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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
- Source :
- Journal of Clinical Immunology. 34:331-339
- Publication Year :
- 2014
- Publisher :
- Springer Science and Business Media LLC, 2014.
-
Abstract
- 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). 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. 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. 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.
- Subjects :
- Male
Models, Molecular
Candidate gene
Glycosylation
Genotype
Protein Conformation
T-Lymphocytes
medicine.medical_treatment
DNA Mutational Analysis
Interleukin-10 Receptor alpha Subunit
Molecular Sequence Data
Immunology
Hematopoietic stem cell transplantation
Biology
medicine.disease_cause
Cell Line
Consanguinity
symbols.namesake
medicine
Humans
Immunology and Allergy
Amino Acid Sequence
Age of Onset
Child
Bone Marrow Transplantation
Sanger sequencing
Mutation
Point mutation
Cell Membrane
Hematopoietic Stem Cell Transplantation
Inflammatory Bowel Diseases
Introns
Pedigree
Transplantation
Alternative Splicing
Protein Transport
Phenotype
Treatment Outcome
Real-time polymerase chain reaction
Child, Preschool
symbols
Cancer research
Female
Stem cell
Sequence Alignment
Signal Transduction
Subjects
Details
- ISSN :
- 15732592 and 02719142
- Volume :
- 34
- Database :
- OpenAIRE
- Journal :
- Journal of Clinical Immunology
- Accession number :
- edsair.doi.dedup.....d81c234466292ad5c6b749cbe51fbf37