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Updated: Aug 4, 2026

Measuring the 50% Haemolytic Complement (CH50) Activity of Serum
Published on: March 29, 2010
Similarities between complement-mediated and streptolysin S-mediated hemolysis
A Carr1, D D Sledjeski, A Podbielski
1Department of Microbiology & Immunology, Medical College of Ohio, Toledo, Ohio 43614-5806, USA.
Abstract:
The oxygen-stable hemolysin streptolysin S (SLS) of Streptococcus pyogenes is encoded in part by the pel/sagA gene product. Antibodies to a synthetic peptide from the C terminus of the Pel/SagA open reading frame inhibited hemolysis mediated by both culture supernatants from multiple M serotypes of S. pyogenes isolates or a commercially available SLS preparation. Analysis of the SLS-mediated hemolytic reaction demonstrated that it was temperature- and concentration-dependent. Like complement-mediated hemolysis it conforms to the prediction of a one-hit mechanism of hemolysis. A number of intermediates in the SLS-mediated hemolysis of sheep erythrocytes could be distinguished. SLS could bind to erythrocytes below 17 degrees C; however, lysis could only occur at temperatures >23 degrees C. Following binding of SLS and washing, a papain-sensitive intermediate could be distinguished prior to insertion of the SLS complex into the erythrocyte membrane, which resulted in formation of a transmembrane pore and led to irreversible osmotic lysis of the cell. These intermediates were similar to those described previously during complement-mediated hemolysis.
Insights
Streptococcus pyogenes streptolysin S (SLS) hemolysin activity depends on temperature and concentration. Its mechanism involves distinct intermediates, similar to complement-mediated hemolysis, leading to erythrocyte lysis.
Area of Science:
- Microbiology
- Immunology
- Biochemistry
Background:
- Streptococcus pyogenes produces streptolysin S (SLS), an oxygen-stable hemolysin.
- SLS is implicated in the pathogenesis of S. pyogenes infections.
- The pel/sagA gene product is involved in encoding SLS.
Purpose of the Study:
- To investigate the mechanism of streptolysin S (SLS)-mediated hemolysis.
- To characterize the temperature and concentration dependence of SLS activity.
- To identify and compare intermediates in the SLS hemolytic pathway with complement-mediated hemolysis.
Main Methods:
- Antibody inhibition assays using synthetic peptides from Pel/SagA.
- Analysis of hemolytic reactions under varying temperature and concentration conditions.
- Distinguishing intermediates in the erythrocyte lysis process.
Main Results:
- Antibodies against a C-terminal peptide of Pel/SagA inhibited SLS hemolysis.
- SLS-mediated hemolysis is temperature- and concentration-dependent, following a one-hit mechanism.
- Distinct intermediates were identified, including temperature-dependent binding and papain-sensitive stages, prior to pore formation and lysis.
Conclusions:
- SLS-mediated hemolysis involves specific binding and intermediate stages before irreversible cell lysis.
- The identified intermediates share similarities with those observed in complement-mediated hemolysis.
- Understanding these mechanisms provides insights into S. pyogenes pathogenesis and potential therapeutic targets.
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