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Spermine oxidase is a regulator of macrophage host response to Helicobacter pylori: enhancement of antimicrobial nitric oxide generation by depletion of spermine

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Abstract

The gastric pathogen Helicobacter pylori causes peptic ulcer disease and gastric cancer. We have reported that in H. pylori-activated macrophages, nitric oxide (NO) derived from inducible NO synthase (iNOS) can kill the bacterium, iNOS protein expression is dependent on uptake of its substrate l-arginine (l-Arg), the polyamine spermine can inhibit iNOS translation by inhibiting l-Arg uptake, and inhibition of polyamine synthesis enhances NO-mediated bacterial killing. Because spermine oxidase (SMO), which back-converts spermine to spermidine, is induced in macrophages by H. pylori, we determined its role in iNOS-dependent host defense. SMO shRNA knockdown in RAW 264.7 murine macrophages resulted in a marked decrease in H. pylori-stimulated iNOS protein, but not mRNA expression, and a 90 % reduction in NO levels; NO production was also inhibited in primary murine peritoneal macrophages with SMO knockdown. There was an increase in spermine levels after H. pylori stimulation that rapidly decreased, while SMO knockdown caused a greater increase in spermine that was sustained. With SMO knockdown, l-Arg uptake and killing of H. pylori by macrophages was prevented. The overexpression of SMO by transfection of an expression plasmid prevented the H. pylori-stimulated increase in spermine levels, and led to increased l-Arg uptake, iNOS protein expression and NO production, and H. pylori killing. In two human monocytic cell lines, U937 and THP-1, overexpression of SMO caused a significant enhancement of NO production with H. pylori stimulation. By depleting spermine, SMO can abrogate the inhibitory effect of polyamines on innate immune responses to H. pylori by enhancing antimicrobial NO production.

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Abbreviations

H. pylori :

Helicobacter pylori

NO:

Nitric oxide

iNOS:

Inducible NO synthase

l-Arg:

l-Arginine

Arg1:

Arginase I

Arg2:

Arginase II

ODC:

Ornithine decarboxylase

SMO:

Spermine oxidase

H2O2 :

Hydrogen peroxide

siRNA:

Small interfering RNA

shRNA:

Small hairpin RNA

DFMO:

Difluoromethylornithine

SSAT:

Spermidine/spermine N 1-acetyltransferase

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Acknowledgments

This work was supported by National Institutes of Health grants R01DK053620, R01AT004821, P01CA028842, and P01CA116087 (to K.T.W.), K01AT007324 (to R.C.), R01CA051085 and R01CA098454 (to R.A.C.), the Vanderbilt University Digestive Disease Research Center grant (P30DK058404), and a Merit Review Grant (1I01BX001453) from the Office of Medical Research, Department of Veterans Affairs (to K.T.W.).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Division of Gastroenterology, Department of Medicine, Vanderbilt University School of Medicine, 1030C MRBIV, 2215 Garland Avenue, Nashville, TN, 37232, USA

    Rupesh Chaturvedi, Mohammad Asim, Daniel P. Barry, Jeanetta W. Frye & Keith T. Wilson

  2. Department of Cancer Biology, Vanderbilt University School of Medicine, Nashville, TN, USA

    Keith T. Wilson

  3. Department of Pathology, Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, TN, USA

    Keith T. Wilson

  4. Veterans Affairs Tennessee Valley Healthcare System, Nashville, TN, USA

    Mohammad Asim, Daniel P. Barry & Keith T. Wilson

  5. The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA

    Robert A. Casero Jr.

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  1. Rupesh Chaturvedi
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  2. Mohammad Asim
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  3. Daniel P. Barry
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  5. Robert A. Casero Jr.
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  6. Keith T. Wilson
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Correspondence to Keith T. Wilson.

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Chaturvedi, R., Asim, M., Barry, D.P. et al. Spermine oxidase is a regulator of macrophage host response to Helicobacter pylori: enhancement of antimicrobial nitric oxide generation by depletion of spermine. Amino Acids 46, 531–542 (2014). https://doi.org/10.1007/s00726-013-1531-z

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  • DOI: https://doi.org/10.1007/s00726-013-1531-z

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