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1.
J Biol Chem ; 283(10): 6253-60, 2008 Mar 07.
Article in English | MEDLINE | ID: mdl-18160402

ABSTRACT

A relative lack of neutrophils around Streptococcus pyogenes is observed in streptococcal toxic shock syndrome (STSS). Because the bacteria spread rapidly into various organs in STSS, we speculated that S. pyogenes is equipped with molecules to evade the host innate immune system. Complement C3b opsonizes the pathogen to facilitate phagocytosis, and a complex of C3b converts C5 into anaphylatoxin. Because we found that C3 (C3b) is degraded in sera from patients with STSS, we investigated the mechanism of C3 (C3b) degradation by S. pyogenes. We incubated human C3b or serum with recombinant SpeB (rSpeB), a wild-type S. pyogenes strain isolated from an STSS patient or its isogenic DeltaspeB mutant and examined the supernatant by Western blotting with anti-human C3b. Western blot and Biacore analyses revealed that rSpeB and wild-type S. pyogenes rapidly degrade C3b. Additionally, C3 (C3b) was not detected in sera collected from infected areas of STSS patients. Furthermore, the survival rate in human blood and in mice was lower for the DeltaspeB mutant than the wild-type strain. Histopathological observations demonstrated that neutrophils were recruited to and phagocytosed the DeltaspeB mutant, whereas with the wild-type strain, few neutrophils migrated to the site of infection, and the bacteria spread along the fascia. We observed the degradation of C3 (C3b) in sera from STSS patients and the degradation of C3 (C3b) by rSpeB. This suggests that SpeB contributes to the escape of S. pyogenes from phagocytosis at the site of initial infection, allowing it to invade host tissues during severe infections.


Subject(s)
Bacterial Proteins/immunology , Complement C3b/immunology , Exotoxins/immunology , Neutrophils/immunology , Phagocytosis/immunology , Shock, Septic/immunology , Streptococcus pyogenes/immunology , Adult , Anaphylatoxins/analysis , Anaphylatoxins/immunology , Animals , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Complement C3b/analysis , Complement C5/analysis , Complement C5/immunology , Exotoxins/genetics , Exotoxins/metabolism , Female , Gene Deletion , Humans , Male , Mice , Middle Aged , Neutrophils/metabolism , Phagocytosis/genetics , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/metabolism , Shock, Septic/blood
3.
Infect Immun ; 70(6): 3227-33, 2002 Jun.
Article in English | MEDLINE | ID: mdl-12011018

ABSTRACT

In Japan, more than 10% of streptococcal toxic shock-like syndrome (TSLS) cases have been caused by Streptococcus pyogenes M3/T3 isolates since the first reported TSLS case in 1992. Most M3/T3 isolates from TSLS or severe invasive infection cases during 1992 to 2001 and those from noninvasive cases during this period are indistinguishable in pulsed-field gel electropherograms. The longest fragments of these recent isolates were 300 kb in size, whereas those of isolates recovered during or before 1973 were 260 kb in size. These 260- and 300-kb fragments hybridized to each other, suggesting the acquisition of an about 40-kb fragment by the recent isolates. The whole part of the acquired fragment was cloned from the first Japanese TSLS isolate, NIH1, and its nucleotide sequence was determined. The 41,796-bp fragment is temperate phage phiNIH1.1, containing a new superantigen gene speL near its right attachment site. The C-terminal part of the deduced amino acid sequence of speL has 48 and 46% similarity with well-characterized erythrogenic toxin SpeC and the most potent superantigen, SmeZ-2, respectively. None of 10 T3 isolates recovered during or before 1973 has speL, whereas all of 18 M3/T3 isolates recovered during or after 1992 and, surprisingly, Streptococcus equi subsp. equi ATCC 9527 do have this gene. Though plaques could not be obtained from phiNIH1.1, its DNA became detectable from the phage particle fraction upon mitomycin C induction, showing that this phage is not defective. A horizontal transfer of the phage carrying speL may explain the observed change in M3/T3 S. pyogenes isolates in Japan.


Subject(s)
Antigens, Bacterial/genetics , Bacterial Proteins , Shock, Septic/microbiology , Streptococcus Phages/genetics , Streptococcus pyogenes/genetics , Superantigens/genetics , Amino Acid Sequence , Base Sequence , Cloning, Molecular , DNA, Bacterial , Electrophoresis, Gel, Pulsed-Field/methods , Humans , Japan/epidemiology , Molecular Sequence Data , Sequence Homology, Amino Acid , Shock, Septic/epidemiology , Streptococcus pyogenes/isolation & purification , Streptococcus pyogenes/pathogenicity , Streptococcus pyogenes/virology
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