Rapid damage to the cells was observed when a macrophage-like cell line, J774.1, was incubated with LPS in the presence of cycloheximide (CHI). Cells of an LPS-sensitive subline of J774.1, JA-4, but not an LPS-resistant mutant, LPS1916, became swollen and detached from the culture dish on treatment with 10 ng/ml LPS and 10 μg/ml CHI at 37°C for 4 h, accompanied with the release of lactate dehydrogenase. These changes were dependent on both LPS and CHI, and the addition of CHI to the macrophages later than 1 h after the addition of LPS failed to induce the cytotoxicity. As for the mechanism, protein kinase C (PKC) seemed to be involved, because down-regulation of PKC and staurosporine reduced the cytotoxicity. The cytotoxicity was dependent on serum, suggesting the involvement of CD14/LPS binding protein. However, TNFα does not seem to participate, because an anti-TNFα antibody did not inhibit the cytotoxicity and TNFα could not replace LPS.
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References
1.
Morrison D., Ryan J.L.Bacterial endotoxins and host immune responses. Adv Immunol1979; 28: 293-450.
2.
Virca G.D., Kim S.Y., Glaser K.B., Ulevitch R.J.Lipopolysaccharide induces hyporesponsiveness to its own action in RAW264.7 cells. J Biol Chem1989; 264: 21951-21956.
3.
Heumann D., Gallay P., Barras C. et al. Control of lipopolysaccharide (LPS) binding and LPS-induced tumor necrosis factor secretion in human peripheral blood monocytes. J Immunol1992; 148: 3505-3512.
4.
Raetz C.R.H. , Ulevitch R.J., Wright S.D., Sibley C.H., Ding A., Nathan C.F.Gram-negative endotoxin: an extraordinary lipid with profound effects on eukaryotic signal transduction. FASEB J1991; 5: 2652-2660.
5.
Johnson W.J., Adams D.O.Activation of mononuclear phagocytes for tumor cytolysis: analysis of inductive and regulatory signals. In: Volkman A. ed. Mononuclear Phagocyte Biology. New York: Marcel Dekker, 1984; 279-300.
6.
Amano F., Akamatsu Y.A lipopolysaccharide (LPS)-resistant mutant isolated from a macrophage-like cell line, J774.1, exhibits an altered activated macrophage phenotype in response to LPS. Infect Immun1991; 59: 2166-2174.
7.
Hara-Kuge S. , Amano F., Nishijima M., Akamatsu Y.Isolation of an LPS-resistant mutant, with defective LPS binding, of cultured macrophage-like cells. J Biol Chem1990; 265: 6606-6610.
8.
Amano F., Fujita T., Yamamoto S., Kurata T. Manuscript in preparation.
9.
Nishijima M. , Hara-Kuge S., Takasuka N. et al. Identification of a biochemical lesion, and characteristic response to lipopolysaccharide (LPS) of a cultured macrophage-like cell mutant with defective LPS-binding. J Biochem1994; 116: 1082-1087.
10.
Amano F., Akamatsu Y.Reversal by IFN-γ of the defect in LPS-induced O2--generating activity in an LPS-resistant mutant. J Pharmacodyn1992; 15: S.62.
11.
Inoue S., Itagaki S-I., Amano F.Intracellular killing of Listeria monocytogenes in the J774.1 macrophage-like cell line and the lipopolysaccharide (LPS)-resistant mutant LPS1916 cell line defective in the generation of reactive oxygen intermediates after LPS treatment . Infect Immun1995; 63: 1876-1886.
12.
Matic M., Simon S.R.Tumor necrosis factor release from lipopolysaccharide-stimulated human monocyte: lipopolysaccharide tolerance in vitro. Cytokine1991; 2: 576-583.
13.
Sarih M., Souvannavong V., Adam A.Nitric oxide synthase induces macrophage death by apoptosis. Biochem Biophys Res Commun1993; 191: 503-508.
14.
Amano F., Noda T.Improved detection of nitric oxide radical (NO°) production in an activated macrophage culture with a radical scavenger, carboxy PTIO, and Griess reagent . FEBS Lett1995; 368: 425-428.
15.
Pesce A., McKay R.H., Stolzenbach F. et al. The comparative enzymology of lactic dehydrogenases. J Biol Chem1964; 239: 1753-1761.
16.
Fujihara M. , Connonly N., Ito N. et al. Properties of protein kinase C isoforms (βII, ε, and ζ) in a macrophage cell line (J774) and their roles in LPS-induced nitric oxide production. J Immunol1994; 152: 1898-1906.
17.
Beutler B., Milsark I.W., Cerami A.C.Passive immunization against cachectin/tumor necrosis factor protects mice from lethal effect of endotoxin. Science1985 ; 229: 869-871.
18.
Tracey K., Beutler B., Lowry S.F. et al. Shock and tissue injury induced by recombinant human cachectin . Science1986; 234: 470-474.
19.
Haziot A., Ferrero E., Koentgen F. et al. Resistance to endotoxin shock and reduced dissemination of Gram-negative bacteria in CD14-deficient mice. Immunity1996; 4: 407-414.
20.
Delude R.L., Savedra Jr R., Zhao H. et al. CD 14 enhances cellular responses to endotoxin without impairing ligand-specific recognition. Proc Natl Acad Sci USA1995; 92: 9288-9292.
21.
Kielian T.L. , Blecha F.CD 14 and other recognition molecules for lipopolysaccharide: a review. Immunopharmacology1995; 29: 187-205.
22.
Kirikae F., Kirikae T., Qureshi N. et al. CD14 is not involved in Rhodobacter sphaeroides diphosphoryl lipid A inhibition of tumor necrosis factor alfa and nitric oxide induction by taxol in murine macrophages. Infect Immun1995; 63: 486-497.
