Al adjuvants can be tracked in viable cells by lumogallion staining

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Al adjuvants can be tracked in viable cells by lumogallion staining

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Publication Article, peer reviewed scientific
Title Al adjuvants can be tracked in viable cells by lumogallion staining
Author(s) Mile, Irene ; Svensson, Andreas ; Darabi, Anna ; Mold, Matthew ; Siesjö, Peter ; Eriksson, Håkan
Date 2015
English abstract
The mechanism behind the adjuvant effect of aluminum salts is poorly understood notwithstanding that aluminum salts have been used for decades in clinical vaccines. In an aqueous environment and at a nearly neutral pH, the aluminumsalts form particulate aggregates, and one plausible explanation of the lack of information regarding the mechanisms could be the absence of an efficientmethod of tracking phagocytosed aluminum adjuvants and thereby the intracellular location of the adjuvant. In this paper, we want to report upon the use of lumogallion staining enabling the detection of phagocytosed aluminum adjuvants inside viable cells. Including micromolar concentrations of lumogallion in the culture medium resulted in a strong fluorescence signal from cells that had phagocytosed the aluminum adjuvant. The fluorescence appeared as spots in the cytoplasm and by confocal microscopy and co-staining with probes presenting fluorescence in the far-red region of the spectrum, aluminum adjuvants could to a certain extent be identified as localized in acidic vesicles, i.e., lysosomes. Staining and detection of intracellular aluminum adjuvants was achieved not only by diffusion of lumogallion into the cytoplasm, thereby highlighting the presence of the adjuvant, but also by pre-staining the aluminum adjuvant prior to incubation with cells. Pre-staining of aluminum adjuvants resulted in bright fluorescent particulate aggregates that remained fluorescent for weeks and with only a minor reduction of fluorescence upon extensive washing or incubation with cells. Both aluminum oxyhydroxide and aluminum hydroxyphosphate, two of the most commonly used aluminum adjuvants in clinical vaccines, could be pre-stained with lumogallion and were easily tracked intracellularly after incubation with phagocytosing cells. Staining of viable cells using lumogallion will be a useful method in investigations of the mechanisms behind aluminum adjuvants' differentiation of antigen-presenting cells into inflammatory cells. Information will be gained regarding the phagosomal pathways and the events inside the phagosomes, and thereby the ultimate fate of phagocytosed aluminum adjuvants could be resolved.
DOI http://dx.doi.org/10.1016/j.jim.2015.04.008 (link to publisher's fulltext)
Publisher Elsevier
Host/Issue Journal of Immunological Methods;
Volume 422
ISSN 0022-1759
Pages 87-94
Language eng (iso)
Subject(s) Aluminum adjuvant
Lumogallion
Flow cytometry
Phagolysosome
Medicine
Research Subject Categories::NATURAL SCIENCES
Handle http://hdl.handle.net/2043/19498 (link to this page)

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