Human genome search in celiac disease: mutated gliadin T-cell-like epitope in two human proteins promotes T-cell activation.
Discovery of a number of novel and known human genes whose protein products bear striking similarity to two or more wheat gliadin domains raised the possibility that human intestinal non-HLA peptides homologous to celiac T-cell epitopes could play a role in non-HLA gene specification in celiac disease. Database searching of the entire human genome identified only 11 gut-expressed proteins with high T-cell epitope homology, particularly to the DQ2-gamma-I-gliadin epitope (i.e. TFIIA, FOXJ2 and IgD; mean BestFit quality score=40 versus random value of 24). Others were similar to DQ2-alpha-I-gliadin (i.e. PAX9; BestFit quality 46 versus 20 for random), or DQ2-alpha-II-gliadin (PHLDA1, known in mice as the T-cell death-associated gene; BestFit quality 43 versus 30 for random) epitopes. Among proteins previously screened for gliadin homology, noteworthy was achaete scute homologous protein (DQ2-alpha-I-gliadin; BestFit quality 41 versus 22 for random). With the exception of IgD, all are nuclear factors. Paying particular attention to the position of potential major histocompatibility complex (MHC) anchor residues, several were selected for testing in a DQ2-gamma-I-gliadin-restricted T-cell system. All native 10-mer peptides were inactive, even when deamidated, but V96F substitution of deamidated TFIIA amino acid residues 91-100 stimulated IL-2 release at levels exceeding the wheat gliadin positive control. Also active, but only slightly, was L1009F substitution of AIB3 amino acid residues 1004-1013. PlotSimilarity alignment of TFIIAs from eight species revealed subthreshold similarity score in the peptide region, in contrast to the highly conserved amino and carboxy termini. Molecular modeling of TFIIA[V96F] peptide points to an important juxtaposition of an upwardly projecting phenylalanine residue at peptide position 6 that likely contacts a receptor complementarity-determining region, and a downwardly projecting glutamic acid residue that fits into the shallow MHC P7 pocket. These observations tentatively point to a new multi-gene hypothesis for the initiation of celiac disease in which deamidated free human peptides with T-cell epitope homology (particularly those made more homologous by mutation) escape negative selection, as per deamidation of the HEL(48-62) peptide in the hen egg lysozyme model of autoimmunity. Deamidation following peptide release due to injury triggers inflammation, thereafter repeatedly provoked by dietary gliadin immunodominant peptides concentrated in the proximal small intestine.
Kumar
Rajesh
R
Department of Cell Biology, University of Virginia, Charlottesville, VA 22908-0732, USA.
Eastwood
Amy L
AL
Brown
Milton L
ML
Laurie
Gordon W
GW
D000595:Amino Acid Sequence; D002446:Celiac Disease / Q000235:genetics* / Q000276:immunology; D019295:Computational Biology; D019992:Databases as Topic; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry* / Q000276:immunology*; D020022:Genetic Predisposition to Disease; D015894:Genome, Human; D005903:Gliadin / Q000737:chemistry* / Q000276:immunology; D018698:Glutamic Acid / Q000378:metabolism; D006683:HLA-DQ Antigens / Q000276:immunology; D006801:Humans; D008213:Lymphocyte Activation; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D010649:Phenylalanine / Q000378:metabolism; D011487:Protein Conformation; D017385:Sequence Homology; D013601:T-Lymphocytes / Q000276:immunology*; D035561:Transcription Factor TFIIA; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000276:immunology
D018984:Epitopes, T-Lymphocyte; D006683:HLA-DQ Antigens; C053186:HLA-DQ2 antigen; D010446:Peptide Fragments; D035561:Transcription Factor TFIIA; D014157:Transcription Factors; D018698:Glutamic Acid; D010649:Phenylalanine; D005903:Gliadin