A rapid evaluation of phagocytosis and killing of Candida albicans by CD13 + leukocytes

M. Saresella, K. Roda, L. Speciale, D. Taramelli, E. Mendozzi, F. Guerini, P. Ferrante

Research output: Contribution to journalArticle

Abstract

Flow cytometry can be adopted for routine monitoring of the immune functions of human polymorphonuclear leukocytes (PMNs) in several disease states. We recently developed a rapid and reproducible assay for the evaluation of the phagocytosis and killing of Candida albicans blastospores by human PMNs. Whole blood leukocytes were incubated with opsonized fluorescein isothiocyanate-labeled (FITC-labeled) blastospores for phagocytosis and killing assays. To discriminate between ingested, membrane- bound and free C. albicans blastospores, ethidium bromide (EtBr) was added to the samples prior to the flow cytometric analysis. EtBr induces a loss of green fluorescence in non-phagocytized C. albicans blastospores. Phagocytosis is determined by gating the phagocytes and calculating the percentage of phagocyte-associated green fluorescent cells. Intracellular killing is determined by first lysing phagocytes by hypotonic shock and then adding propidium iodide (PI) in order to identify red dead blastospores. Killing is measured in terms of the percentage of double-marked blastospore cells. We suggest that this method is a reliable and inexpensive technique to evaluate the immune reactivity of PMNs and peripheral blood monocytes (PBMs) in cases of immunosuppression.

Original languageEnglish
Pages (from-to)227-234
Number of pages8
JournalJournal of Immunological Methods
Volume210
Issue number2
DOIs
Publication statusPublished - Dec 29 1997

Keywords

  • Adhesion
  • Cytotoxicity
  • Flow cytometry
  • Fluorescent Candida albicans

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Fingerprint Dive into the research topics of 'A rapid evaluation of phagocytosis and killing of Candida albicans by CD13 + leukocytes'. Together they form a unique fingerprint.

  • Cite this