Related Experiment Videos
Determination of alternate pathway complement kinetics by electron spin resonance spectroscopy
American Journal of Clinical Pathology
|June 1, 1983
Summary
This study introduces electron spin resonance (ESR) spin trapping to measure neutrophil metabolic bursts. The technique reveals insights into the alternate complement pathway kinetics, unaffected by C8 complement protein absence.
Area of Science:
- Immunology
- Biochemistry
- Analytical Chemistry
Background:
- Neutrophil metabolic burst is crucial for immune response.
- Electron spin resonance (ESR) spin trapping offers a method to detect reactive oxygen species.
- Understanding complement cascade kinetics is vital for immunology.
Purpose of the Study:
- To develop a novel technique for measuring neutrophil metabolic burst kinetics using ESR spin trapping.
- To establish a functional assay for the kinetics of the alternate pathway complement cascade.
- To investigate the role of complement C8 in opsonization kinetics.
Main Methods:
- Peripheral blood neutrophils (PMN) were stimulated with opsonized zymosan (OpZym).
- Free hydroxyl radicals generated during the metabolic burst were trapped using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO).
- A computer-controlled ESR spectrometer was used for rapid, sequential measurements of DMPO-hydroxyl radical adducts.
Main Results:
- The ESR spin trapping technique successfully measured the kinetics of the neutrophil metabolic burst.
- Differences in spin adduct formation with OpZym versus unopsonized zymosan (Zym) provided opsonization kinetics information.
- Alternate complement pathway opsonization kinetics in normal and C8-deficient sera showed no significant difference due to C8 absence.
Conclusions:
- ESR spin trapping is a viable method for assessing neutrophil metabolic burst kinetics.
- The developed assay effectively characterizes alternate complement pathway opsonization kinetics.
- Complement C8 protein does not significantly impact the kinetics of alternate pathway opsonization.