Lipopolysaccharide (LPS) and zymosan-resistant mutant isolated from a macrophage-like cell line, WEHI-3, with a defective response to LPS under serum-free conditions.
A LPS-resistant mutant, W3SF-1, was isolated from a murine macrophage-like cell line, WEHI-3. The W3SF-1 mutant did not produce a significant amount of nitric oxide (NO) or TNF-alpha even with high concentrations of LPS in the presence or absence of FCS, whereas the parental WEHI-3 cells produced them in response to LPS. The parental cells expressed a significant level of TNF-alpha mRNA after LPS stimulation, whereas the mutant cells did not. This defective response of the mutant cells to LPS was neither dependent on the concentration or chemical structure of LPS, nor on the time of LPS treatment. The mutant cells also showed a defective response to zymosan, suggesting that the defect in the mutant cells is common to LPS and zymosan in the signal transduction pathways. The parental and mutant cells showed similar levels of Mac1, F4/80 and CD14, suggesting that these surface markers of macrophages are not linked directly to the defective responses of the mutant to LPS. The treatment of mutant cells with IFN-gamma did not restore the defect of NO or TNF-alpha production on LPS treatment. Binding experiments with 125I-labelled LPS showed a similar binding affinity for LPS in the parental and the mutant cells. These results suggest that the defect in the W3SF-1 mutant cells may not reside in the LPS binding but rather in the early step of signal transduction pathways in the cells after LPS binding.
Ohki
K
K
Department of Microbiology, Saga Medical School, Saga, Japan. ohki@smsnet.saga-med.ac.jp
Amano
F
F
Kohashi
O
O
D000818:Animals; D002460:Cell Line; D016895:Culture Media, Serum-Free; D005434:Flow Cytometry; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D008050:Lipid A / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008262:Macrophage Activation; D008264:Macrophages / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism*; D015054:Zymosan / Q000494:pharmacology*
D016895:Culture Media, Serum-Free; D008050:Lipid A; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D007371:Interferon-gamma; D015054:Zymosan