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Very Early Onset Inflammatory Bowel Disease Associated with Aberrant Trafficking of IL-10R1 and Cure by T Cell Replete Haploidentical Bone Marrow Transplantation

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Abstract

Purpose

Loss-of-function mutations in IL10 and IL10R cause very early onset inflammatory bowel disease (VEO-IBD). Here, we investigated the molecular pathomechanism of a novel intronic IL10RA mutation and describe a new therapeutic approach of T cell replete haploidentical hematopoietic stem cell transplantation (HSCT).

Methods

Clinical data were collected by chart review. Genotypes of IL10 and IL10R genes were determined by Sanger sequencing. Expression and function of mutated IL-10R1 were assessed by quantitative PCR, Western blot analysis, enzyme-linked immunosorbent assays, confocal microscopy, and flow cytometry.

Results

We identified a novel homozygous point mutation in intron 3 of the IL10RA (c.368-10C > G) in three related children with VEO-IBD. Bioinformatical analysis predicted an additional 3′ splice site created by the mutation. Quantitative PCR analysis showed normal mRNA expression of mutated IL10RA. Sequencing of the patient’s cDNA revealed an insertion of the last nine nucleotides of intron 3 as a result of aberrant splicing. Structure-based modeling suggested misfolding of mutated IL-10R1. Western blot analysis demonstrated a different N-linked glycosylation pattern of mutated protein. Immunofluorescence and FACS analysis revealed impaired expression of mutated IL-10R1 at the plasma membrane. In the absence of HLA-identical donors, T cell replete haploidentical HSCT was successfully performed in two patients.

Conclusions

Our findings expand the spectrum of IL10R mutations in VEO-IBD and emphasize the need for genetic diagnosis of mutations in conserved non-coding sequences of candidate genes. Transplantation of haploidentical stem cells represents a curative therapy in IL-10R-deficient patients, but may be complicated by non-engraftment.

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Abbreviations

ELISA:

enzyme-linked immunosorbent assay

IL-10R1:

Interleukin-10 receptor 1

LPS:

lipopolysaccharide

PBMCs:

peripheral blood mononuclear cells

TNF-α:

tumor necrosis factor-α

VEO-IBD:

very early onset inflammatory bowel disease

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Acknowledgments

We thank the patients, the interdisciplinary medical team, and clinicians involved in the care of these patients. The study is supported by grants from DFG (Gottfried-Wilhelm-Leibniz Program), BioSysNet, SFB1054, the Helmsley Charitable Trust, the Care-for-Rare Foundation (fellowship to D.K. and D.M.), and the DFG grant BO 2512/6-1 to J.B..

D.M. is a student of the PhD program Molecular Medicine at Hannover Biomedical Research School, Germany.

Conflict of Interest

The authors declare no competing financial interests.

Author information

Authors and Affiliations

  1. Dr. von Hauner Children’s Hospital, Ludwig Maximilians University, Munich, Germany

    Dhaarini Murugan, Michael H. Albert, Jacek Puchalka, Päivi M. Järvinen, Fabian Hauck, Anne K. Klenk, Christine Prell, Stephanie Schatz, Jana Diestelhorst, Barbara Sciskala, Naschla Kohistani, Bernd H. Belohradsky, Philip Bufler, Sibylle Koletzko, Christoph Klein & Daniel Kotlarz

  2. Cell and Virus Genetics Group, Institute for Virology, Hannover Medical School, Hannover, Germany

    Jörg Langemeier & Jens Bohne

  3. Institute of Pathology, Ludwig Maximilians University, Munich, Germany

    Susanna Müller & Thomas Kirchner

  4. Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL, USA

    Mark R. Walter

  5. SickKids Inflammatory Bowel Disease Center and Cell Biology Program, Research Institute, The Hospital for Sick Children, Toronto, Canada

    Aleixo M. Muise

  6. Division of Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, University of Toronto, The Hospital for Sick Children, Toronto, Canada

    Aleixo M. Muise

  7. Institute of Medical Science, University of Toronto, Toronto, Canada

    Aleixo M. Muise

  8. Division of Pediatric Gastroenterology, Hepatology, and Nutrition, Department of Medicine, Children’s Hospital Boston, Boston, USA

    Scott B. Snapper

  9. Division of Gastroenterology and Hepatology, Brigham & Women’s Hospital, Department of Medicine, Harvard Medical School, Boston, USA

    Scott B. Snapper

  10. Dr. von Hauner Children’s Hospital, Ludwig Maximilians University, Lindwurmstr. 4, 80337, Munich, Germany

    Daniel Kotlarz

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  1. Dhaarini Murugan
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  2. Michael H. Albert
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  21. Sibylle Koletzko
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Corresponding author

Correspondence to Daniel Kotlarz.

Additional information

Dhaarini Murugan and Michael H. Albert contributed equally to this work and should be considered aequo loco.

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Murugan, D., Albert, M.H., Langemeier, J. et al. Very Early Onset Inflammatory Bowel Disease Associated with Aberrant Trafficking of IL-10R1 and Cure by T Cell Replete Haploidentical Bone Marrow Transplantation. J Clin Immunol 34, 331–339 (2014). https://doi.org/10.1007/s10875-014-9992-8

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  • DOI: https://doi.org/10.1007/s10875-014-9992-8

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