Human monocytoid cell lines as indicators of endotoxin: comparison with rabbit pyrogen and Limulus amoebocyte lysate assay

Details

Serval ID
serval:BIB_83EE56D6048C
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Human monocytoid cell lines as indicators of endotoxin: comparison with rabbit pyrogen and Limulus amoebocyte lysate assay
Journal
Journal of Immunological Methods
Author(s)
Eperon  S., De Groote  D., Werner-Felmayer  G., Jungi  T. W.
ISSN
0022-1759
Publication state
Published
Issued date
09/1997
Peer-reviewed
Oui
Volume
207
Number
2
Pages
135-45
Notes
Comparative Study
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Sep 24
Abstract
The aim of this study was to develop an in vitro test system for pyrogenic substances. Three clones derived from human monocytoid cell lines, which were selected by their high sensitivity to lipopolysaccharide (LPS), were assessed for tumor necrosis factor (TNF) production. Their response to pyrogen-containing samples was compared with that in a Limulus amoebocyte lysate assay and the rabbit pyrogen test. We show here that the induction of TNF in these clones is a valid in vitro alternative to determine endotoxin in commercial preparations requiring pyrogenicity testing. Cell clones derived from Mono Mac 6 (MM6 2H8 and MM6 4B5) responded to sub-ng/ml concentrations of complete rough-strain and smooth-strain LPS, to ng/ml concentrations of diphosphoryl-lipid A, and to microgram/ml concentrations of monophosphoryl-lipid A and to detoxified LPS. Cells reacted to > or = 1 microgram/ml lipoteichoic acid by TNF production, and were relatively insensitive to toxic shock syndrome toxin-1 (TSST-1) and to muramyl dipeptide adjuvant peptide. The reaction pattern of a clone derived from THP-1 (THP-1 1G3) was in general, similar to that of the MM6 clones, except that THP-1 1G3 failed to react to diphosphoryl-lipid A. When tested on commercial samples destined for parenteral use, there was a close correlation between a sensitive Limulus amoebocyte lysate (LAL) test and the cell culture test on the one hand, and between the pyrogen test and the cell culture test on the other hand. The data suggest that this cell-based test is able to recognize pyrogens derived from gram-negative organisms in test samples with appropriate sensitivity and specificity. This test appears to be able to eliminate some of the false-positive data obtained in the LAL test.
Keywords
Acetylmuramyl-Alanyl-Isoglutamine/toxicity Animals Bacterial Toxins/toxicity Bacterial Vaccines/adverse effects Biological Assay/*methods Clone Cells Endotoxins/*analysis Enterotoxins/toxicity False Positive Reactions Humans Limulus Test Lipid A/*toxicity Lipopolysaccharides/*toxicity Monocytes/*drug effects/metabolism Pyrogens/*analysis Rabbits Sensitivity and Specificity *Superantigens Teichoic Acids/toxicity Tumor Necrosis Factor-alpha/*metabolism
Pubmed
Web of science
Create date
17/01/2008 13:27
Last modification date
20/08/2019 14:43
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