Article | Published:

Infectious diseases and host defense

Leptin receptor q223r polymorphism influences neutrophil mobilization after Clostridium difficile infection

Mucosal Immunology volume 11, pages 947957 (2018) | Download Citation

Abstract

Clostridium difficile is the leading cause of nosocomial infections in the United States. Clinical disease outcomes after C. difficile infection (CDI) are dependent on intensity of host inflammatory responses. Specifically, peak peripheral white blood cell (WBC) count >20 × 109 l−1 is an indicator of adverse outcomes in CDI patients, and is associated with higher 30-day mortality. We show that homozygosity for a common single nucleotide polymorphism (Q to R mutation in leptin receptor that is present in up to 50% of people), significantly increases the risk of having peak peripheral WBC count >20 × 109 l−1 (odds ratio=5.41; P=0.0023) in CDI patients. In a murine model of CDI, we demonstrate that mice homozygous for the same single nucleotide polymorphism (RR mice) have more blood and tissue leukocytes (specifically neutrophils), exaggerated tissue inflammation, and higher mortality as compared with control mice, despite similar pathogen burden. Further, we show that neutrophilia in RR mice is mediated by gut microbiota-directed expression of CXC chemokine receptor 2 (CXCR2), which promotes the release of neutrophils from bone marrow reservoir. Overall these studies provide novel mechanistic insights into the role of human genetic polymorphisms and gut microbiota in regulating the fundamental biological process of CDI-induced neutrophilia.

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Acknowledgements

Human samples and data were collected in the laboratory of Dr. William A. Petri Jr. (University of Virginia) and financially supported by R01 AI-124214 to WAP. We thank TechLab, Inc. for generously providing TOX A/B ELISA kits. We would like to thank George S. Deepe Jr. (UC) and Senad Divanovic and Marie-Dominique Filippi (both at CCHMC) for critical review of the manuscript and valuable discussions; and Roman Jandarov (UC) for performing statistical analysis of patient data. We also thank the Live Microscopy Core in the department of Molecular and Cellular Physiology at UC and Research Flow Cytometry Core in the Division of Rheumatology at CCHMC for their assistance. This work was supported by the National Institutes of Health (NIH) K08-AI108801-01 to Rajat Madan. Microbiome analysis was supported by the NIH (T32ES010957-14) and the Centers for Disease Control (CDC) AR funding through a Broad Agency Announcement (200-2016-91939). This project was supported in part by PHS Grant P30 DK078392 (Research Pathology and Research Flow Cytometry Core) of the Digestive Disease Research Core Center in Cincinnati.

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Affiliations

  1. Division of Infectious Diseases, Department of Internal Medicine, University of Cincinnati, Cincinnati, Ohio, USA

    • S Jose
    • , A Mukherjee
    • , J Xue
    •  & R Madan
  2. Division of Infectious Diseases and International Health, Department of Medicine, University of Virginia, Charlottesville, Virginia, USA

    • M M Abhyankar
  3. Department of Infectious Diseases, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA

    • H Andersen
    •  & D B Haslam
  4. Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children’s Hospital Medical Center, Cincinnati, Ohio, USA

    • R Madan

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Competing interests

The authors declare no conflict of interest.

Corresponding author

Correspondence to R Madan.

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DOI

https://doi.org/10.1038/mi.2017.119

Author contributions

R.M. conceived the study and obtained funding. S.J. and R.M. designed the experiments and analyzed the data. S.J. conducted mouse studies. M.M.A. and R.M. collected and analyzed human data. A.M. and J.X. maintained the mouse colonies and assisted in experiments. H.A. and D.B.H. performed the shotgun sequencing and microbiota analysis. S.J. and R.M. wrote the manuscript.

SUPPLEMENTARY MATERIAL is linked to the online version of the paper at http://www.nature.com/mi