Proinflammatory cytokine and chemokine modulation by Streptococcus suis in a whole-blood culture system.
ABSTRACT Streptococcus suis is an important swine and human pathogen. Inflammation, a hallmark of S. suis infection, is thought to be responsible for most clinical signs of meningitis, septicaemia and sudden death. In this work, using a porcine whole blood model, S. suis serotype 2 was shown to trigger the release of several pro-inflammatory cytokines as evaluated by reverse transcriptase-PCR and enzyme-linked immunosorbent assay. Although individual variations were observed among different S. suis strains, no correlations were observed between the strain origin/phenotype and cytokine levels. Live bacteria induced higher tumour necrosis factor alpha, interleukin-1 beta (IL-1beta) and IL-6 levels than did heat-killed bacteria. In contrast, heat-killed bacteria stimulated higher levels of IL-8 and monocyte chemotactic protein one (MCP-1). The bacterial cell wall was observed to be the major cytokine-inducting components, whereas capsule expression was important for MCP-1 activation. The presence of specific antibodies suppressed bacterial growth resulting in significantly reduced levels of cytokine production. Thus, antibody-mediated bacterial phagocytosis combined with suppressed inflammation may be beneficial for infection control strategies. We provide first evidence of S.suis-induction of pro-inflammatory swine cytokines and demonstrate the strength and relevance of the whole blood culture systems in the investigation of S. suis modulation of cytokine production.
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ABSTRACT: Streptococcus suis capsular type 2 is an important zoonotic agent of meningitis. Previous studies reported that, in contrast to nonencapsulated mutants, encapsulated S. suis is able to resist phagocytosis. However, the mechanisms by which S. suis avoids phagocytosis are unknown. To elucidate the signaling pathway(s) involved in S. suis antiphagocytosis, we compared the ability of an encapsulated strain and its nonencapsulated mutant to induce the activation of Akt and protein kinase C (PKC), which are downstream kinases of the phosphatidylinositol 3-kinase (PI-3K) pathway, known to be involved in the phagocytosis processes. The results demonstrated high levels of Akt and PKCalpha phosphorylation after infection of J774 macrophages with the nonencapsulated mutant, whereas the encapsulated strain showed reduced activation of PI-3K/Akt/PKCalpha signaling pathway, as well as several protein tyrosine events. These results correlated with the number of intracellular bacteria. Macrophages pretreated with specific PI-3K or PKC inhibitors showed reduced levels of Akt and PKCalpha phosphorylation, resulting in 50% reduction of phagocytosis. The role of phosphatases in the antiphagocytic mechanisms was evaluated by using phosphatase inhibitors, as well as SHP-1-deficient macrophages. Only in the absence of SHP-1 did the phagocytosis of encapsulated S. suis significantly increase, leading to Akt phosphorylation levels similar to those observed with the nonencapsulated strain, indicating activation of this important SH2 domain-containing tyrosine phosphatase by encapsulated S. suis. Finally, when purified S. suis capsular polysaccharide (CPS) was added to macrophages, no phosphorylation events were observed. In addition, CPS and encapsulated S. suis were able to inhibit the uptake of the nonencapsulated mutant. These results suggest the importance of CPS in the mechanisms, whereby S. suis downmodulates phagocytosis.Infection and Immunity 10/2004; 72(9):5322-30. · 4.07 Impact Factor
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ABSTRACT: The genetic diversity of 88 Streptococcus suis serotype 2 isolates which were recovered from various countries was examined by randomly amplified polymorphic DNA (RAPD) analysis with three primers. This bacterial collection included 80 isolates of porcine origin and 8 of human origin. This investigation allowed the identification of 23 RAPD types containing 1 to 30 isolates originating from one to six countries. Common RAPD patterns were found between human and pig isolates. The isolates were also tested for the production of virulent factors such as hemolysin, muramidase-released protein (MRP), and extracellular factor (EF). All isolates exhibiting the virulent phenotype hemolysin+ MRP+ EF+ clearly clustered on the basis of fingerprinting by RAPD analysis. In a similar way, most of isolates with the hemolysin- MRP- EF- phenotype were assigned to one RAPD cluster. Therefore, RAPD clusters are more related to the phenotype defined with hemolysin, MRP, and EF than to the geographic origin of the isolates. These data indicate that RAPD analysis used in conjunction with phenotypic methods provides a reliable method for the assessment of the clonal relationship between S. suis isolates responsible for infections in pigs or humans, especially for those exhibiting the classic "virulent" phenotype hemolysin+ MRP+ EF+.Journal of Clinical Microbiology 03/1999; 37(2):362-6. · 4.07 Impact Factor
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ABSTRACT: A defined allelic-replacement mutant of the sly gene, encoding a thiol-activated cytolysin, from a European isolate of Streptococcus suis serotype 2 was generated and characterized. Unlike the parental strain, it is nonhemolytic, noncytotoxic for cultured macrophage-like cells, avirulent in a mouse infection model, yet only slightly attenuated in a porcine model of systemic infection.Infection and Immunity 05/2001; · 4.07 Impact Factor