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

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 fas...

Full description

Saved in:
Bibliographic Details
Published in:Cytometry. Part A 2019-03, Vol.95 (3), p.332-338
Main Authors: Hartung, Susann, Rauh, Christopher, Hoang, Thi Ngoc Mai, Jahreis, Susanne, Wagner, Kathleen, Macheleidt, Juliane, Brakhage, Axel A., Rummler, Silke, Hochhaus, Andreas, Lilienfeld‐Toal, Marie
Format: Article
Language:English
Subjects:
anti‐FITC antibody
Aspergillosis - immunology
Aspergillus fumigatus
Aspergillus fumigatus - genetics
Aspergillus fumigatus - growth & development
Aspergillus fumigatus - metabolism
Conidia
FITC
Flow cytometry
Flow Cytometry - methods
Fluorescein
Fluorescein isothiocyanate
Fluorescein-5-isothiocyanate - analysis
Fluorescein-5-isothiocyanate - chemistry
Fluorescence
Humans
Immunocompromised hosts
Immunology
Leukocytes
Leukocytes - immunology
Leukocytes - microbiology
Municipalities
Mutants
Phagocytes
Phagocytosis
Phagocytosis - genetics
Phagocytosis - immunology
Quantitative analysis
Spores
Spores, Fungal - metabolism
Surface markers
Yard waste
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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
ISSN:1552-4922
1552-4930
DOI:10.1002/cyto.a.23653