α-Enolase of Streptococcus pneumoniae Induces Formation of Neutrophil Extracellular Traps

Mori, Yoshiyuki; Yamaguchi, M.; Terao, Yutaka; Hamada, Shigeyuki; Ooshima, Takashi; Kawabata, Shigetada

doi:10.1074/jbc.m111.280321

Cited by 102 publications

(82 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…1, A and B). In addition to its previously reported functions, our findings suggest that PepO is multifunctional in both cytoplasm and the extracellular milieu, similar to the properties of GAPDH and enolase, which lack a signal sequence but are found in culture supernatant (27)(28)(29)(30)(31). The present study provides clues about the variable ability of S. pyogenes strains to express and secrete PepO.…”

Section: Discussionmentioning

confidence: 67%

Streptococcus pyogenes Endopeptidase O Contributes to Evasion from Complement-mediated Bacteriolysis via Binding to Human Complement Factor C1q

Honda-Ogawa

Sumitomo

Mori

et al. 2017

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Edited by Luke O'NeillStreptococcus pyogenes secretes various virulence factors for evasion from complement-mediated bacteriolysis. However, full understanding of the molecules possessed by this organism that interact with complement C1q, an initiator of the classical complement pathway, remains elusive. In this study, we identified an endopeptidase of S. pyogenes, PepO, as an interacting molecule, and investigated its effects on complement immunity and pathogenesis. Enzyme-linked immunosorbent assay and surface plasmon resonance analysis findings revealed that S. pyogenes recombinant PepO bound to human C1q in a concentration-dependent manner under physiological conditions. Sites of inflammation are known to have decreased pH levels, thus the effects of PepO on bacterial evasion from complement immunity was analyzed in a low pH condition. Notably, under low pH conditions, PepO exhibited a higher affinity for C1q as compared with IgG, and PepO inhibited the binding of IgG to C1q. In addition, pepO deletion rendered S. pyogenes more susceptible to the bacteriocidal activity of human serum. Also, observations of the morphological features of the pepO mutant strain (⌬pepO) showed damaged irregular surfaces as compared with the wild-type strain (WT). WT-infected tissues exhibited greater severity and lower complement activity as compared with those infected by ⌬pepO in a mouse skin infection model. Furthermore, WT infection resulted in a larger accumulation of C1q than that with ⌬pepO. Our results suggest that interaction of S. pyogenes PepO with C1q interferes with the complement pathway, which enables S. pyogenes to evade complement-mediated bacteriolysis under acidic conditions, such as seen in inflammatory sites.The human pathogen Streptococcus pyogenes is a ␤-hemolytic Gram-positive bacterium that causes a wide variety of diseases in various anatomical sites. Superficial infection of S. pyogenes generally leads to local suppurative lesions, such as seen in pharyngitis and impetigo cases. Such mucosal and skin infections are occasionally followed by onset of immune sequelae, including acute rheumatic fever and acute glomerulonephritis. Furthermore, S. pyogenes infection can also cause invasive diseases, such as necrotizing fasciitis, cellulitis, bacteremia, and streptococcal toxic shock syndrome (STSS), 2 with a high mortality rate, which has been a serious problem throughout the world (1-3).Host possesses various immune systems such as the complement system, one of the most important components of innate immunity, that contributes to host defense against pathogens, especially in the early stage of infection (4). Recognition of an invading pathogen is the trigger for complement pathway activation, and then each complement factor is activated in a sequential manner with precise control (5). Together with opsonization, complement-mediated direct killing of pathogens is attributable to formation of the membrane attack complex (MAC), which consists of late complement factors and induces bacteriolysis by pore formati...

show abstract

Section: Discussionmentioning

confidence: 67%

Streptococcus pyogenes Endopeptidase O Contributes to Evasion from Complement-mediated Bacteriolysis via Binding to Human Complement Factor C1q

Honda-Ogawa

Sumitomo

Mori

et al. 2017

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…Although unproven, the events involved in induction of NETosis in this setting are likely to be related to the pro-NETotic actions of Ply described in the current study, augmented possibly by pneumococcal alpha-enolase, which has also been reported to induce NET formation, albeit via an indirect mechanism [41]. Although NETs are intended to trap and possibly kill microbial pathogens, the pneumococcus via expression of the polysaccharide capsule [42] and the surface endonuclease, EndA [43], is particularly adept at evading and escaping from NETs.…”

Section: Discussionmentioning

confidence: 97%

Pneumolysin activates neutrophil extracellular trap formation

Nel¹,

Theron

Durandt

et al. 2016

Clinical and Experimental Immunology

View full text Add to dashboard Cite

SummaryThe primary objective of the current study was to investigate the potential of the pneumococcal toxin, pneumolysin (Ply), to activate neutrophil extracellular trap (NET) formation in vitro. Isolated human blood neutrophils were exposed to recombinant Ply (5-20 ng ml 21) for 30-90 min at 378C and NET formation measured using the following procedures to detect extracellular DNA: (i) flow cytometry using VybrantV R DyeCycle TM Ruby; (ii) spectrofluorimetry using the fluorophore, Sytox V R Orange (5 lM); and (iii) NanoDrop V R technology. These procedures were complemented by fluorescence microscopy using 4 0 , 6-diamino-2-phenylindole (DAPI) (nuclear stain) in combination with anti-citrullinated histone monoclonal antibodies to visualize nets. Exposure of neutrophils to Ply resulted in relatively rapid (detected within 30-60 min), statistically significant (P < 0Á05) dose-and time-related increases in the release of cellular DNA impregnated with both citrullinated histone and myeloperoxidase. Microscopy revealed that NETosis appeared to be restricted to a subpopulation of neutrophils, the numbers of NET-forming cells in the control and Ply-treated systems (10 and 20 ng ml 21 ) were 4Á3 (4Á2), 14.3 (9Á9) and 16Á5 (7Á5), respectively (n 5 4, P < 0Á0001 for comparison of the control with both Ply-treated systems). Ply-induced NETosis occurred in the setting of retention of cell viability, and apparent lack of involvement of reactive oxygen species and Toll-like receptor 4. In conclusion, Ply induces vital NETosis in human neutrophils, a process which may either contribute to host defence or worsen disease severity, depending on the intensity of the inflammatory response during pneumococcal infection.

show abstract

“…An S. pneumoniae ccs4 (locus_tag: SP_0200, GenBank: AAK74380.1) mutant strain (Δ ccs4 ) was constructed as previously described [49,50]. Briefly, the upstream region of ccs4 , an aad9 cassette, and the downstream region of ccs4 were combined by PCR, then the combined product was used to obtain mutant strains.…”

Section: Methodsmentioning

confidence: 99%

Competence-induced protein Ccs4 facilitates pneumococcal invasion into brain tissue and virulence in meningitis

et al. 2018

Self Cite

View full text Add to dashboard Cite

Streptococcus pneumoniae is a major pathogen that causes pneumonia, sepsis, and meningitis. The candidate combox site 4 (ccs4) gene has been reported to be a pneumococcal competence-induced gene. Such genes are involved in development of S. pneumoniae competence and virulence, though the functions of ccs4 remain unknown. In the present study, the role of Ccs4 in the pathogenesis of pneumococcal meningitis was examined. We initially constructed a ccs4 deletion mutant and complement strains, then examined their association with and invasion into human brain microvascular endothelial cells. Wild-type and Ccs4-complemented strains exhibited significantly higher rates of association and invasion as compared to the ccs4 mutant strain. Deletion of ccs4 did not change bacterial growth activity or expression of NanA and CbpA, known brain endothelial pneumococcal adhesins. Next, mice were infected either intravenously or intranasally with pneumococcal strains. In the intranasal infection model, survival rates were comparable between wild-type strain-infected and ccs4 mutant strain-infected mice, while the ccs4 mutant strain exhibited a lower level of virulence in the intravenous infection model. In addition, at 24 hours after intravenous infection, the bacterial burden in blood was comparable between the wild-type and ccs4 mutant strain-infected mice, whereas the wild-type strain-infected mice showed a significantly higher bacterial burden in the brain. These results suggest that Ccs4 contributes to pneumococcal invasion of host brain tissues and functions as a virulence factor.

show abstract

α-Enolase of Streptococcus pneumoniae Induces Formation of Neutrophil Extracellular Traps

Cited by 102 publications

References 36 publications

Streptococcus pyogenes Endopeptidase O Contributes to Evasion from Complement-mediated Bacteriolysis via Binding to Human Complement Factor C1q

Streptococcus pyogenes Endopeptidase O Contributes to Evasion from Complement-mediated Bacteriolysis via Binding to Human Complement Factor C1q

Pneumolysin activates neutrophil extracellular trap formation

Competence-induced protein Ccs4 facilitates pneumococcal invasion into brain tissue and virulence in meningitis

Contact Info

Product

Resources

About