Factor affecting the in vitro assessment of opsonization: a study of the kinetics of opsonization using the technique of phagocytic chemiluminescence.
A J Williams, M. Hastings, CHARLES S. F. EASMON, PETER J. COLE
Abstract
A J Williams, M. Hastings, CHARLES S. F. EASMON, PETER J. COLE
Abstract
The importance of several factors involved in the investigation of opsonic defects was studied using phagocytic Luminol-dependent chemiluminescence. The range for the opsonization of zymosan and bakers' yeast by serum from healthy individuals was wide and kinetic studies showed comparative differences for different periods of incubation, serum concentrations and particles. Decay in the opsonic activity of serum stored at different temperatures was demonstrated and its clinical implications emphasized. By using techniques to ablate independently the classical and alternative pathways of complement activation, the contribution of these to the opsonization of zymosan, Staphylococcus aureus (NCTC 6571), Pseudomonas aeruginosa and group B streptococcus (NCTC 11080) by normal and hypogammaglobulinaemic serum at a concentration of 7% was assessed. By comparison of the results obtained for different periods of incubation between particle and serum, the need for consideration of this parameter when assessing opsonic activity was shown. The results using the chemiluminescence assay were compared with those using other methods and were found to correlate well.
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The importance of several factors involved in the investigation of opsonic defects was studied using phagocytic Luminol-dependent chemiluminescence. The range for the opsonization of zymosan and bakers' yeast by serum from healthy individuals was wide and kinetic studies showed comparative differences for different periods of incubation, serum concentrations and particles. Decay in the opsonic activity of serum stored at different temperatures was demonstrated and its clinical implications emphasized. By using techniques to ablate independently the classical and alternative pathways of complement activation, the contribution of these to the opsonization of zymosan, Staphylococcus aureus (NCTC 6571), Pseudomonas aeruginosa and group B streptococcus (NCTC 11080) by normal and hypogammaglobulinaemic serum at a concentration of 7% was assessed. By comparison of the results obtained for different periods of incubation between particle and serum, the need for consideration of this parameter when assessing opsonic activity was shown. The results using the chemiluminescence assay were compared with those using other methods and were found to correlate well.
Key concepts: Antibody opsonization, Opsonin, Zymosan, Chemiluminescence, Phagocytosis, Incubation, Microbiology, Pseudomonas aeruginosa