As an immunogenic membrane-associated protein in S. suis, it has been found to be closely related to the formation of biofilm. In this study, the recombinant PDH (rPDH) of S. suis ZY05719 (serotype 2) was expressed and purified in E. coli by His affinity chromatography. Western blotting analysis showed that there was a strong specific reaction between PDH protein and PDH antiserum. Mice were immunized with recombinant PDH and inactivated bacteria, and the relative survival rates were 70 % and 60 %, respectively. In addition, mice immunized with PDH caused high levels of antibodies and high expression of immune-related genes in the spleen, which significantly protected the liver, brain and spleen from pathological damage. In addition, PDH antiserum could significantly inhibit the growth of S. suis and the formation of S. suis biofilm in vitro. These results further suggest that PDH is a promising candidate for S. suis biofilm-related subunit vaccine.

The otc gene of Streptococcus suis plays an important role in biofilm formation, adhesion, and virulence in a murine model.

Streptococcus suis (S. suis) is an emerging zoonotic pathogen that can cause meningitis, arthritis, pneumonia, and sepsis. It poses a serious threat to the swine industry and public health worldwide. Ornithine carbamoyltransferase (OTC) is involved in the arginine deiminase system. OTC, which is a widely distributed enzyme in microorganisms, mammals, and higher plants, catalyzes the conversion of ornithine to citrulline. The present study showed that the otc gene plays an important role in the pathogenesis of S. suis infections. The ability of an otc-deficient mutant (Δotc) to form a biofilm was significantly reduced compared to the wild-type (WT) strain, as determined by crystal violet staining. Confocal laser scanning microscopy and scanning electron microscopy observations showed that the weakening of biofilm formation by the Δotc strain is related to a decrease in the extracellular matrix. In addition, compared to the WT strain, the Δotc strain had a reduced capacity to adhere to human laryngeal epidermoid carcinoma (HEp-2) cells compared to the WT strain. A real-time PCR analysis showed that the expression of adhesion-related genes by the Δotc strain was also lower than that of the WT strain. The virulence of the Δotc strain was significantly lower than that of the WT strain in a murine infection model. In addition, a histological analysis showed that the pathogenicity of the Δotc strain was lower than that of the WT strain, causing only slight inflammatory lesions in lung, liver, spleen, and kidney tissues. No significant differences were observed between the complemented mutant (CΔotc) and WT strains with respect to biofilm formation, adhesion, gene expression, and virulence. The present study provided evidence that the otc gene plays a pivotal role in the regulation of S. suis adhesion and biofilm formation. It also suggested that the otc gene is indirectly involved in the pathogenesis of S. suis serotype 2 infections.

Autoinducer-2 influences tetracycline resistance in Streptococcus suis by regulating the tet(M) gene via transposon Tn916.

The concern over increasing resistance to tetracyclines (TCs), such as tetracycline and chlortetracycline, necessitates exploration of new approaches to combating infection in antimicrobial therapy. Given that bacteria use the chemical language of autoinducer 2 (AI-2) signaling molecules in order to communicate and regulate group behaviors, we asked whether the AI-2 signaling influence the tetracyclines antibiotics susceptibility in S. suis. Our present work demonstrated that MIC increased when exogenous AI-2 was added, when compared to the wild type strain. When grown in the presence of sub-MIC of antibiotics, it has been shown that exogenous AI-2 increases growth rate and biofilm formation. These results suggest that the TCs resistance in S. suis could involve a signaling mechanism. Base on the above observations, transcriptomic analyses showed significant differences in the expression of tet(M) of tetracyclines resistance genes, as well as differences in Tn916 transposon related genes transcription, as judged by RT-PCR. Our results provide strong evidence that AI-2 signaling molecules is may involve in TCs antibiotic resistance in S. suis by regulating tet(M) gene via Tn916 transposon. This study may suggest that targeting AI-2 signaling in bacteria could represent an alternative approach in antimicrobial therapy.

LuxS/AI-2 system is involved in fluoroquinolones susceptibility in Streptococcus suis through overexpression of efflux pump SatAB.

Increasing resistance to fluoroquinolones (FQs), such as norfloxacin and enrofloxacin, supports the need for the discovery of novel molecules and alternative approaches in antimicrobial therapy. Quorum sensing (QS) is a promising target for next-generation anti-infective agents designed to address the evolving drug resistance in bacterial pathogens. Given that the LuxS/autoinducer-2 (AI-2) quorum-sensing system regulates microbial group behaviors, we hypothesized that this system influences the FQ susceptibility in Streptococcus suis. It was found that a luxS mutant (ΔluxS) of S. suis possesses an increased susceptibility to FQs compared to the wild type strain. When grown in the presence of sub-MIC of antibiotics, the ΔluxS strain showed a significant decrease in growth rate and biofilm formation. These results suggest that the FQ resistance in S. suis could involve a signaling mechanism associated with the LuxS/AI-2 quorum-sensing system. HPLC (High Performance Liquid Chromatography) analyses showed a significant increase in the intracellular accumulation of enrofloxacin in the ΔluxS strain compared to the wild type strain. This increase was less pronounced in the presence of exogenous AI-2. Moreover, the expression of satA and satB genes was decreased in the ΔluxS strain. Exogenous AI-2 reversed the down-regulated gene expression observed in the ΔluxS strain. Our study brought strong evidence that the LuxS/AI-2 system in S. suis is involved in FQ susceptibility by regulating the efflux pump SatAB. LuxS is highly conserved among Gram-positive bacteria and may therefore represent a novel antimicrobial target for an alternative approach in antimicrobial therapy.