Journal
EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES
Volume 27, Issue 2-3, Pages 252-258Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.ejps.2005.10.006
Keywords
HL-60; chemiluminescence; ROS; yeast; lipopolysaccharide; lipoteichoic acid; pyrogen assay
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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. Further-more, the utilization of the assay for pyrogenic examination of recombinant drugs derived from yeast expression systems would be relevant to examine. (C) 2005 Elsevier B.V. All rights reserved.
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