Journal of Clinical Immunology

, Volume 37, Issue 8, pp 801–810 | Cite as

Functional Evaluation of an IKBKG Variant Suspected to Cause Immunodeficiency Without Ectodermal Dysplasia

  • Glynis Frans
  • Jutte van der Werff Ten Bosch
  • Leen Moens
  • Rik Gijsbers
  • Majid Changi-Ashtiani
  • Hassan Rokni-Zadeh
  • Mohammad Shahrooei
  • Greet Wuyts
  • Isabelle Meyts
  • Xavier BossuytEmail author
Original Article


Hypomorphic IKBKG mutations in males are typically associated with anhidrotic ectodermal dysplasia with immunodeficiency (EDA-ID). Some mutations cause immunodeficiency without EDA (NEMO-ID). The immunological profile associated with these NEMO-ID variants is not fully documented. We present a 2-year-old patient with suspected immunodeficiency in which a hemizygous p.Glu57Lys IKBKG variant was identified. At the age of 1 year, he had an episode of otitis media that evolved into a bilateral mastoiditis (Pseudomonas spp). Hypogammaglobulinemia, specific (polysaccharide) antibody deficiency, and low switched memory B cell subsets were noticed. The mother was heterozygous for the variant but had no signs of incontinentia pigmenti. Patient peripheral blood mononuclear cells produced low amounts of IL-6 after stimulation with IL-1β, Pam3CSK4, and FSL-1. In patient fibroblasts, IκB-α was degraded normally upon stimulation with IL-1β or TNF-α. Transduction of wild-type and variant NEMO in NEMO−/− deficient SV40 fibroblasts revealed a slight but significant reduction of IL-6 production upon stimulation with IL-1β and TNF-α. In conclusion, we demonstrated that p.Glu57Lys leads to specific immunological defects in vitro. No other pathogenic PID variants were identified through whole exome sequencing. As rare polymorphisms have been described in IKBKG and polygenic inheritance remains an option in the presented case, this study emphasizes the need for thorough functional and genetic evaluation when encountering and interpreting suspected disease-causing NEMO-ID variants.


Immunodeficiency NF-κB essential modulator NEMO toll-like receptors NF-κB pathway 



We thank Jean-Laurent Casanova for providing NEMO-deficient SV40 transformed fibroblasts. We are indebted to the patient and his parents.

Authorship Contributions

GF drafted the manuscript and conducted experiments. JVDWTB characterized the immune deficiency and coordinates the clinical care of the patient. LM, RG, and GW conducted experiments. HRZ, MC-A, and MS performed whole exome sequencing and bioinformatics analysis. XB supervised the routine laboratory immunology work up and TLR testing. IM and XB designed the study and finalized the manuscript. Each author has critically revised the final version of the manuscript and has read and approved the final manuscript.

Funding Information

This work was supported by a GOA grant from the Research Council of the KU Leuven, Belgium. IM is supported by a KOF mandate of the KU Leuven, Belgium.

Compliance with Ethical Standards

Conflicts of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

The study was performed in accordance with the 1964 Helsinki declaration and its later amendments. Informed consent was obtained for genetic analysis and report of the case. The study was approved by the Ethics Committee of UZ Leuven.

Supplementary material

10875_2017_448_MOESM1_ESM.docx (21 kb)
ESM 1 (DOCX 20 kb)


  1. 1.
    Karin M, Ben-Neriah Y. Phosphorylation meets ubiquitination: the control of NF-κB activity. Annu Rev Immunol. 2000;18:621–63.CrossRefPubMedGoogle Scholar
  2. 2.
    Hayden MS, Ghosh S. Shared principles in NF-κB signaling. Cell. 2008;132:344–62.CrossRefPubMedGoogle Scholar
  3. 3.
    Scheidereit C. IκB kinase complexes: gateways to NF-κB activation and transcription. Oncogene. 2006;25:6685–705.CrossRefPubMedGoogle Scholar
  4. 4.
    Fusco F, Bardaro T, Fimiani G, Mercadante V, Miano MG, Falco G, et al. Molecular analysis of the genetic defect in a large cohort of IP patients and identification of novel NEMO mutations interfering with NF-kappaB activation. Hum Mol Genet. 2004;13:1763–73.CrossRefPubMedGoogle Scholar
  5. 5.
    Uzel G. The range of defects associated with nuclear factor kappaB essential modulator. Curr Opin Allergy Clin Immunol. 2005;5:513–8.CrossRefPubMedGoogle Scholar
  6. 6.
    Courtois G, Gilmore TD. Mutations in the NF-κB signaling pathway: implications for human disease. Oncogene. 2006;25:6831–43.CrossRefPubMedGoogle Scholar
  7. 7.
    Hanson EP, Monaco-Shawver L, Solt LA, Madge LA, Banerjee PP, May MJ, et al. Hypomorphic nuclear factor-kappaB essential modulator mutation database and reconstitution system identifies phenotypic and immunologic diversity. J Allergy Clin Immunol. 2008;122:1169–77.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Fusco F, Pescatore A, Conte MI, Mirabelli P, Paciolla M, Esposito E, et al. EDA-ID and IP, two faces of the same coin: how the same IKBKG/NEMO mutation affecting the NF-κB pathway can cause immunodeficiency and/or inflammation. Int Rev Immunol. 2015;34:445–59.CrossRefPubMedGoogle Scholar
  9. 9.
    Boisson B, Puel A, Picard C, Casanova JL. Human IκBα gain of function: a severe and syndromic immunodeficiency. J Clin Immunol. 2017;37:397–412.CrossRefPubMedGoogle Scholar
  10. 10.
    Frans G, Meyts I, Picard C, Puel A, Zhang SY, Moens L, et al. Addressing diagnostic challenges in primary immunodeficiencies: laboratory evaluation of Toll-like receptor- and NF-κB-mediated immune responses. Crit Rev Clin Lab Sci. 2014;51:112–23.CrossRefPubMedGoogle Scholar
  11. 11.
    Rosenzweig SD, Holland SM. Defects in the interferon-gamma and interleukin-12 pathways. Immunol Rev. 2005;203:38–47.CrossRefPubMedGoogle Scholar
  12. 12.
    Filipe-Santos O, Bustamante J, Haverkamp MH, Vinolo E, Ku CL, Puel A, et al. X-linked susceptibility to mycobacteria is caused by mutations in NEMO impairing CD40-dependent IL-12 production. J Exp Med. 2006;203:1745–59.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Puel A, Reichenbach J, Bustamante J, Ku CL, Feinberg J, Döffinger R, et al. The NEMO mutation creating the most-upstream premature stop codon is hypomorphic because of a reinitation of translation. Am J Hum Genet. 2006;78:691–701.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Niehues T, Reichenbach J, Neubert J, Gudowius S, Puel A, Horneff G, et al. Nuclear factor kappaB essential modulator-deficient child with immunodeficiency yet without anhidrotic ectodermal dysplasia. J Allergy Clin Immunol. 2004;114:1456–62.CrossRefPubMedGoogle Scholar
  15. 15.
    Ku CL, Dupuis-Girod S, Dittrich AM, Bustamante J, Santos OF, Schulze I, et al. NEMO mutations in 2 unrelated boys with severe infections and conical teeth. Pediatrics. 2005;115:615–9.CrossRefGoogle Scholar
  16. 16.
    Salt BH, Niemela JE, Pandey R, Hanson EP, Deering RP, Quinones R, et al. IKBKG (nuclear factor-κB essential modulator) mutation can be associated with opportunistic infection without impairing Toll-like receptor function. J Allergy Clin Immunol. 2008;121:976–82.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Orange JS, Levy O, Brodeur SR, Krzewski K, Roy RM, Niemela JE, et al. Human nuclear factor kappa B essential modulator mutation can result in immunodeficiency without ectodermal dysplasia. J Allergy Clin Immunol. 2004;114:650–6.CrossRefPubMedGoogle Scholar
  18. 18.
    Orange JS, Jain A, Ballas ZK, Schneider LC, Geha RS, Bonilla FA. The presentation and natural history of immunodeficiency caused by nuclear factor kappaB essential modulator mutation. J Allergy Clin Immunol. 2004;113:725–33.CrossRefPubMedGoogle Scholar
  19. 19.
    Dai YS, Liang MG, Gellis SE, Bonilla FA, Schneider LC, Geha RS, et al. Characteristics of mycobacterial infection in patients with immunodeficiency and nuclear factor-kappaB essential modulator mutation, with or without ectodermal dysplasia. J Am Acad Dermatol. 2004;51:718–22.CrossRefPubMedGoogle Scholar
  20. 20.
    Tuerlinckx D, Vermeulen F, Pékus V, de Bilderling G, Glupczynski Y, Collet S, et al. Optimal assessment of the ability of children with recurrent respiratory tract infections to produce anti-polysaccharide antibodies. Clin Exp Immunol. 2007;149:295–302.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Shirkani A, Shahrooei M, Azizi G, Rokni-Zadeh H, Abolhassani H, Farrokhi S, et al. Novel mutation of ZAP-70-related combined immunodeficiency: first case from the National Iranian Registry and Review of the Literature. Immunol Investig. 2017;46:70–9.CrossRefGoogle Scholar
  22. 22.
    Li H, Durbin R. Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics. 2009;25:1754–60.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, Homer N, et al. The sequence alignment/map (SAM) format and SAMtools. Bioinformatics. 2009;25:2078–9.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Smahi A, Courtois G, Rabia SH, Doffinger R, Bodemer C, Munnich A, et al. The NF-kappaB signalling pathway in human diseases: from incontinentia pigmenti to ectodermal dysplasias and immune-deficiency syndromes. Hum Mol Genet. 2002;11:2371–5.CrossRefPubMedGoogle Scholar
  25. 25.
    Ibrahimi A, Vande Velde G, Reumers V, Toelen J, Thiry I, Vandeputte C, et al. Highly efficient multicistronic lentiviral vectors with peptide 2A sequences. Hum Gene Ther. 2009;20:845–60.CrossRefPubMedGoogle Scholar
  26. 26.
    Devora GA, Sun L, Chen Z, van Oers NS, Hanson EP, Orange JS, et al. A novel missense mutation in the nuclear factor-κB essential modulator (NEMO) gene resulting in impaired activation of the NF-κB pathway and a unique clinical phenotype presenting as MRSA subdural empyema. J Clin Immunol. 2010;30:881–5.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Keller MD, Petersen M, Ong P, Church J, Risma K, Burham J, et al. Hypohidrotic ectodermal dysplasia and immunodeficiency with coincident NEMO and EDA mutations. Front Immunol. 2011;2:61.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Tarpey PS, Smith R, Pleasance E, Whibley A, Edkins S, Hardy C, et al. A systematic, large-scale resequencing screen of X-chromosome coding exons in mental retardation. Nat Genet. 2009;41:535–43.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Rameix-Welti MA, Régnier CH, Bienaimé F, Blouin J, Schifferli J, Fridman WH, et al. Hereditary complement C7 deficiency in nine families: subtotal C7 deficiency revisited. Eur J Immunol. 2007;37:1377–85.CrossRefPubMedGoogle Scholar
  30. 30.
    Becker-Heck A, Zohn IE, Okabe N, Pollock A, Lenhart KB, Sullivan-Brown J, et al. The coiled-coil domain containing protein CCDC40 is essential for motile cilia function and left-right axis formation. Nat Genet. 2011;43:79–84.CrossRefPubMedGoogle Scholar
  31. 31.
    Exome Aggregation Consortium (ExAC), Cambridge, MA, USA. Website: Accessed on: 5 Aug 2016.
  32. 32.
    Castigli E, Wilson SA, Garibyan L, Rachid R, Bonilla F, Schneider L, et al. TACI is mutant in common variable immunodeficiency and IgA deficiency. Nat Genet. 2005;37:829–34.CrossRefPubMedGoogle Scholar
  33. 33.
    Freiberger T, Ravčuková B, Grodecká L, Pikulová Z, Stikarovská D, Pešák S, et al. Sequence variants of the TNFRSF13B gene in Czech CVID and IgAD patients in the context of other populations. Hum Immunol. 2012;73:1147–54.CrossRefPubMedGoogle Scholar
  34. 34.
    Cui CY, Schlessinger D. EDA signaling and skin appendage development. Cell Cycle. 2006;5:2477–83.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Marienfeld RB, Palkowitsch L, Ghosh S. Dimerization of the I kappa B kinase-binding domain of NEMO is required for tumor necrosis factor alpha-induced NF-kappa B activity. Mol Cell Biol. 2006;26:9209–19.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Rushe M, Silvian L, Bixler S, Chen LL, Cheung A, Bowes S, et al. Structure of a NEMO/IKK-associating domain reveals architecture of the interaction site. Structure. 2008;16:798–808.CrossRefPubMedGoogle Scholar
  37. 37.
    Gautheron J, Pescatore A, Fusco F, Esposito E, Yamaoka S, Agou F, et al. Identification of a new NEMO/TRAF6 interface affected in incontinentia pigmenti pathology. Hum Mol Genet. 2010;19:3138–49.CrossRefPubMedGoogle Scholar
  38. 38.
    Bogaert DJ, Dullaers M, Lambrecht BN, Vermaelen KY, De Baere E, Haerynck F. Genes associated with common variable immunodeficiency: one diagnosis to rule them all? J Med Genet. 2016;53:575–90.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Glynis Frans
    • 1
  • Jutte van der Werff Ten Bosch
    • 2
  • Leen Moens
    • 1
  • Rik Gijsbers
    • 3
    • 4
  • Majid Changi-Ashtiani
    • 5
  • Hassan Rokni-Zadeh
    • 6
  • Mohammad Shahrooei
    • 7
    • 8
  • Greet Wuyts
    • 1
  • Isabelle Meyts
    • 9
    • 10
  • Xavier Bossuyt
    • 1
    • 11
    Email author
  1. 1.Department of Microbiology and Immunology, Experimental Laboratory ImmunologyKU LeuvenLeuvenBelgium
  2. 2.Department of Pediatric Hematology, Oncology and ImmunologyUniversity Hospital BrusselsBrusselsBelgium
  3. 3.Leuven Viral Vector CoreKU LeuvenLeuvenBelgium
  4. 4.Department of Pharmaceutical and Pharmacological Sciences, Laboratory for Viral Vector Technology & Gene TherapyKU LeuvenLeuvenBelgium
  5. 5.School of MathematicsInstitute for Research in Fundamental Sciences (IPM)TehranIran
  6. 6.Department of Medical Biotechnology and NanotechnologyZanjan University of Medical SciencesZanjanIran
  7. 7.Department of Microbiology and Immunology, Laboratory of Clinical Bacteriology and MycologyKU LeuvenLeuvenBelgium
  8. 8.Specialized Immunology Laboratory of Dr. ShahrooeiAhvazIran
  9. 9.Department of PediatricsUniversity Hospitals LeuvenLeuvenBelgium
  10. 10.Department of Microbiology and Immunology, Childhood ImmunologyKU LeuvenLeuvenBelgium
  11. 11.Department of Laboratory MedicineUniversity Hospitals LeuvenLeuvenBelgium

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