Fast and Quantitative Evaluation of Human Leukocyte Interaction with Aspergillus fumigatus Conidia by Flow Cytometry

AbstractSystemic infections with the opportunistic moldAspergillus fumigatus are a great threat to immunocompromised patients such as transplant recipients. Immunological research onA. fumigatus involves the measurement of phagocytosis of fungal conidia (spores) by human phagocytes. Here, we present a fast and flexible way to analyze phagocytosis by flow cytometry using fluorescein isothiocyanate (FITC) labeling of conidia prior to co ‐incubation with human leukocytes and an anti‐FITC counterstaining step postincubation to allow the discrimination of internalized and adherent conidia. In contrast to many other protocols, this method can be combined with further surface marker analyses. We sought to determine phagocytosis rate s ofA. fumigatus conidia in different stages and after several incubation times using this method. Moreover, we provide an example of application by comparing phagocytosis ofA. fumigatus mutants to the wild type. © 2018 International Society for Advancement of Cytometry
Source: Cytometry Part A - Category: Molecular Biology Authors: Tags: Technical Note Source Type: research