The use of human monocytoid lines as indicators of endotoxin

Details

Serval ID
serval:BIB_9CBF27FE3CCD
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
The use of human monocytoid lines as indicators of endotoxin
Journal
Journal of Immunological Methods
Author(s)
Eperon  S., Jungi  T. W.
ISSN
0022-1759
Publication state
Published
Issued date
08/1996
Peer-reviewed
Oui
Volume
194
Number
2
Pages
121-9
Notes
Comparative Study
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Aug 14
Abstract
We wish to develop an in vitro test system for pyrogenic substances. A major source of pyrogen activity is endotoxin. Here we describe a highly sensitive endotoxin-monitoring system based on cytokine measurement in human cell lines of myelomonocytoid origin. The following measures were taken to develop an endotoxin monitoring system of high sensitivity. (i) Mono Mac 6 (MM6) and THP-1 cells, which both represent advanced stages of myelomonocytic development, were better suited as endotoxin indicators than the more immature U-937 line. (ii) In order to enhance cell surface expression of CD14, a major lipopolysaccharide (LPS) receptor, cells were pretreated for 2 days with calcitriol. (iii) The use of fetal calf serum (FCS) without detectable endotoxin traces was essential for maintaining a high LPS sensitivity. (iv) Selected subclones of either THP-1 or MM6 were significantly more sensitive LPS indicators than bulk cultures from which the clones originated. (v) Based on stimulation indices, a commercial tumor necrosis factor-alpha (TNF-alpha) immunoassay proved to be a more sensitive LPS indicator than other cytokine assays or the expression of procoagulant activity/tissue factor. Thus we were able to eliminate the disadvantage of previous cell line-based systems (i.e., low sensitivity) without loss of reproducibility which is seen when using fresh blood, monocytes or monocyte-derived macrophages. High endotoxin sensitivity is a prerequisite for a test system specifically indicating pyrogen activity, because it permits the testing of substances at higher dilutions, thereby minimizing nonspecific interference.
Keywords
Calcitriol/pharmacology Cell Differentiation/drug effects/physiology Cell Division/drug effects/physiology Cell Line Coagulants/analysis/pharmacology Drug Evaluation, Preclinical/methods Endotoxins/*analysis Humans Limulus Test Lipopolysaccharides/pharmacology Macrophages/cytology/drug effects Monocytes/*cytology/*drug effects Phenotype Pyrogens/analysis Reproducibility of Results Sensitivity and Specificity Tumor Necrosis Factor-alpha/analysis
Pubmed
Web of science
Create date
17/01/2008 13:27
Last modification date
20/08/2019 15:03
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