The use of human monocytoid lines as indicators of endotoxin
Détails
ID Serval
serval:BIB_9CBF27FE3CCD
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
The use of human monocytoid lines as indicators of endotoxin
Périodique
Journal of Immunological Methods
ISSN
0022-1759
Statut éditorial
Publié
Date de publication
08/1996
Peer-reviewed
Oui
Volume
194
Numéro
2
Pages
121-9
Notes
Comparative Study
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Aug 14
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Aug 14
Résumé
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.
Mots-clé
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
Création de la notice
17/01/2008 14:27
Dernière modification de la notice
20/08/2019 16:03