Premium
Fast and Quantitative Evaluation of Human Leukocyte Interaction with Aspergillus fumigatus Conidia by Flow Cytometry
Author(s) -
Hartung Susann,
Rauh Christopher,
Hoang Thi Ngoc Mai,
Jahreis Susanne,
Wagner Kathleen,
Macheleidt Juliane,
Brakhage Axel A.,
Rummler Silke,
Hochhaus Andreas,
LilienfeldToal Marie
Publication year - 2019
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.23653
Subject(s) - aspergillus fumigatus , flow cytometry , conidium , microbiology and biotechnology , cytometry , biology , chemistry , immunology , botany
Systemic infections with the opportunistic mold Aspergillus fumigatus are a great threat to immunocompromised patients such as transplant recipients. Immunological research on A. 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 rates of A. fumigatus conidia in different stages and after several incubation times using this method. Moreover, we provide an example of application by comparing phagocytosis of A. fumigatus mutants to the wild type. © 2018 International Society for Advancement of Cytometry