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TLR4, IL10RA, and NOD2 mutation in paediatric Crohn’s disease patients: an association with Mycobacterium avium subspecies paratuberculosis and TLR4 and IL10RA expression

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Abstract

Mycobacterium avium subspecies paratuberculosis (MAP) has been implicated in the pathogenesis of Crohn’s disease (CD). The role of CD susceptibility genes in association with these microbes is not known. Sixty-two early onset paediatric CD patients and 46 controls with known MAP status were analysed for an association with 34 single nucleotide polymorphisms (SNPs) from 18 CD susceptibility genes. Functional studies on peripheral blood mononuclear cells (PBMCs) were conducted on 17 CD patients with known CD mutations to assess IL-6, IL-10, and TNF-α expression upon stimulation with MAP precipitated protein derivative (PPD) and lipopolysaccharide (LPS). In addition, surface expression of IL10R and TLR4 on resting B cells, NK cells, T cells, and monocytes was assessed. A mutation in TLR4 (rs4986790) and IL10RA (rs22291130) was significantly associated with MAP-positive CD patients compared to MAP-negative CD patients (27.6 vs. 6.1 %, p = 0.021, and 62.1 vs. 33.3 %, p = 0.024, respectively). PPD and LPS significantly increased IL-6, IL-10, and TNF-α production in PBMCs. IL-10 and TNF-α production were significantly lower in a subgroup of CD patients (5/12) with a known NOD2 mutation. Receptor for IL-10 was significantly higher expressed on NK cells (CD56low) and on NK T cells harbouring a NOD2 mutations compared to wildtype cells (p = 0.031 and 0.005, respectively). TLR4 was significantly higher expressed on NK cells (CD56high) harbouring a NOD2 mutations compared to wildtype cells (p = 0.038).

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Acknowledgments

We thank the children and their families for their participation in this study. This work was supported by research grants from the Murdoch Childrens Research Institute, The CASS Foundation, and The Lynne Quayle Charitable Trust, Equity Trustees LTD. Dr. C. Kirkwood is supported by an NHMRC CDA Fellowship (609347). This work was supported by the Victorian Government Operational Infrastructure Support Program.

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Authors and Affiliations

  1. Enteric Virus Group, Murdoch Childrens Research Institute, Royal Children’s Hospital, Flemington Road, Parkville, VIC, 3052, Australia

    Josef Wagner & Carl D. Kirkwood

  2. Monash Centre for Inflammatory Diseases, Monash University, Clayton, VIC, 3168, Australia

    Narelle A. Skinner & Kumar Visvanathan

  3. Department of Pediatrics, University of Melbourne, Parkville, VIC, 3052, Australia

    Josef Wagner, Anthony G. Catto-Smith, Donald J. S. Cameron & Carl D. Kirkwood

  4. Department of Gastroenterology and Clinical Nutrition, Royal Children’s Hospital, Parkville, VIC, 3052, Australia

    Anthony G. Catto-Smith & Donald J. S. Cameron

  5. Australian Animal Health Laboratory, CSIRO Animal, Food and Health Sciences, East Geelong, VIC, 3219, Australia

    Wojtek P. Michalski

  6. Department of Microbiology, Latrobe University, Bundoora, VIC, 3086, Australia

    Carl D. Kirkwood

  7. Department of Medicine, St Vincents Hospital, University of Melbourne, Melbourne, Australia

    Kumar Visvanathan

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  1. Josef Wagner
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Correspondence to Josef Wagner.

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Wagner, J., Skinner, N.A., Catto-Smith, A.G. et al. TLR4, IL10RA, and NOD2 mutation in paediatric Crohn’s disease patients: an association with Mycobacterium avium subspecies paratuberculosis and TLR4 and IL10RA expression. Med Microbiol Immunol 202, 267–276 (2013). https://doi.org/10.1007/s00430-013-0290-5

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