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The microbiota regulates neutrophil homeostasis and host resistance to Escherichia coli K1 sepsis in neonatal mice

Nature Medicine volume 20, pages 524–530 (2014)Cite this article

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Abstract

Neonatal colonization by microbes, which begins immediately after birth, is influenced by gestational age and the mother's microbiota and is modified by exposure to antibiotics1. In neonates, prolonged duration of antibiotic therapy is associated with increased risk of late-onset sepsis (LOS)2, a disorder controlled by neutrophils3. A role for the microbiota in regulating neutrophil development and susceptibility to sepsis in the neonate remains unclear. We exposed pregnant mouse dams to antibiotics in drinking water to limit transfer of maternal microbes to the neonates. Antibiotic exposure of dams decreased the total number and composition of microbes in the intestine of the neonates. This was associated with decreased numbers of circulating and bone marrow neutrophils and granulocyte/macrophage–restricted progenitor cells in the bone marrow of antibiotic-treated and germ-free neonates. Antibiotic exposure of dams reduced the number of interleukin-17 (IL-17)-producing cells in the intestine and production of granulocyte colony–stimulating factor (G-CSF). Granulocytopenia was associated with impaired host defense and increased susceptibility to Escherichia coli K1 and Klebsiella pneumoniae sepsis in antibiotic-treated neonates, which could be partially reversed by administration of G-CSF. Transfer of a normal microbiota into antibiotic-treated neonates induced IL-17 production by group 3 innate lymphoid cells (ILCs) in the intestine, increasing plasma G-CSF levels and neutrophil numbers in a Toll-like receptor 4 (TLR4)– and myeloid differentiation factor 88 (MyD88)–dependent manner and restored IL-17–dependent resistance to sepsis. Specific depletion of ILCs prevented IL-17– and G-CSF–dependent granulocytosis and resistance to sepsis. These data support a role for the intestinal microbiota in regulation of granulocytosis, neutrophil homeostasis and host resistance to sepsis in neonates.

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Figure 1: Perinatal antibiotic exposure alters the pattern of microbial colonization in the intestine and attenuates postnatal granulocytosis.
Figure 2: Microbiota regulates postnatal granulocytosis and controls host resistance to E. coli.
Figure 3: Microbiota-derived signals regulate postnatal granulocytosis via a IL-17– and G-CSF–dependent pathway.
Figure 4: Microbiota-derived signals regulate postnatal granulocytosis through a TLR4 and MyD88 dependent pathway.

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Acknowledgements

We thank D. Artis (Department of Microbiology and Pathology, University of Pennsylvania) for providing GF mice. We thank N. Butz for her assistance with microbial DNA isolation. We thank the Children's Hospital of Philadelphia Research Institute Flow Cytometry and Cell Sorting Core Laboratory for technical advice and support. We thank S. Guttentag, K. Hudock and C. Hergott for their helpful comments. H.S.D. is supported by 5T32HD060556, P.M.O. is supported by 5R01AI093566, J.K.K. is supported by 5R01HL062052, 3R37HL079142 and 5P60AA009803, J.N.W. is supported by 1R01AI105168 and 5R01AI038446 and G.S.W. is supported by 1R01AI099479 and 5R01HL105834.

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Authors and Affiliations

  1. Division of Neonatology, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA

    Hitesh S Deshmukh, Yuhong Liu, Ogechukwu R Menkiti, Junjie Mei, Ning Dai & G Scott Worthen

  2. Department of Pediatrics, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, USA

    Hitesh S Deshmukh, Ogechukwu R Menkiti, Jeffrey N Weiser & G Scott Worthen

  3. Department of Pathology and Laboratory Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, USA

    Claire E O'Leary & Paula M Oliver

  4. Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA

    Jay K Kolls

  5. Department of Microbiology, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, USA

    Jeffrey N Weiser

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  1. Hitesh S Deshmukh
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Contributions

H.S.D., J.N.W. and G.S.W. conceived of the study. H.S.D. and G.S.W. designed the experiments. P.M.O. and J.K.K. provided reagents. H.S.D., O.R.M., Y.L., N.D., J.M. and C.E.O. carried out experiments. H.S.D. and G.S.W. analyzed the data and wrote the manuscript.

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Correspondence to Hitesh S Deshmukh or G Scott Worthen.

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Deshmukh, H., Liu, Y., Menkiti, O. et al. The microbiota regulates neutrophil homeostasis and host resistance to Escherichia coli K1 sepsis in neonatal mice. Nat Med 20, 524–530 (2014). https://doi.org/10.1038/nm.3542

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