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Arginase 2 deletion leads to enhanced M1 macrophage activation and upregulated polyamine metabolism in response to Helicobacter pylori infection

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Abstract

We reported that arginase 2 (ARG2) deletion results in increased gastritis and decreased bacterial burden during Helicobacter pylori infection in mice. Our studies implicated a potential role for inducible nitric oxide (NO) synthase (NOS2), as Arg2 −/− mice exhibited increased NOS2 levels in gastric macrophages, and NO can kill H. pylori. We now bred Arg2 −/− to Nos2 −/− mice, and infected them with H. pylori. Compared to wild-type mice, both Arg2 −/− and Arg2 −/− ;Nos2 −/− mice exhibited increased gastritis and decreased colonization, the latter indicating that the effect of ARG2 deletion on bacterial burden was not mediated by NO. While Arg2 −/− mice demonstrated enhanced M1 macrophage activation, Nos2 −/− and Arg2 −/− ;Nos2 −/− mice did not demonstrate these changes, but exhibited increased CXCL1 and CXCL2 responses. There was an increased expression of the Th1/Th17 cytokines, interferon gamma and interleukin 17, in gastric tissues and splenic T-cells from Arg2 −/−, but not Nos2 −/− or Arg2 −/− ;Nos2 −/− mice. Gastric tissues from infected Arg2 −/− mice demonstrated increased expression of arginase 1, ornithine decarboxylase, adenosylmethionine decarboxylase 1, spermidine/spermine N 1-acetyltransferase 1, and spermine oxidase, along with increased spermine levels. These data indicate that ARG2 deletion results in compensatory upregulation of gastric polyamine synthesis and catabolism during H. pylori infection, which may contribute to increased gastric inflammation and associated decreased bacterial load. Overall, the finding of this study is that ARG2 contributes to the immune evasion of H. pylori by restricting M1 macrophage activation and polyamine metabolism.

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Abbreviations

AMD1:

Adenosylmethionine decarboxylase 1

ARG1:

Arginase 1; arginase, type I

ARG2:

Arginase 2; arginase, type II

CCL:

C–C chemokine ligand

CD:

Cluster of differentiation

CHIA1:

Chitinase, acidic 1

CXCL:

C–X–C chemokine ligand

FPL:

French-pressed lysate

IFN-γ:

Interferon gamma

IL:

Interleukin

M-CSF:

Macrophage colony stimulating factor

MOI:

Multiplicity of infection

NO:

Nitric oxide

NOS2:

Nitric oxide synthase 2; inducible nitric oxide synthase

ODC:

Ornithine decarboxylase

PMSS1:

Pre-mouse Sydney Strain 1

RT-PCR:

Real-time polymerase chain reaction

RETNLA:

Resistin like molecule alpha

SAT1:

Spermidine/spermine N 1-acetyltransferase 1

SMOX:

Spermine oxidase

SS1:

Sydney Strain 1

TGF:

Transforming growth factor

Th:

T helper

TNF:

Tumor necrosis factor

TNFSF14:

Tumor necrosis factor superfamily member 14

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Acknowledgments

We thank Alain P. Gobert (Vanderbilt University) for intellectual input and for his review of the manuscript. We thank Daniel P. Barry (Vanderbilt University) for his assistance with the breeding and maintenance of the mouse colonies utilized in this study. We thank Margaret Allaman (Vanderbilt University) for her assistance with the Luminex assays.

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Authors and Affiliations

  1. Department of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Vanderbilt University School of Medicine, 2215 Garland Avenue, 1030C Medical Research Building IV, Nashville, TN, 37232, USA

    Dana M. Hardbower & Keith T. Wilson

  2. Division of Gastroenterology, Hepatology and Nutrition, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, USA

    Dana M. Hardbower, Mohammad Asim, Thomas Verriere, Nuruddeen D. Lewis, Rupesh Chaturvedi, M. Blanca Piazuelo & Keith T. Wilson

  3. The Sydney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Tracy Murray-Stewart & Robert A. Casero Jr.

  4. School of Biotechnology, Jawaharlal Nehru University, New Delhi, India

    Rupesh Chaturvedi

  5. Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, TN, USA

    Keith T. Wilson

  6. Center for Mucosal Inflammation and Cancer, Vanderbilt University Medical Center, Nashville, TN, USA

    Keith T. Wilson

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

    Keith T. Wilson

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  1. Dana M. Hardbower
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  2. Mohammad Asim
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  3. Tracy Murray-Stewart
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  8. M. Blanca Piazuelo
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  9. Keith T. Wilson
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Correspondence to Keith T. Wilson.

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Funding

This work was funded by NIH Grants R01DK053620, R01AT004821, R01CA190612, P01CA116087, and P01CA028842 (K.T.W.), Merit Review Grant I01BX001453 from the United States Department of Veterans Affairs Biomedical Laboratory R&D (BLRD) Service (K.T.W.), the Vanderbilt Digestive Disease Research Center, supported by NIH Grant P30DK058404, the Thomas F. Frist Sr. Endowment (K.T.W.), and the Vanderbilt Center for Mucosal Inflammation and Cancer (K.T.W.). D.M.H. was supported by NIH Grants T32GM008554 and F31DK10715.

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The authors declare that no conflict of interest exists.

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Hardbower, D.M., Asim, M., Murray-Stewart, T. et al. Arginase 2 deletion leads to enhanced M1 macrophage activation and upregulated polyamine metabolism in response to Helicobacter pylori infection. Amino Acids 48, 2375–2388 (2016). https://doi.org/10.1007/s00726-016-2231-2

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