Evaluation of Stable LifeAct-mRuby2- and LAMP1-NeonGreen Expressing A549 Cell Lines for Investigation of Aspergillus fumigatus Interaction with Pulmonary Cells

International Journal of Molecular Sciences

Heidi Oberhauser¹  Natalia Schiefermeier-Mach¹  Lea Heinrich¹  Violetta Moresco¹  Susanne Perkhofer¹  Lukas Lechner¹  Stephan Geley² 

[1] FH Gesundheit/Health University of Applied Sciences Tyrol, Innrain 98, 6020 Innsbruck, Austria;Institute of Pathophysiology, Medical University of Innsbruck, Innrain 52, 6020 Innsbruck, Austria;

关键词: LifeAct;    LAMP1;    stable cell lines;    microscopy;    live-cell imaging;    Aspergillus fumigatus;   

DOI  :  10.3390/ijms22115965

来源: DOAJ

【 摘 要 】

Inhaled Aspergillus fumigatus spores can be internalized by alveolar type II cells. Cell lines stably expressing fluorescently labeled components of endocytic pathway enable investigations of intracellular organization during conidia internalization and measurement of the process kinetics. The goal of this report was to evaluate the methodological appliance of cell lines for studying fungal conidia internalization. We have generated A549 cell lines stably expressing fluorescently labeled actin (LifeAct-mRuby2) and late endosomal protein (LAMP1-NeonGreen) following an evaluation of cell-pathogen interactions in live and fixed cells. Our data show that the LAMP1-NeonGreen cell line can be used to visualize conidia co-localization with LAMP1 in live and fixed cells. However, caution is necessary when using LifeAct-mRuby2-cell lines as it may affect the conidia internalization dynamics.

【 授权许可】

Unknown