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. 1997 Jul;65(7):2932–2940. doi: 10.1128/iai.65.7.2932-2940.1997

Induction of nitric oxide synthesis and xanthine oxidase and their roles in the antimicrobial mechanism against Salmonella typhimurium infection in mice.

K Umezawa 1, T Akaike 1, S Fujii 1, M Suga 1, K Setoguchi 1, A Ozawa 1, H Maeda 1
PMCID: PMC175411  PMID: 9199469

Abstract

The role of superoxide anion (O2-) and nitric oxide (NO) in the host defense mechanism against Salmonella typhimurium (LT-2) was examined by focusing on xanthine oxidase (XO) as an O2(-)-generating system and on inducible NO synthase (iNOS). When ICR mice were infected with a 0.1 50% lethal dose (2 x 10(5) CFU) of S. typhimurium, bacterial growth in the liver reached a peak value 3 days after infection (10(4.32) CFU/g of liver) and decreased thereafter. XO activity in the liver became maximum at 7 days after infection; the value was 34.6 +/- 1.4 mU/g of liver at 7 days (compared with 11.0 +/- 1.3 mU/g of liver before infection). The time profile of NO production in the liver as determined by electron spin resonance spectroscopy was consistent with that of XO activity. Histological examination of infected liver showed the formation of multiple microabscesses with granulomatous lesions consisting of polymorphonuclear cells and mononuclear cells, and iNOS-expressing cells were localized in the confined areas of the microabscesses. When XO inhibitors such as allopurinol and 4-amino-6-hydroxypyrazolo[3,4-d]pyrimidine (AHPP) were administered to the infected mice, the mortality of the mice was significantly increased (10 of 21 and 11 of 20 for the allopurinol- and AHPP-treated groups, respectively, versus 2 of 20 for control mice), and bacterial growth was significantly enhanced. A similar exacerbation of the infection was obtained with N(omega)-monomethyl-L-arginine (L-NMMA) treatment of the mice. Of considerable importance is that granuloma formation in the liver was poorly developed by treatment with either XO inhibitors or L-NMMA. These results suggest that XO and NO play an important role in the antimicrobial mechanism against S. typhimurium in mice.

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Selected References

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  1. Adamson G. M., Billings R. E. Cytokine toxicity and induction of NO synthase activity in cultured mouse hepatocytes. Toxicol Appl Pharmacol. 1993 Mar;119(1):100–107. doi: 10.1006/taap.1993.1048. [DOI] [PubMed] [Google Scholar]
  2. Akaike T., Ando M., Oda T., Doi T., Ijiri S., Araki S., Maeda H. Dependence on O2- generation by xanthine oxidase of pathogenesis of influenza virus infection in mice. J Clin Invest. 1990 Mar;85(3):739–745. doi: 10.1172/JCI114499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Akaike T., Noguchi Y., Ijiri S., Setoguchi K., Suga M., Zheng Y. M., Dietzschold B., Maeda H. Pathogenesis of influenza virus-induced pneumonia: involvement of both nitric oxide and oxygen radicals. Proc Natl Acad Sci U S A. 1996 Mar 19;93(6):2448–2453. doi: 10.1073/pnas.93.6.2448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Badwey J. A., Karnovsky M. L. Active oxygen species and the functions of phagocytic leukocytes. Annu Rev Biochem. 1980;49:695–726. doi: 10.1146/annurev.bi.49.070180.003403. [DOI] [PubMed] [Google Scholar]
  5. Beckman J. S., Beckman T. W., Chen J., Marshall P. A., Freeman B. A. Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1620–1624. doi: 10.1073/pnas.87.4.1620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Beckman J. S., Parks D. A., Pearson J. D., Marshall P. A., Freeman B. A. A sensitive fluorometric assay for measuring xanthine dehydrogenase and oxidase in tissues. Free Radic Biol Med. 1989;6(6):607–615. doi: 10.1016/0891-5849(89)90068-3. [DOI] [PubMed] [Google Scholar]
  7. Brunelli L., Crow J. P., Beckman J. S. The comparative toxicity of nitric oxide and peroxynitrite to Escherichia coli. Arch Biochem Biophys. 1995 Jan 10;316(1):327–334. doi: 10.1006/abbi.1995.1044. [DOI] [PubMed] [Google Scholar]
  8. Castro L., Rodriguez M., Radi R. Aconitase is readily inactivated by peroxynitrite, but not by its precursor, nitric oxide. J Biol Chem. 1994 Nov 25;269(47):29409–29415. [PubMed] [Google Scholar]
  9. Clancy R. M., Abramson S. B. Nitric oxide: a novel mediator of inflammation. Proc Soc Exp Biol Med. 1995 Nov;210(2):93–101. doi: 10.3181/00379727-210-43927aa. [DOI] [PubMed] [Google Scholar]
  10. Collins F. M., Mackaness G. B., Blanden R. V. Infection-immunity in experimental salmonellosis. J Exp Med. 1966 Oct 1;124(4):601–619. doi: 10.1084/jem.124.4.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Doi K., Akaike T., Horie H., Noguchi Y., Fujii S., Beppu T., Ogawa M., Maeda H. Excessive production of nitric oxide in rat solid tumor and its implication in rapid tumor growth. Cancer. 1996 Apr 15;77(8 Suppl):1598–1604. doi: 10.1002/(SICI)1097-0142(19960415)77:8<1598::AID-CNCR27>3.0.CO;2-U. [DOI] [PubMed] [Google Scholar]
  12. Doi T., Ando M., Akaike T., Suga M., Sato K., Maeda H. Resistance to nitric oxide in Mycobacterium avium complex and its implication in pathogenesis. Infect Immun. 1993 May;61(5):1980–1989. doi: 10.1128/iai.61.5.1980-1989.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dupont G. P., Huecksteadt T. P., Marshall B. C., Ryan U. S., Michael J. R., Hoidal J. R. Regulation of xanthine dehydrogenase and xanthine oxidase activity and gene expression in cultured rat pulmonary endothelial cells. J Clin Invest. 1992 Jan;89(1):197–202. doi: 10.1172/JCI115563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Falciani F., Ghezzi P., Terao M., Cazzaniga G., Garattini E. Interferons induce xanthine dehydrogenase gene expression in L929 cells. Biochem J. 1992 Aug 1;285(Pt 3):1001–1008. doi: 10.1042/bj2851001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Farr S. B., Kogoma T. Oxidative stress responses in Escherichia coli and Salmonella typhimurium. Microbiol Rev. 1991 Dec;55(4):561–585. doi: 10.1128/mr.55.4.561-585.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Granger D. L., Hibbs J. B., Jr, Perfect J. R., Durack D. T. Specific amino acid (L-arginine) requirement for the microbiostatic activity of murine macrophages. J Clin Invest. 1988 Apr;81(4):1129–1136. doi: 10.1172/JCI113427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Granger D. N., Kubes P. The microcirculation and inflammation: modulation of leukocyte-endothelial cell adhesion. J Leukoc Biol. 1994 May;55(5):662–675. [PubMed] [Google Scholar]
  18. Grum C. M., Gross T. J., Mody C. H., Sitrin R. G. Expression of xanthine oxidase activity by murine leukocytes. J Lab Clin Med. 1990 Aug;116(2):211–218. [PubMed] [Google Scholar]
  19. Hausladen A., Fridovich I. Superoxide and peroxynitrite inactivate aconitases, but nitric oxide does not. J Biol Chem. 1994 Nov 25;269(47):29405–29408. [PubMed] [Google Scholar]
  20. Huie R. E., Padmaja S. The reaction of no with superoxide. Free Radic Res Commun. 1993;18(4):195–199. doi: 10.3109/10715769309145868. [DOI] [PubMed] [Google Scholar]
  21. Ikeda T., Shimokata K., Daikoku T., Fukatsu T., Tsutsui Y., Nishiyama Y. Pathogenesis of cytomegalovirus-associated pneumonitis in ICR mice: possible involvement of superoxide radicals. Arch Virol. 1992;127(1-4):11–24. doi: 10.1007/BF01309571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. James S. L., Glaven J. Macrophage cytotoxicity against schistosomula of Schistosoma mansoni involves arginine-dependent production of reactive nitrogen intermediates. J Immunol. 1989 Dec 15;143(12):4208–4212. [PubMed] [Google Scholar]
  23. James S. L. Role of nitric oxide in parasitic infections. Microbiol Rev. 1995 Dec;59(4):533–547. doi: 10.1128/mr.59.4.533-547.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Jarasch E. D., Grund C., Bruder G., Heid H. W., Keenan T. W., Franke W. W. Localization of xanthine oxidase in mammary-gland epithelium and capillary endothelium. Cell. 1981 Jul;25(1):67–82. doi: 10.1016/0092-8674(81)90232-4. [DOI] [PubMed] [Google Scholar]
  25. Kooy N. W., Royall J. A., Ye Y. Z., Kelly D. R., Beckman J. S. Evidence for in vivo peroxynitrite production in human acute lung injury. Am J Respir Crit Care Med. 1995 Apr;151(4):1250–1254. doi: 10.1164/ajrccm/151.4.1250. [DOI] [PubMed] [Google Scholar]
  26. Kubes P. Nitric oxide affects microvascular permeability in the intact and inflamed vasculature. Microcirculation. 1995 Sep;2(3):235–244. doi: 10.3109/10739689509146769. [DOI] [PubMed] [Google Scholar]
  27. Liew F. Y., Millott S., Parkinson C., Palmer R. M., Moncada S. Macrophage killing of Leishmania parasite in vivo is mediated by nitric oxide from L-arginine. J Immunol. 1990 Jun 15;144(12):4794–4797. [PubMed] [Google Scholar]
  28. Lowenstein C. J., Snyder S. H. Nitric oxide, a novel biologic messenger. Cell. 1992 Sep 4;70(5):705–707. doi: 10.1016/0092-8674(92)90301-r. [DOI] [PubMed] [Google Scholar]
  29. Miles A. M., Bohle D. S., Glassbrenner P. A., Hansert B., Wink D. A., Grisham M. B. Modulation of superoxide-dependent oxidation and hydroxylation reactions by nitric oxide. J Biol Chem. 1996 Jan 5;271(1):40–47. doi: 10.1074/jbc.271.1.40. [DOI] [PubMed] [Google Scholar]
  30. Miyamoto Y., Akaike T., Yoshida M., Goto S., Horie H., Maeda H. Potentiation of nitric oxide-mediated vasorelaxation by xanthine oxidase inhibitors. Proc Soc Exp Biol Med. 1996 Apr;211(4):366–373. doi: 10.3181/00379727-211-43982. [DOI] [PubMed] [Google Scholar]
  31. Nathan C. Nitric oxide as a secretory product of mammalian cells. FASEB J. 1992 Sep;6(12):3051–3064. [PubMed] [Google Scholar]
  32. Ohshima H., Bartsch H. Chronic infections and inflammatory processes as cancer risk factors: possible role of nitric oxide in carcinogenesis. Mutat Res. 1994 Mar 1;305(2):253–264. doi: 10.1016/0027-5107(94)90245-3. [DOI] [PubMed] [Google Scholar]
  33. Palmer R. M., Ferrige A. G., Moncada S. Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature. 1987 Jun 11;327(6122):524–526. doi: 10.1038/327524a0. [DOI] [PubMed] [Google Scholar]
  34. Rinaldo J. E., Clark M., Parinello J., Shepherd V. L. Nitric oxide inactivates xanthine dehydrogenase and xanthine oxidase in interferon-gamma-stimulated macrophages. Am J Respir Cell Mol Biol. 1994 Nov;11(5):625–630. doi: 10.1165/ajrcmb.11.5.7524568. [DOI] [PubMed] [Google Scholar]
  35. Rubbo H., Darley-Usmar V., Freeman B. A. Nitric oxide regulation of tissue free radical injury. Chem Res Toxicol. 1996 Jul-Aug;9(5):809–820. doi: 10.1021/tx960037q. [DOI] [PubMed] [Google Scholar]
  36. Setoguchi K., Takeya M., Akaike T., Suga M., Hattori R., Maeda H., Ando M., Takahashi K. Expression of inducible nitric oxide synthase and its involvement in pulmonary granulomatous inflammation in rats. Am J Pathol. 1996 Dec;149(6):2005–2022. [PMC free article] [PubMed] [Google Scholar]
  37. Sheffler L. A., Wink D. A., Melillo G., Cox G. W. Exogenous nitric oxide regulates IFN-gamma plus lipopolysaccharide-induced nitric oxide synthase expression in mouse macrophages. J Immunol. 1995 Jul 15;155(2):886–894. [PubMed] [Google Scholar]
  38. Stuehr D. J., Griffith O. W. Mammalian nitric oxide synthases. Adv Enzymol Relat Areas Mol Biol. 1992;65:287–346. doi: 10.1002/9780470123119.ch8. [DOI] [PubMed] [Google Scholar]
  39. Suematsu M., Suzuki H., Ishii H., Kato S., Yanagisawa T., Asako H., Suzuki M., Tsuchiya M. Early midzonal oxidative stress preceding cell death in hypoperfused rat liver. Gastroenterology. 1992 Sep;103(3):994–1001. doi: 10.1016/0016-5085(92)90034-v. [DOI] [PubMed] [Google Scholar]
  40. Szabó C., Salzman A. L., Ischiropoulos H. Endotoxin triggers the expression of an inducible isoform of nitric oxide synthase and the formation of peroxynitrite in the rat aorta in vivo. FEBS Lett. 1995 Apr 24;363(3):235–238. doi: 10.1016/0014-5793(95)00322-z. [DOI] [PubMed] [Google Scholar]
  41. Sztein M. B., Wasserman S. S., Tacket C. O., Edelman R., Hone D., Lindberg A. A., Levine M. M. Cytokine production patterns and lymphoproliferative responses in volunteers orally immunized with attenuated vaccine strains of Salmonella typhi. J Infect Dis. 1994 Dec;170(6):1508–1517. doi: 10.1093/infdis/170.6.1508. [DOI] [PubMed] [Google Scholar]
  42. Takahashi M., Ushijima T., Ozaki Y. Changes in hepatic superoxide dismutase and xanthine oxidase activity in mice infected with Salmonella typhimurium and Pseudomonas aeruginosa. J Med Microbiol. 1988 Aug;26(4):281–284. doi: 10.1099/00222615-26-4-281. [DOI] [PubMed] [Google Scholar]
  43. Terao M., Cazzaniga G., Ghezzi P., Bianchi M., Falciani F., Perani P., Garattini E. Molecular cloning of a cDNA coding for mouse liver xanthine dehydrogenase. Regulation of its transcript by interferons in vivo. Biochem J. 1992 May 1;283(Pt 3):863–870. doi: 10.1042/bj2830863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Tubaro E., Lotti B., Cavallo G., Croce C., Borelli G. Liver xanthine oxidase increase in mice in three patholgoical models. A possible defence mechanism. Biochem Pharmacol. 1980 Jul 1;29(13):1939–1943. doi: 10.1016/0006-2952(80)90107-0. [DOI] [PubMed] [Google Scholar]
  45. Tubaro E., Lotti B., Santiangeli C., Cavallo G. Xanthine oxidase increase in polymorphonuclear leucocytes and macrophages in mice in three pathological situations. Biochem Pharmacol. 1980 Jul 1;29(13):1945–1948. doi: 10.1016/0006-2952(80)90108-2. [DOI] [PubMed] [Google Scholar]
  46. Umezawa K., Ohnishi N., Tanaka K., Kamiya S., Koga Y., Nakazawa H., Ozawa A. Granulation in livers of mice infected with Salmonella typhimurium is caused by superoxide released from host phagocytes. Infect Immun. 1995 Nov;63(11):4402–4408. doi: 10.1128/iai.63.11.4402-4408.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Uppu R. M., Squadrito G. L., Pryor W. A. Acceleration of peroxynitrite oxidations by carbon dioxide. Arch Biochem Biophys. 1996 Mar 15;327(2):335–343. doi: 10.1006/abbi.1996.0131. [DOI] [PubMed] [Google Scholar]
  48. Yoshida K., Akaike T., Doi T., Sato K., Ijiri S., Suga M., Ando M., Maeda H. Pronounced enhancement of .NO-dependent antimicrobial action by an .NO-oxidizing agent, imidazolineoxyl N-oxide. Infect Immun. 1993 Aug;61(8):3552–3555. doi: 10.1128/iai.61.8.3552-3555.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Yoshida M., Akaike T., Wada Y., Sato K., Ikeda K., Ueda S., Maeda H. Therapeutic effects of imidazolineoxyl N-oxide against endotoxin shock through its direct nitric oxide-scavenging activity. Biochem Biophys Res Commun. 1994 Jul 29;202(2):923–930. doi: 10.1006/bbrc.1994.2018. [DOI] [PubMed] [Google Scholar]

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