Journal of Clinical Immunology

, Volume 34, Issue 3, pp 331–339 | Cite as

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

  • Dhaarini Murugan
  • Michael H. Albert
  • Jörg Langemeier
  • Jens Bohne
  • Jacek Puchalka
  • Päivi M. Järvinen
  • Fabian Hauck
  • Anne K. Klenk
  • Christine Prell
  • Stephanie Schatz
  • Jana Diestelhorst
  • Barbara Sciskala
  • Naschla Kohistani
  • Bernd H. Belohradsky
  • Susanna Müller
  • Thomas Kirchner
  • Mark R. Walter
  • Philip Bufler
  • Aleixo M. Muise
  • Scott B. Snapper
  • Sibylle Koletzko
  • Christoph Klein
  • Daniel Kotlarz
Original Research

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.

Keywords

Interleukin-10 very early onset inflammatory bowel disease children hematopoietic stem cell transplantation 

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

References

  1. 1.
    Abraham C, Cho JH. Inflammatory bowel disease. N Engl J Med. 2009;361(21):2066–78.PubMedCentralPubMedCrossRefGoogle Scholar
  2. 2.
    Podolsky DK. Inflammatory bowel disease. N Engl J Med. 2002;347(6):417–29.PubMedCrossRefGoogle Scholar
  3. 3.
    Mizoguchi A, Mizoguchi E. Inflammatory bowel disease, past, present and future: lessons from animal models. J Gastroenterol. 2008;43(1):1–17.PubMedCrossRefGoogle Scholar
  4. 4.
    Cho JH. The genetics and immunopathogenesis of inflammatory bowel disease. Nat Rev Immunol. 2008;8(6):458–66.PubMedCrossRefGoogle Scholar
  5. 5.
    Wellcome Trust Case Control C. Genome-wide association study of 14,000 cases of seven common diseases and 3,000 shared controls. Nature. 2007;447(7145):661–78.CrossRefGoogle Scholar
  6. 6.
    Mathew CG. New links to the pathogenesis of Crohn disease provided by genome-wide association scans. Nat Rev Genet. 2008;9(1):9–14.PubMedCrossRefGoogle Scholar
  7. 7.
    Samuels ME, Majewski J, Alirezaie N, Fernandez I, Casals F, Patey N, et al. Exome sequencing identifies mutations in the gene TTC7A in French-Canadian cases with hereditary multiple intestinal atresia. J Med Gen. 2013;50(5):324–9.CrossRefGoogle Scholar
  8. 8.
    Chen R, Giliani S, Lanzi G, Mias GI, Lonardi S, Dobbs K, et al. Whole-exome sequencing identifies tetratricopeptide repeat domain 7A (TTC7A) mutations for combined immunodeficiency with intestinal atresias. J Allergy Clin Immunol. 2013.Google Scholar
  9. 9.
    Griffiths AM. Specificities of inflammatory bowel disease in childhood. Best Pract Res Clin Gastroenterol. 2004;18(3):509–23.PubMedCrossRefGoogle Scholar
  10. 10.
    Heyman MB, Kirschner BS, Gold BD, Ferry G, Baldassano R, Cohen SA, et al. Children with early-onset inflammatory bowel disease (IBD): analysis of a pediatric IBD consortium registry. J Pediatr. 2005;146(1):35–40.PubMedCrossRefGoogle Scholar
  11. 11.
    Ruemmele FM, El Khoury MG, Talbotec C, Maurage C, Mougenot JF, Schmitz J, et al. Characteristics of inflammatory bowel disease with onset during the first year of life. J Pediatr Gastroenterol Nutr. 2006;43(5):603–9.PubMedCrossRefGoogle Scholar
  12. 12.
    Catucci M, Castiello MC, Pala F, Bosticardo M, Villa A. Autoimmunity in Wiskott-Aldrich syndrome: an unsolved enigma. Front Immunol. 2012;3:209.PubMedCentralPubMedCrossRefGoogle Scholar
  13. 13.
    Caudy AA, Reddy ST, Chatila T, Atkinson JP, Verbsky JW. CD25 deficiency causes an immune dysregulation, polyendocrinopathy, enteropathy, X-linked-like syndrome, and defective IL-10 expression from CD4 lymphocytes. J Allergy Clin Immunol. 2007;119(2):482–7.PubMedCrossRefGoogle Scholar
  14. 14.
    Rohr J, Pannicke U, Doring M, Schmitt-Graeff A, Wiech E, Busch A, et al. Chronic inflammatory bowel disease as key manifestation of atypical ARTEMIS deficiency. J Clin Immunol. 2010;30(2):314–20.PubMedCrossRefGoogle Scholar
  15. 15.
    Schappi MG, Smith VV, Goldblatt D, Lindley KJ, Milla PJ. Colitis in chronic granulomatous disease. Arch Dis Child. 2001;84(2):147–51.PubMedCentralPubMedCrossRefGoogle Scholar
  16. 16.
    Worthey EA, Mayer AN, Syverson GD, Helbling D, Bonacci BB, Decker B, et al. Making a definitive diagnosis: successful clinical application of whole exome sequencing in a child with intractable inflammatory bowel disease. Genet Med. 2011;13(3):255–62.PubMedCrossRefGoogle Scholar
  17. 17.
    Kouklakis G, Efremidou EI, Papageorgiou MS, Pavlidou E, Manolas KJ, Liratzopoulos N. Complicated Crohn’s-like colitis, associated with Hermansky-Pudlak syndrome, treated with Infliximab: a case report and brief review of the literature. J Med Case Rep. 2007;1:176.PubMedCentralPubMedCrossRefGoogle Scholar
  18. 18.
    Cheng LE, Kanwar B, Tcheurekdjian H, Grenert JP, Muskat M, Heyman MB, et al. Persistent systemic inflammation and atypical enterocolitis in patients with NEMO syndrome. Clin Immunol. 2009;132(1):124–31.PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    Yamaguchi T, Ihara K, Matsumoto T, Tsutsumi Y, Nomura A, Ohga S, et al. Inflammatory bowel disease-like colitis in glycogen storage disease type 1b. Inflamm Bowel Dis. 2001;7(2):128–32.PubMedCrossRefGoogle Scholar
  20. 20.
    Begin P, Patey N, Mueller P, Rasquin A, Sirard A, Klein C, et al. Inflammatory bowel disease and T cell lymphopenia in G6PC3 deficiency. J Clin Immunol. 2013;33(3):520–5.PubMedCrossRefGoogle Scholar
  21. 21.
    Conley ME, Dobbs AK, Quintana AM, Bosompem A, Wang YD, Coustan-Smith E, et al. Agammaglobulinemia and absent B lineage cells in a patient lacking the p85alpha subunit of PI3K. J Exp Med. 2012;209(3):463–70.PubMedCentralPubMedCrossRefGoogle Scholar
  22. 22.
    Glocker EO, Kotlarz D, Boztug K, Gertz EM, Schaffer AA, Noyan F, et al. Inflammatory bowel disease and mutations affecting the interleukin-10 receptor. N Engl J Med. 2009;361(21):2033–45.PubMedCentralPubMedCrossRefGoogle Scholar
  23. 23.
    Glocker EO, Frede N, Perro M, Sebire N, Elawad M, Shah N, et al. Infant colitis–it’s in the genes. Lancet. 2010;376(9748):1272.PubMedCrossRefGoogle Scholar
  24. 24.
    Kotlarz D, Beier R, Murugan D, Diestelhorst J, Jensen O, Boztug K, et al. Loss of interleukin-10 signaling and infantile inflammatory bowel disease: implications for diagnosis and therapy. Gastroenterology. 2012;143(2):347–55.PubMedCrossRefGoogle Scholar
  25. 25.
    Begue B, Verdier J, Rieux-Laucat F, Goulet O, Morali A, Canioni D, et al. Defective IL10 signaling defining a subgroup of patients with inflammatory bowel disease. Am J Gastroenterol. 2011;106(8):1544–55.PubMedCrossRefGoogle Scholar
  26. 26.
    Mao H, Yang W, Lee PP, Ho MH, Yang J, Zeng S, et al. Exome sequencing identifies novel compound heterozygous mutations of IL-10 receptor 1 in neonatal-onset Crohn’s disease. Genes Immun. 2012;13(5):437–42.PubMedCrossRefGoogle Scholar
  27. 27.
    Moran CJ, Walters TD, Guo CH, Kugathasan S, Klein C, Turner D, et al. IL-10R polymorphisms are associated with very-early-onset ulcerative colitis. Inflamm Bowel Dis. 2013;19(1):115–23.PubMedCentralPubMedCrossRefGoogle Scholar
  28. 28.
    Engelhardt KR, Shah N, Faizura-Yeop I, Kocacik Uygun DF, Frede N, Muise AM, et al. Clinical outcome in IL-10- and IL-10 receptor-deficient patients with or without hematopoietic stem cell transplantation. J Allergy Clin Immunol. 2013;131(3):825–30.PubMedCrossRefGoogle Scholar
  29. 29.
    Kelley LA, Sternberg MJ. Protein structure prediction on the web: a case study using the Phyre server. Nat Protoc. 2009;4(3):363–71.PubMedCrossRefGoogle Scholar
  30. 30.
    DeLano WL. The PyMOL Molecular Graphics System DeLano Scientific, San Carlos, CA, USA. http://www.pymol.org. 2002.
  31. 31.
    Kotlarz D, Zietara N, Uzel G, Weidemann T, Braun CJ, Diestelhorst J, et al. Loss-of-function mutations in the IL-21 receptor gene cause a primary immunodeficiency syndrome. J Exp Med. 2013;210(3):433–43.PubMedCentralPubMedCrossRefGoogle Scholar
  32. 32.
    Schneider CA, Rasband WS, Eliceiri KW. NIH Image to imageJ: 25 years of image analysis. Nat Methods. 2012;9(7):671–5.PubMedCrossRefGoogle Scholar
  33. 33.
    Neven B, Mamessier E, Bruneau J, Kaltenbach S, Kotlarz D, Suarez F, et al. A Mendelian predisposition to B cell lymphoma caused by IL-10R deficiency. Blood 2013. Accepted.Google Scholar
  34. 34.
    Josephson K, Logsdon NJ, Walter MR. Crystal structure of the IL-10/IL-10R1 complex reveals a shared receptor binding site. Immunity. 2001;15(1):35–46.PubMedCrossRefGoogle Scholar
  35. 35.
    Krawczak M, Reiss J, Cooper DN. The mutational spectrum of single base-pair substitutions in mRNA splice junctions of human genes: causes and consequences. Hum Genet. 1992;90(1–2):41–54.PubMedGoogle Scholar
  36. 36.
    Rogaev EI, Grigorenko AP, Faskhutdinova G, Kittler EL, Moliaka YK. Genotype analysis identifies the cause of the “royal disease”. Science. 2009;326(5954):817.PubMedCrossRefGoogle Scholar
  37. 37.
    Thapar N, Lindley KJ, Kiparissi F, Elawad MA, Ashworth M, Veys P, et al. Treatment of intractable ulcerating enterocolitis of infancy by allogeneic bone marrow transplantation. Clin Gastroenterol Hepatol. 2008;6(2):248–50.PubMedCrossRefGoogle Scholar
  38. 38.
    Luznik L, O’Donnell PV, Fuchs EJ. Post-transplantation cyclophosphamide for tolerance induction in HLA-haploidentical bone marrow transplantation. Semin Oncol. 2012;39(6):683–93.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Dhaarini Murugan
    • 1
  • Michael H. Albert
    • 1
  • Jörg Langemeier
    • 2
  • Jens Bohne
    • 2
  • Jacek Puchalka
    • 1
  • Päivi M. Järvinen
    • 1
  • Fabian Hauck
    • 1
  • Anne K. Klenk
    • 1
  • Christine Prell
    • 1
  • Stephanie Schatz
    • 1
  • Jana Diestelhorst
    • 1
  • Barbara Sciskala
    • 1
  • Naschla Kohistani
    • 1
  • Bernd H. Belohradsky
    • 1
  • Susanna Müller
    • 3
  • Thomas Kirchner
    • 3
  • Mark R. Walter
    • 4
  • Philip Bufler
    • 1
  • Aleixo M. Muise
    • 5
    • 6
    • 7
  • Scott B. Snapper
    • 8
    • 9
  • Sibylle Koletzko
    • 1
  • Christoph Klein
    • 1
  • Daniel Kotlarz
    • 1
    • 10
  1. 1.Dr. von Hauner Children’s HospitalLudwig Maximilians UniversityMunichGermany
  2. 2.Cell and Virus Genetics GroupInstitute for Virology, Hannover Medical SchoolHannoverGermany
  3. 3.Institute of PathologyLudwig Maximilians UniversityMunichGermany
  4. 4.Department of MicrobiologyUniversity of Alabama at BirminghamBirminghamUSA
  5. 5.SickKids Inflammatory Bowel Disease Center and Cell Biology Program, Research InstituteThe Hospital for Sick ChildrenTorontoCanada
  6. 6.Division of Gastroenterology, Hepatology, and Nutrition, Department of PediatricsUniversity of Toronto, The Hospital for Sick ChildrenTorontoCanada
  7. 7.Institute of Medical ScienceUniversity of TorontoTorontoCanada
  8. 8.Division of Pediatric Gastroenterology, Hepatology, and Nutrition, Department of MedicineChildren’s Hospital BostonBostonUSA
  9. 9.Division of Gastroenterology and Hepatology, Brigham & Women’s Hospital, Department of MedicineHarvard Medical SchoolBostonUSA
  10. 10.Dr. von Hauner Children’s HospitalLudwig Maximilians UniversityMunichGermany

Personalised recommendations