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A Flow Cytometric Method for the Analysis of Phagocytosis and Killing by Polymorphonuclear Leukocytes
Author(s) -
SARESELLA M.,
RODA K.,
SPECIALE L.,
TARAMELLI D.,
MENDOZZI E.,
GUERINI F.,
FERRANTE P.
Publication year - 1997
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.1997.tb46236.x
Subject(s) - phagocytosis , flow cytometry , medicine , library science , immunology , computer science
Phagocytes represent the first line of defense against invasive microorganisms as they are capable of rapid phagocytosis and killing of bacteria and fungi. A rapid evaluation of both the number and function of the human polymorphonuclear leukocyte (PMN) population could be of fundamental importance in defining the immunological defects of patients with recurrent or persistent infections. Moreover, any technical modifications that could improve the clinical screening of PMNs function would be advantageous for the treatment of many disease states. Flow cytometry can be adopted for routine monitoring of the immune functions of human polymorphonuclear leukocytes (PMNs) in several disease states.I4 In this study, we describe a fast, reliable, and inexpensive method for studying the phagocytosis and killing of Candida albicans blastospores by fresh human polymorphonuclear cells, using flow cytometry (FCM). We have modified previously described methodss in order to achieve the reproducibility and speed necessary for testing several samples in the diagnostic laboratory. This assay is capable of distinguishing adherent blastospores from those that have been ingested and permits an evaluation of intracellular killing as well. Briefly summarized, the advantages offered by this assay are ( I ) more rapid PMN preparation; (2) the staining procedure involves less time, thereby contributing to a faster assay; (3) the C. albicans to PMN ratio is 2: 1, which allows an accurate estimation of truly phagocytic PMN; (4) fluorescencelabeled C. albicans blastospores can be frozen and used when needed without loss of fluorescence; and (5 ) this assay also measures killing. The technique can be easily