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Involvement of phospholipase A2 activation in anthrax lethal toxin-induced cytotoxicity

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Abstract

The molecular mechanism of cytotoxic effect exerted by the lethal toxin (LeTx) of Bacillus anthracis is not well understood. In the present study, using primary culture of mouse peritoneal macrophages, we have investigated possible cytotoxic mechanisms. LeTx was not found to induce high levels of nitric oxide (NO) production for NO-mediated toxicity. Fragmentation of DNA, a biochemical marker of apoptosis, was not observed in LeTx-treated cells. Pretreatment of cells with antioxidants such as melatonin and dehydroepiandrosterone (DHEA) did not protect the LeTx-induced cytotoxicity. However, addition of phospholipase A2 (PLA2) inhibitors (quinacrine, p-bromophenacyl bromide, manoalide, butacaine) to the culture medium resulted in the inhibition of cytotoxicity of LeTx in a dose-dependent manner. LeTx-induced cytotoxicity was also inhibited by the tyrosine-specific protein kinase inhibitor genistein, but not by the protein kinase C inhibitors staurosporine or H-7. The results of these studies indicate a role for PLA2 and protein kinase in the cytotoxic mechanism of macrophages by anthrax lethal toxin.

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References

  • Akiyama T, Ishida J, Nagagawa S. Genistein, a specific inhibitor of tyrosine-specific protein kinases. J Biol Chem. 1987;262:5592-5.

    Google Scholar 

  • Albina JE, Cui S, Mateo RB, Reichner S. Nitric oxide-mediated apoptosis in murine peritoneal macrophages. J Immunol. 1993;150:5080-5.

    Google Scholar 

  • Araghiniknam M, Chung S, Nelson-White T, Eskelson C, Watson RR. Antioxidant activity of dioscorea and dehydroepiandrosterone in older humans. Life Sci. 1996;59:147-57.

    Google Scholar 

  • Balsinde J, Dennis EA. Distinct roles in signal transduction for each of the phospholipase A2 enzymes present in P388D1 macrophages. J Biol Chem. 1996;271:6758-65.

    Google Scholar 

  • Balsinde J, Barbour SE, Bianco ID, Dennis EA. Arachidonic acid mobilization in P388D1 macrophages is controlled by two distinct Ca2+-dependent phospholipase A2 enzymes. Proc Natl Acad Sci USA. 1994;91:11060-4.

    Google Scholar 

  • Bhatnagar R, Friedlander AM. Protein synthesis is required for expression of anthrax lethal toxin cytotoxicity. Infect Immun. 1994;62:2958-62.

    Google Scholar 

  • Bhatnagar R, Singh Y, Leppla SH, Friedlander AM. Calcium is required for the expression of anthrax lethal toxin activity in the macrophage-like cell line J774A.1. Infect Immun. 1989; 57:2107-14.

    Google Scholar 

  • Bonfoco E, Kraine D, Ankarcrona M, Nicotera P, Lipton S. Apoptosis and necrosis: two distinct events induced, respectively, by mild and intense insults with N-methyl-D-aspartate or nitric oxide/superoxide in cortical cell cultures. Proc Natl Acad Sci USA. 1995;92:7162-6.

    Google Scholar 

  • Choi DS, Shin S, Kim YB, Cha SH, Sok DE. An additional mechanism for the cytotoxicity of 2-chlororthylethyl sulfide in spleen lymphocytes. J Biochem Mol Biol. 1995;28:79-82.

    Google Scholar 

  • Danenberg HD, Ben-Yehuda A, Zakay-Rones Z, Friedman G. Dehydroepiandrosterone treatment reverses the impaired immune response of old mice to influenza vaccination and protects from influenza infection. Vaccine. 1995;13:1445-8.

    Google Scholar 

  • Dennis EA. Diversity of group types, regulation, and function of phospholipase A2. J Biol Chem. 1994;269:13057-60.

    Google Scholar 

  • Duan H, Chinnaiyan AM, Hudson PL, Wing JP, He W, Dixit VM. ICE-LAP3, a novel mammalian homologue of the Caenorhabditis elegans cell death protein ced-3 is activated during Fas-and tumor necrosis factor-induced apoptosis. J Biol Chem. 1996;271:1621-5.

    Google Scholar 

  • Fink DW, Guroff G. Nerve growth factor stimulation of arachidonic acid release from PC12 cells: independence from phosphoinositide turnover. J Neurochem. 1990;55:1716-26.

    Google Scholar 

  • Friedlander AM. Macrophages are sensitive to anthrax lethal toxin through an acid-dependent process. J Biol Chem. 1986;261:7123-6.

    Google Scholar 

  • Glaser KB, Asmis R, Dennis EA. Bacterial lipopolysaccharide priming of P388D1 macrophage-like cells for enhanced arachidonic acid metabolism. J Biol Chem. 1990;265:8658-64.

    Google Scholar 

  • Goldberg HJ, Viegas MM, Margolis BL, Schlessinger J, Skorecki KL. The tyrosine kinase activity of the epidermal-growth-factor receptor is necessary for phospholipase A2 activation. Biochem J. 1990;267:461-5.

    Google Scholar 

  • Green LC, Wagner DR, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and [15N]-nitrate in biological fluids. Anal Biochem. 1982;126:131-8.

    Google Scholar 

  • Hale AJ, Smith CA, Sutherland LC. Apoptosis: molecular regulation of cell death. Eur J Biochem. 1996;236:1-26.

    Google Scholar 

  • Hanna PC, Acosta D, Collier RJ. On the role of macrophages in anthrax. Proc Natl Acad Sci USA. 1993;90:10198-201.

    Google Scholar 

  • Hanna PC, Kruskal BA, Ezekowitz AB, Bloom BR, Collier RJ. Role of macrophage oxidative burst in the action of anthrax lethal toxin. Mol Med. 1994;1:7-18.

    Google Scholar 

  • Hazen SL, Zupan LA, Weiss RH, Getman DP, Gross RW. Suicide inhibition of canine myocardial cytosolic calcium-independent phospholipase A2. J Biol Chem. 1991;266:7227-32.

    Google Scholar 

  • Hofmann SL, Prescott SM, Majerus PW. The effects of mepacrine and p-bromophenacyl bromide on arachidonic acid release in human platelets. Arch Biochem Biophys. 1982; 215:237-44.

    Google Scholar 

  • Kawamoto S, Hidaka H. 1-(-5-Isoquinolinesulfonyl)-2-methyl-piperazine (H-7) is a selective inhibitor of protein kinase C in rabbit platelet. Biochem Biophys Res Commun. 1984; 125:258-64.

    Google Scholar 

  • Klimpel KR, Arora M, Leppla SH. Anthrax toxin lethal factor contains a zinc metalloprotease consensus sequence which is required for lethal toxin activity. Mol Microbiol. 1994;13:1093-100.

    Google Scholar 

  • Kolb H, Kolb-Bachofen V. Nitric oxide: a pathogenetic factor in autoimmunity. Immunol Today. 1992;13:157-62.

    Google Scholar 

  • Leppla SH. Bacillus anthracis calmodulin dependent adenyl cyclase: chemical and enzymatic properties and interaction with eucaryotic cells. Cyclic Nucleo Protein Phosphoryl Res. 1984;19:189-98.

    Google Scholar 

  • Leppla SH. Production and purification of anthrax toxin. Methods Enzymol. 1988;165:103-16.

    Google Scholar 

  • Leppla SH. The anthrax toxin complex. In: Alouf J, ed. Sourcebook of bacterial protein toxins. New York: Academic Press; 1991:277-302.

    Google Scholar 

  • Menard A, Papini E, Mock M, Montecucco C. The cytotoxic activity of Bacillus anthracis lethal factor in inhibited by leukotriene A4 hydrolase and metallopeptidase inhibitors. Biochem J. 1996;320:687-91.

    Google Scholar 

  • Meyer T, Regenass U, Fabbro D. A derivative of staurosporine (CGP 41 251) shows selectivity for protein kinase C inhibition and in vitro anti-proliferative as well as in vivo antitumor activity. Int J Cancer. 1989;43:851-6.

    Google Scholar 

  • Moncada S, Palmer RMJ, Higgs EA. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev. 1991; 43:109-42.

    Google Scholar 

  • Mukherjee AB, Miele L, Pattabiraman N. Phospholipase A2 enzymes: regulation and physiological role. Biochem Pharmacol. 1994;48:1-10.

    Google Scholar 

  • Nathan CF. Nitric oxide as a secretory product of mammalian cells. FASEB J. 1992;6:3051-64.

    Google Scholar 

  • Poeggeler B, Reiter RJ, Tan DX, Chen LD, Manchester LC. Melatonin, hydroxyl radical-mediated oxidative damage and aging: a hypothesis. J Pineal Res. 1993;14:151-68.

    Google Scholar 

  • Pozo D, Reiter RJ, Calvo JP, Guerrero JM. Physiological concentrations of melatonin inhibit nitric oxide synthase in rat cerebellum. Life Sci. 1994;55:PL455-60.

    Google Scholar 

  • Ray P, Berman JD, Middleton W, Brendle J. Botulinum toxin inhibits arachidonic acid release associated with acetylcholine release from PC12 cells. J Biol Chem. 1993;268:11057-64.

    Google Scholar 

  • Reiter RJ, Melchiorri D, Sewerynek E. A review of the evidence supporting melatonin's role as an antioxidant. J Pineal Res. 1995;18:1-11.

    Google Scholar 

  • Segal AW, Abo A. The biochemical basis of the NADPH oxidase of phagocytes. Trends Biochem Sci. 1993;18:43-7.

    Google Scholar 

  • Stephen J. Anthrax toxin. In: Doner F, Drews J, eds. Pharmacology of bacterial protein toxins. Oxford: Pergamon Press; 1986:381-95.

    Google Scholar 

  • Street IP, Lin HK, Laliberte F. Slow and tight-binding inhibitors of the 85-kDa human phospholipase A2. Biochemistry. 1993;32:5935-40.

    Google Scholar 

  • Ueda N, Shah S. Apoptosis. J Lab Clin Med. 1994;124:169-77.

    Google Scholar 

  • Wheeler LA, Sachs G, DeVries G, Goodrum D, Woldemussie E, Muallen S. Manoalide, a natural sesterterpenoid that inhibits calcium channels. J Biol Chem. 1987;262:6531-8.

    Google Scholar 

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  1. CBR Department, Agency for Defense Development, Yuseong, Taejon, Korea

    S. Shin, Y.-B. Kim & G.-H. Hur

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  1. S. Shin
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  2. Y.-B. Kim
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  3. G.-H. Hur
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Shin, S., Kim, YB. & Hur, GH. Involvement of phospholipase A2 activation in anthrax lethal toxin-induced cytotoxicity. Cell Biol Toxicol 15, 19–29 (1999). https://doi.org/10.1023/A:1007546505528

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