A new technique, requiring small amounts of cells, for the parallel study of chemiluminescence and phagocytosis via different receptors in the same cell population.
An assay permitting the parallel assessment of phagocytosis and chemiluminescence in the same cell population has been developed. The method is based on the phagocytosis of fluorescein isothiocyanate-conjugated yeast particles, either unopsonized or opsonized with complement factor C3 or IgG, by purified cells in suspension in a luminometer. Only a small number of cells (2 x 10(4)-1 x 10(5)) is required, and the reproducibility is high. Moreover, the technique permits phagocytosis to be related to oxygen-dependent killing activity in the same cell population. Since phagocytosis, degranulation and oxygen radical formation as a consequence of well-defined receptor recognition mechanisms can be characterized in very small cell populations, the method is suitable for monitoring the phagocytic function of cells from extravascular sites.
Sandgren
C H
CH
Department of Veterinary Medical Chemistry, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Nordling
K
K
Björk
I
I
D000818:Animals; D002417:Cattle; D015550:Cell Degranulation; D003176:Complement C3 / Q000276:immunology; D005260:Female; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005609:Free Radicals; D007074:Immunoglobulin G / Q000276:immunology; D008163:Luminescent Measurements; D009504:Neutrophils / Q000502:physiology*; D010100:Oxygen; D010587:Phagocytosis; D011951:Receptors, Complement / Q000502:physiology; D011961:Receptors, Fc / Q000502:physiology; D015203:Reproducibility of Results; D013861:Thiocyanates; D013997:Time Factors
D003176:Complement C3; D005452:Fluoresceins; D005609:Free Radicals; D007074:Immunoglobulin G; D011951:Receptors, Complement; D011961:Receptors, Fc; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate; D010100:Oxygen