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Phagocytic cell chemiluminescence using different zymosan preparations
D S Gridley1, B H Lau, J M Tosk
1Department of Microbiology, Loma Linda University School of Medicine, California 92350.
Journal of Clinical Laboratory Analysis
|January 1, 1991
Summary
Laboratory-prepared zymosans effectively induce chemiluminescence (CL) in phagocytic cells. These zymosans, whether opsonized or not, show superior or equal performance compared to commercial preparations in stimulating oxidative burst responses.
Area of Science:
- Immunology
- Cell Biology
Background:
- Phagocytic cells play a crucial role in innate immunity.
- Chemiluminescence (CL) assays are used to measure oxidative burst activity in phagocytes.
- Zymosan is a common particulate activator used in CL assays.
Purpose of the Study:
- To compare the efficacy of laboratory-prepared opsonized and unopsonized zymosan with a commercial preparation.
- To evaluate the induction of luminol-dependent oxidative burst in various phagocytic cell populations.
Main Methods:
- Preparation of opsonized and unopsonized zymosan in the laboratory.
- Testing CL production using an automated luminometer with human whole blood, isolated human neutrophils, mouse splenocytes, and a macrophage cell line (J774A.1).
- Utilizing recombinant murine or human interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS) as priming agents.
Main Results:
- Laboratory-prepared zymosans induced equal or significantly greater CL than commercial zymosan in human leukocytes and mouse spleen cells.
- Primed neutrophils showed greatly increased CL with laboratory-prepared zymosans compared to the commercial preparation.
- Reproducible CL responses were observed across different phagocytic cell populations.
Conclusions:
- Effective zymosans for inducing strong and reproducible CL responses can be readily prepared in the laboratory.
- Laboratory-generated zymosans offer a viable alternative to commercial preparations for phagocytic cell activation studies.
- The choice of zymosan preparation can significantly impact the outcome of CL assays.