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Esophageal cancer-related gene-4 (ECRG4) interactions with the innate immunity receptor complex

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Abstract

Objective and design

The human c2orf40 gene encodes a tumor suppressor gene called esophageal cancer-related gene-4 (ECRG4) with pro- and anti-inflammatory activities that depend on cell surface processing. Here, we investigated its physical and functional association with the innate immunity receptor complex.

Methods

Interactions between ECRG4 and the innate immunity receptor complex were assessed by flow cytometry, immunohistochemistry, confocal microscopy, and co-immunoprecipitation. Phage display was used for ligand targeting to cells that overexpress the TLR4–MD2–CD14.

Results

Immunoprecipitation and immunohistochemical studies demonstrate a physical interaction between ECRG4 and TLR4–MD2–CD14 on human granulocytes. Flow cytometry shows ECRG4 on the cell surface of a subset of CD14+ and CD16+ leukocytes. In a cohort of trauma patients, the C-terminal 16 amino acid domain of ECRG4 (ECRG4133–148) appears to be processed and shed, presumably at a thrombin-like consensus sequence. Phage targeting this putative ligand shows that this peptide sequence internalizes into cells through the TLR4/CD14/MD2 complex, but modulates inflammation through non-canonical, NFκB signal transduction.

Conclusions

ECRG4 is present on the surface of human monocytes and granulocytes. Its interaction with the human innate immunity receptor complex supports a role for cell surface activation of ECRG4 during inflammation and implicates this receptor in its mechanism of action.

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Acknowledgments

The authors are indebted to Tran Nguyen, Emelie Amburn and Alexandra Ortiz-Borboa for expert technical assistance. Research was supported by a Mentored Young Investigator Award from the Hydrocephalus Association (S.P.), the National Institutes of Health through a P20 Exploratory Center grant for Wound Healing Research from the NIGMS (P20-GM078421) to A.B. and from grants EY018479 (A.B.), DK085871 (A.B.) and HL73396 (B.P.E.) and supplemental funding by the National Eye Institute (NEI) and NIGMS through the American Recovery Act (ARRA). Studies with human blood were funded by a grant from the American Burn Association (RC).

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Authors and Affiliations

  1. Division of Trauma, Surgical Critical Care, Burns, and Acute Care Surgery, Department of Surgery, University of California, San Diego, San Diego, CA, USA

    Sonia Podvin, Xitong Dang, Morgan Meads, Arwa Kurabi, Todd Costantini, Brian P. Eliceiri, Andrew Baird & Raul Coimbra

  2. Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, La Jolla, CA, 92093, USA

    Sonia Podvin

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  1. Sonia Podvin
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  2. Xitong Dang
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Correspondence to Andrew Baird.

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Responsible Editor: Liwu Li.

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Podvin, S., Dang, X., Meads, M. et al. Esophageal cancer-related gene-4 (ECRG4) interactions with the innate immunity receptor complex. Inflamm. Res. 64, 107–118 (2015). https://doi.org/10.1007/s00011-014-0789-2

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  • DOI: https://doi.org/10.1007/s00011-014-0789-2

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