Gliadin-Specific CD8+T Cell Responses Restricted by HLA Class I A*0101 and B*0801 Molecules in Celiac Disease Patients.
Initial studies associated the HLA class I A*01 and B*08 alleles with celiac disease (CD) susceptibility. Subsequent analyses showed a primary association with HLA class II alleles encoding for the HLA DQ2.5 molecule. Because of the strong linkage disequilibrium of A*01 and B*08 alleles with the DR3-DQ2.5 haplotype and a recent genome-wide association study indicating that B*08 and B*39 are predisposing genes, the etiologic role of HLA class I in CD pathogenesis needs to be addressed. We screened gliadin proteins (2α-, 2ω-, and 2γ-gliadin) using bioinformatic algorithms for the presence of peptides predicted to bind A*0101 and B*0801 molecules. The top 1% scoring 9- and 10-mer peptides (N= 97, total) were synthesized and tested in binding assays using purified A*0101 and B*0801 molecules. Twenty of ninety-seven peptides bound B*0801 and only 3 of 97 bound A*0101 with high affinity (IC50< 500 nM). These 23 gliadin peptides were next assayed by IFN-γ ELISPOT for recognition in peripheral blood cells of CD patients and healthy controls carrying the A*0101 and/or B*0801 genes and in A*0101/B*0801-CD patients. Ten of the twenty-three peptides assayed recalled IFN-γ responses mediated by CD8+T cells in A*0101/B*0801+patients with CD. Two peptides were restricted by A*0101, and eight were restricted by B*0801. Of note, 50% (5/10) of CD8+T cell epitopes mapped within the γ-gliadins. Our results highlight the value of predicted binding to HLA molecules for identifying gliadin epitopes and demonstrate that HLA class I molecules restrict the anti-gluten T cell response in CD patients.
Institute of Protein Biochemistry-National Research Council, 80131 Naples, Italy.
Stefania
S
Picascia
La Jolla Institute for Allergy and Immunology, La Jolla, CA 92130.
John
J
Sidney
Institute of Food Sciences-National Research Council, 83100 Avellino, Italy.|Department of Translational Medical Science and European Laboratory for Investigation and Food-Induced Diseases, University Federico II, 80131 Naples, Italy; and.
Alessandra
A
Camarca
Institute of Food Sciences-National Research Council, 83100 Avellino, Italy.
Giuseppe
0000-0002-7294-8277
G
Mazzarella
Gastroenterology Unit, Hospital Moscati, 83100 Avellino, Italy.
Nicola
N
Giardullo
Department of Translational Medical Science and European Laboratory for Investigation and Food-Induced Diseases, University Federico II, 80131 Naples, Italy; and.
Luigi
L
Greco
Department of Translational Medical Science and European Laboratory for Investigation and Food-Induced Diseases, University Federico II, 80131 Naples, Italy; and.
Renata
R
Auricchio
Department of Translational Medical Science and European Laboratory for Investigation and Food-Induced Diseases, University Federico II, 80131 Naples, Italy; and.
Salvatore
S
Auricchio
Department of Translational Medical Science and European Laboratory for Investigation and Food-Induced Diseases, University Federico II, 80131 Naples, Italy; and.
Riccardo
R
Troncone
La Jolla Institute for Allergy and Immunology, La Jolla, CA 92130.
Alessandro
0000-0001-7013-2250
A
Sette
Institute of Protein Biochemistry-National Research Council, 80131 Naples, Italy; c.gianfrani@ibp.cnr.it.|Department of Translational Medical Science and European Laboratory for Investigation and Food-Induced Diseases, University Federico II, 80131 Naples, Italy; and.
Carmen
C
Gianfrani
D000293:Adolescent; D000328:Adult; D000465:Algorithms; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002352:Carrier Proteins / Q000276:immunology / Q000502:physiology; D002446:Celiac Disease / Q000235:genetics / Q000276:immunology* / Q000503:physiopathology; D002648:Child; D002675:Child, Preschool; D019295:Computational Biology; D058501:Enzyme-Linked Immunospot Assay; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D005260:Female; D005805:Genes, MHC Class I; D005903:Gliadin / Q000276:immunology*; D005983:Glutens / Q000276:immunology; D015788:HLA-A1 Antigen / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015795:HLA-B8 Antigen / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics / Q000276:immunology; D008297:Male; D008875:Middle Aged; D010455:Peptides / Q000276:immunology* / Q000378:metabolism; D055815:Young Adult
D002352:Carrier Proteins; D018984:Epitopes, T-Lymphocyte; C506470:HLA-A*01:01 antigen; D015788:HLA-A1 Antigen; C509422:HLA-B*08:01 antigen; D015795:HLA-B8 Antigen; D010455:Peptides; D005983:Glutens; D007371:Interferon-gamma; D005903:Gliadin