Utilization of the human cell line HL-60 for chemiluminescence based detection of microorganisms and related substances.
In this paper we describe a new pyrogen assay using the human leukemia cell line HL-60. The cell line is differentiated using all-trans retinoic acid (ATRA) to generate a cell population that resembles mature granulocytes. The differentiated HL-60 cell is capable of generating reactive oxygen species (ROS) when challenged with pyrogenic substances. In a luminol enhanced chemilumimetric assay the responsiveness of differentiated HL-60 cells is tested towards Salmonella typhimurium, Bacillus subtilis, Saccharomyces cerevisiae, Candida albicans, lipopolysaccharide (LPS) and lipoteichoic acid (LTA). The results show a poor sensitivity to S. typhimurium but displays good sensitivity towards B. subtilis, LTA and LPS. Furthermore, the sensitivity towards the yeasts C. albicans and S. cerevisiae is considerably better than obtained in other in vitro cell systems. Overall these results indicate that the HL-60 cell assay possibly could be evolved to a supplementary assay for the known pyrogenic detection assays. Furthermore, the utilization of the assay for pyrogenic examination of recombinant drugs derived from yeast expression systems would be relevant to examine.
Timm
Michael
M
Department of Pharmacology, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, 2100 Copenhagen, Denmark.
Hansen
Erik W
EW
Moesby
Lise
L
Christensen
Jens D
JD
D001412:Bacillus subtilis / Q000302:isolation & purification / Q000378:metabolism; D001681:Biological Assay / Q000379:methods*; D002176:Candida albicans / Q000302:isolation & purification / Q000378:metabolism; D002454:Cell Differentiation; D004340:Drug Contamination; D018922:HL-60 Cells / Q000187:drug effects / Q000378:metabolism* / Q000382:microbiology; D006801:Humans; D007202:Indicators and Reagents; D008070:Lipopolysaccharides / Q000494:pharmacology; D049449:Luminescence; D008165:Luminol; D011749:Pyrogens / Q000032:analysis* / Q000302:isolation & purification; D017382:Reactive Oxygen Species / Q000032:analysis / Q000378:metabolism; D015203:Reproducibility of Results; D013682:Teichoic Acids / Q000494:pharmacology; D013997:Time Factors; D014212:Tretinoin
D007202:Indicators and Reagents; D008070:Lipopolysaccharides; D011749:Pyrogens; D017382:Reactive Oxygen Species; D013682:Teichoic Acids; C009900:lipoteichoic acid; D014212:Tretinoin; D008165:Luminol