title: A potential test system for detecting contaminations by bacterial lipoproteins.
abstract: Biological specimens are often contaminated with bacteria-derived products such as LPS or lipoproteins (LP), which trigger unwanted inflammatory responses in hosts. Whereas a contamination by LPS can be determined by various test systems, a contamination by LP can as yet not be determined. TLR4 and TLR2 are key components of the LPS and the LP receptor complex, respectively. It was tested in this study whether HEK293 cell stably transfected with bovine TLR2 have the ability to react to low concentrations of diacylated and triacylated synthetic LP. The stable cell lines we present here recognize low concentrations of synthetic LP resembling LP of different bacteria. Therefore, these cells are suitable to detect low contaminations present in probes. For example, HEK293 cells stably transfected with bovine TLR2 recognized an egg albumin preparation as contaminated, as evidenced by copious production of IL-8. In contrast, these cells did not respond to a highly purified human serum albumin (HSA) preparation used in the clinic but responded to HSA containing small amounts of diacylated synthetic LP. The TLR4 ligand LPS is often said to activate TLR2. Here we present evidence that LP contaminations are responsible for TLR2 activity. HEK293 cells stably transfected with bovine TLR2 and TLR1 (e.g. clone 1) did not respond to ultra-pure Escherichia coli LPS preparations but acquired responsiveness when stimulated with differently purified commercial LPS. Thus, the described system involving HEK293 cells stably transfected with bovine TLR2 and TLR1 is the first test system described attempting to measure a contamination by LP.
authors: Department of Immunology and Cell Biology, Research Center Borstel, 23845 Borstel, Germany.
authors: Katja
authors: K
authors: Farhat
authors: Artur J
authors: AJ
authors: Ulmer
authors: Thomas W
authors: TW
authors: Jungi
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis*; D002417:Cattle; D002999:Clone Cells / Q000737:chemistry; D057809:HEK293 Cells / Q000737:chemistry / Q000382:microbiology; D006801:Humans; D016209:Interleukin-8 / Q000032:analysis; D008070:Lipopolysaccharides / Q000032:analysis; D008074:Lipoproteins / Q000032:analysis*; D012709:Serum Albumin / Q000032:analysis; D051194:Toll-Like Receptor 1 / Q000032:analysis; D051195:Toll-Like Receptor 2 / Q000032:analysis; D051197:Toll-Like Receptor 4 / Q000032:analysis; D014162:Transfection / Q000379:methods
chemical_list: D001426:Bacterial Proteins; D016209:Interleukin-8; D008070:Lipopolysaccharides; D008074:Lipoproteins; D012709:Serum Albumin; D051194:Toll-Like Receptor 1; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4