Histopathologic changes in kidney and liver correlate with streptococcal pyrogenic exotoxin B production in the mouse model of group A streptococcal infection

Kuo, Chih Feng; Luo, Yueh Hsia; Lin, Hsiu Yueh; Huang, Kao-Jean; Wu, Jiunn Jong; Lei, Huan Yao; Lin, Ming Tsan; Chuang, Woei Jer; Liu, Ching Chuan; Jin, Ying; Lin, Yee Shin

doi:10.1016/j.micpath.2004.01.003

Cited by 60 publications

(91 citation statements)

References 52 publications

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“…Immune sera were collected 7 days after the second boost and used for subsequent experiments. The sera anti-MF3 titers were measured by enzyme-linked immunosorbent assay as described previously (23).…”

Section: Bacterial Strains Growth Conditions and Sample Preparation-mentioning

confidence: 99%

“…All RNA samples were reverse transcribed for 30 min at 42°C with a High Capacity cDNA Reverse Transcription Kit according to the standard protocol of the supplier (Applied Biosystems, Foster City, CA). Real-time RT-PCR was performed on an AB PRISM 7300 Sequence Detection System (Applied Biosystems) using the QuantiTect SYBR green RT-PCR kit (Qiagen) as described (23). Expression levels of each test gene were normalized to those of 16 S rRNA, which did not show any change in expression as a consequence of perR mutation.…”

Section: Bacterial Strains Growth Conditions and Sample Preparation-mentioning

confidence: 99%

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Differential Secretomics of Streptococcus pyogenes Reveals a Novel Peroxide Regulator (PerR)-regulated Extracellular Virulence Factor Mitogen Factor3 (MF3)

Wen

Tsou

Kuo

et al. 2011

Molecular & Cellular Proteomics

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Streptococcus pyogenes is a human pathogen that causes various diseases. Numerous virulence factors secreted by S. pyogenes are involved in pathogenesis. The peroxide regulator (PerR) is associated with the peroxide resistance response and pathogenesis, but little is known about the regulation of the secretome involved in virulence. To investigate how PerR regulates the expression of the S. pyogenes secretome involved in virulence, a perR deficient mutant was used for comparative secretomic analysis with a wild-type strain. The conditioned medium containing secreted proteins of a wild-type strain and a perR deficient mutant at the stationary phase were collected for two-dimensional gel electrophoresis analysis, where protease inhibitors were applied to avoid the degradation of extracellular proteins. Differentially expressed protein spots were identified by liquid chromatography electrospray ionization tandem MS. More than 330 protein spots were detected on each gel. We identified 25 unique up-regulated proteins and 13 unique downregulated proteins that were directly or indirectly controlled by the PerR regulator. Among these identified proteins, mitogen factor 3 (MF3), was selected to verify virulence and the expression of gene products. The data showed that MF3 protein levels in conditioned medium, as measured by immunoblot analysis, correlated well with protein levels determined by two-dimensional gel electrophoresis analysis. We also demonstrated that PerR bound to the promoter region of the mf3 gene. The result of an infection model showed that virulence was attenuated in the mf3 deficient mutant. Additional growth data of the wild-type strain and the mf3 deficient mutant suggested that MF3 played a role in digestion of exogenous DNA for promoting growth. To summarize, we conclude that PerR can positively regulate the expression of the secreted protein MF3 that contributes to the virulence in S. pyogenes. The analysis of the PerR-regulated secretome provided key information for the elucidation of the host-pathogen interactions and might assist in the

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Section: Bacterial Strains Growth Conditions and Sample Preparation-mentioning

confidence: 99%

Section: Bacterial Strains Growth Conditions and Sample Preparation-mentioning

confidence: 99%

Differential Secretomics of Streptococcus pyogenes Reveals a Novel Peroxide Regulator (PerR)-regulated Extracellular Virulence Factor Mitogen Factor3 (MF3)

Wen

Tsou

Kuo

et al. 2011

Molecular & Cellular Proteomics

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“…Two micrograms of total protein was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by transfer to polyvinylidene difluoride membranes. The membrane was blocked with phosphate-buffered saline (PBS) containing 5% (wt/vol) skim milk at 37°C for 2 h and probed with anti-C192S mouse serum prepared as previously described (28). After three washes with PBS containing 0.05% Tween 20 (PBST), the membrane was incubated with horseradish peroxidase-conjugated anti-mouse immunoglobulin antibody (Chemicon International, Inc., Temecula, CA).…”

Section: Methodsmentioning

confidence: 99%

emm1/Sequence Type 28 Strains of Group A Streptococci That ExpresscovRat Early Stationary Phase Are Associated with Increased Growth and Earlier SpeB Secretion

Chiang-Ni

Zheng²,

et al. 2009

J Clin Microbiol

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Streptococcus pyogenes (group A streptococcus [GAS]) is a versatile human pathogen, and emm1/sequence type 28 (ST28) is the most frequently isolated type from GAS infections. The emm1/ST28 strain is associated with necrotizing fasciitis and streptococcal toxic shock syndrome. Growth-phase regulation is one of the important regulatory mechanisms in GAS, which controls gene expression at restricted phases of growth. CovRS, a two-component regulatory system, is considered the regulator of streptococcal pyrogenic exotoxin B (SpeB) and is thought to be activated in the exponential phase of growth. In the present study, Northern hybridization analysis showed that 52% of the analyzed GAS strains expressed covR at the exponential phase, but 48% of the strains expressed covR at the early stationary phase of growth. Strains transcribing covR at the early stationary phase showed better growth and earlier SpeB expression than the other group of strains. Multilocus sequence typing and pulsed-field gel electrophoresis analysis showed only emm1/ST28 strains (which comprise a clonal cluster) were expressing covR at the early stationary phase of growth, indicating that emm1/ST28 strains have special characteristics which may be related to their worldwide distribution.

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“…SPE B digests a number of host proteins, including immunoglobulin (8,10,14,45), kininogen (17), matrix metalloprotease (3), urokinase receptor (51), fibrinogen (33), and fibronectin as well as vitronectin (25) and interleukin-1 precursor (24). A speB mutant strain has decreased resistance to neutrophil phagocytosis, dissemination to organs, and mortality in the mouse model (26,28,30,31,32). The exact mechanism by which SPE B inhibits neutrophil phagocytosis has yet to be clearly defined.…”

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confidence: 99%

Degradation of Complement 3 by Streptococcal Pyrogenic Exotoxin B Inhibits Complement Activation and Neutrophil Opsonophagocytosis

Kuo¹,

Lin²,

Chuang³

et al. 2008

Infect Immun

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Streptococcal pyrogenic exotoxin B (SPE B), a cysteine protease, is an important virulence factor in group A streptococcus (GAS) infection. The inhibition of phagocytic activity by SPE B may help prevent bacteria from being ingested. In this study, we examined the mechanism SPE B uses to enable bacteria to resist opsonophagocytosis. Using an enzyme-linked immunosorbent assay, we found that SPE B-treated serum impaired the activation of the classical, the lectin, and the alternative complement pathways. In contrast, C192S, a SPE B mutant lacking protease activity, had no effect on complement activation. Further study showed that cleavage of serum C3 by SPE B, but not C192S, blocked zymosan-induced production of reactive oxygen species in neutrophils as a result of decreased deposition of C3 fragments on the zymosan surface. Reconstitution of C3 into SPE B-treated serum unblocked zymosan-mediated neutrophil activation dose dependently. SPE Btreated, but not C192S-treated, serum also impaired opsonization of C3 fragments on the surface of GAS strain A20. Moreover, the amount of C3 fragments on the A20 cell surface, a SPE B-producing strain, was less than that on its isogenic mutant strain, SW507, after opsonization with normal serum. A20 opsonized with SPE B-treated serum was more resistant to neutrophil killing than A20 opsonized with normal serum, and SPE B-mediated resistance was C3 dependent. These results suggest a novel SPE B mechanism, one which degrades serum C3 and enables GAS to resist complement damage and opsonophagocytosis.

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Histopathologic changes in kidney and liver correlate with streptococcal pyrogenic exotoxin B production in the mouse model of group A streptococcal infection

Cited by 60 publications

References 52 publications

Differential Secretomics of Streptococcus pyogenes Reveals a Novel Peroxide Regulator (PerR)-regulated Extracellular Virulence Factor Mitogen Factor3 (MF3)

Differential Secretomics of Streptococcus pyogenes Reveals a Novel Peroxide Regulator (PerR)-regulated Extracellular Virulence Factor Mitogen Factor3 (MF3)

emm1/Sequence Type 28 Strains of Group A Streptococci That ExpresscovRat Early Stationary Phase Are Associated with Increased Growth and Earlier SpeB Secretion

Degradation of Complement 3 by Streptococcal Pyrogenic Exotoxin B Inhibits Complement Activation and Neutrophil Opsonophagocytosis

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