HLA-DQA1 and HLA-DQB1 Alleles, Conferring Susceptibility to Celiac Disease and Type 1 Diabetes, Are More Expressed Than Non-Predisposing Alleles and Are Coordinately Regulated
Abstract
1. Introduction
2. Materials and Methods
2.1. Patients Enrollment and Selection of Antigen Presenting Cells
2.2. Monoclonal Antibodies and Flow Cytometry Analysis
2.3. cDNA Cloning and B-LCL Nucleofection
2.4. RNA Quantization
2.5. T Cell Functional Assay
2.6. Statistical Analysis
3. Results
3.1. The DQA1*03 and DQB1*03 mRNA Associated with T1D Risk Were More Abundant than the mRNA of Non-T1D Related Alleles
3.2. Analysis of Co-Regulated Expression of DQA1*05 and DQB1*02 mRNA
3.3. DR7/DR14 B-LCLs Transfected with pDQA105 Efficiently Stimulated Gluten-Specific CD4+ T Cells
4. Discussion
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Code | Cells a | Diagnosis | DQA1 Genotype | DQB1 Genotype | DQ Phenotype | DR Phenotype |
---|---|---|---|---|---|---|
§1 | B-LCL | T1D | *03/*05 | *02/*03 | DQ2.5/DQ8 | DR3/DR4 |
§2 | B-LCL | T1D | *03/*05 | *02/*02 | DQ2.5/DQ2.3 | DR3/DR9 |
§3 | B-LCL | T1D | *05/*05 | *02/*02 | DQ2.5/DQ2.5 | DR3/DR3 |
§4 | B-LCL | T1D with CD | *03/*05 | *02/*03 | DQ2.5/DQ8 | DR3/DR4 |
§5 | B-LCL | T1D with CD | *01/*03 | *05/*03 | DQ8/DQ5 | DR4/DR16 |
§6 | B-LCL | T1D with CD | *01/*05 | *02/*05 | DQ2.5/DQ5 | DR1/DR3 |
§7 | B-LCL | T1D with CD | *02/*05 | *02/*02 | DQ2.5/DQ2.2 | DR3/DR7 |
§8 | B-LCL | T1D with CD | *05/*05 | *02/*02 | DQ2.5/DQ2.5 | DR3/DR3 |
§9 | PBMC | T1D | *01/*05 | *02/*05 | DQ2.5/DQ5 | DR1/DR3 |
§10 | PBMC | T1D | *01/*05 | *02/*06 | DQ2.5/DQ6 | DR3/DR15 |
§11 | PBMC | T1D | *01/*05 | *02/*05 | DQ2.5/DQ5 | DR1/DR3 |
§12 | PBMC | T1D | *01/*03 | *03/*05 | DQ8/DQ5 | DR4/DR1 |
§13 | PBMC | T1D | *03/*05 | *02/*03 | DQ2.5/DQ8 | DR3/DR4 |
§14 | PBMC | T1D | *03/*05 | *02/*03 | DQ2.5/DQ8 | DR3/DR4 |
§15 | PBMC | T1D | *01/*03 | *03/*05 | DQ8/DQ5 | DR4/DR1 |
§16 | PBMC | T1D | *05/*05 | *02/*03 | DQ2.5/DQ7 | DR3/DR5 |
§17 | PBMC | T1D | *01/*05 | *02/*05 | DQ2.5/DQ5 | DR3/DR1 |
§18 | PBMC | T1D | *03/*03 | *03/*03 | DQ8/DQ8 | DR4/DR4 |
§19 | PBMC | T1D | *01/*03 | *03/*05 | DQ8/DQ5 | DR4/DR1 |
§20 | PBMC | T1D | *03/*03 | *03/*03 | DQ8/DQ8 | DR4/DR4 |
§21 | PBMC | T1D | *01/*03 | *03/*05 | DQ8/DQ5 | DR4/DR1 |
Gene | Primers | Sequences 5′ → 3′ |
---|---|---|
β-Actin | ACT-F ACT-R | TCATGAAGTGTGACGTTGACA CCTAGAAGCATTTGCGGTGCAC |
HLA-DQA1*01 | DQA1*01-F DQA1*R | CGGTGGCCTGAGTTCAGCAA GGAGACTTGGAAAACACTGTGACC |
HLA-DQA1*02 | DQA1*02-F DQA1*R | AAGTTGCCTCTGTTCCACAGAC GGAGACTTGGAAAACACTGTGACC |
HLA-DQA1*03 | DQA1*03-F DQA1*R | CTCTGTTCCGCAGATTTAGAAGA GGAGACTTGGAAAACACTGTGACC |
HLA-DQA1*05 | DQA1*05-F DQA1*R | CTCTGTTCCGCAGATTTAGAAGA GGAGACTTGGAAAACACTGTGACC |
HLA-DQB1*02 | DQB1*02-F DQB1*R | TCTTGTGAGCAGAAGCATCT CAGGATCTGGAAGGTCCAGT |
HLA-DQB1*03 | DQB1*03-F DQB1*R | CGGAGTTGGACACGGTGTGC CAGGATCTGGAAGGTCCAGT |
HLA-DQB1*05 | DQB1*05 DQB1*R | ACAACTACGAGGTGGCGTACC CAGGATCTGGAAGGTCCAGT |
HLA-DQB1*06 | DQB1*06-F DQB1*R | CAGATCAAAGTCCGGTGGTTTC CAGGATCTGGAAGGTCCAGT |
Cells | Patients | mRNA Alleles | Fold Change | Cells | Patients | mRNA Alleles | Fold Change |
---|---|---|---|---|---|---|---|
B-LCL | §1 | DQA1*05 = DQA1*03 | 1 | B-LCL | §5 | DQB1*03 > DQB1*05 | 3.2 |
§4 | |||||||
B-LCL | §2 | DQA1*05 > DQA1*03 | 2.7 | B-LCL | §6 | DQB1*02 > DQB1*05 | 4.6 |
B-LCL | §5 | DQA1*03 > DQA1*01 | 3.2 | B-LCL | §1 | DQB1*02 = DQB1*03 | 1 |
§4 | |||||||
B-LCL | §6 | DQA1*05 > DQA1*01 | 3.0 | PBMC | §9 | DQB1*02 > DQB1*05 | 5.6 |
§11 | |||||||
§17 | |||||||
B-LCL | §7 | DQA1*05 > DQA1*02 | 2.7 | PBMC | §10 | DQB1*02 > DQB1*06 | 5.9 |
PBMC | §9 | DQA1*05 > DQA1*01 | 4.8 | PBMC | §19 | DQB1*03 > DQB1*05 | 3.1 |
§10 | §21 | ||||||
§11 | §12 | ||||||
§17 | §15 | ||||||
PBMC | §12 | DQA1*03 > DQA1*01 | 2.9 | PBMC | §13 | DQB1*02 = DQB1*03 | 1 |
§15 | |||||||
§19 | §14 | ||||||
§21 | |||||||
PBMC | §13 | DQA1*05 = DQA1*03 | 1 | PBMC | §16 | DQB1*02 > DQB1*03 | 3.3 |
§14 |
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Farina, F.; Picascia, S.; Pisapia, L.; Barba, P.; Vitale, S.; Franzese, A.; Mozzillo, E.; Gianfrani, C.; Del Pozzo G, G. HLA-DQA1 and HLA-DQB1 Alleles, Conferring Susceptibility to Celiac Disease and Type 1 Diabetes, Are More Expressed Than Non-Predisposing Alleles and Are Coordinately Regulated. Cells 2019, 8, 751. https://doi.org/10.3390/cells8070751
Farina F, Picascia S, Pisapia L, Barba P, Vitale S, Franzese A, Mozzillo E, Gianfrani C, Del Pozzo G G. HLA-DQA1 and HLA-DQB1 Alleles, Conferring Susceptibility to Celiac Disease and Type 1 Diabetes, Are More Expressed Than Non-Predisposing Alleles and Are Coordinately Regulated. Cells. 2019; 8(7):751. https://doi.org/10.3390/cells8070751
Chicago/Turabian StyleFarina, Federica, Stefania Picascia, Laura Pisapia, Pasquale Barba, Serena Vitale, Adriana Franzese, Enza Mozzillo, Carmen Gianfrani, and Giovanna Del Pozzo G. 2019. "HLA-DQA1 and HLA-DQB1 Alleles, Conferring Susceptibility to Celiac Disease and Type 1 Diabetes, Are More Expressed Than Non-Predisposing Alleles and Are Coordinately Regulated" Cells 8, no. 7: 751. https://doi.org/10.3390/cells8070751
APA StyleFarina, F., Picascia, S., Pisapia, L., Barba, P., Vitale, S., Franzese, A., Mozzillo, E., Gianfrani, C., & Del Pozzo G, G. (2019). HLA-DQA1 and HLA-DQB1 Alleles, Conferring Susceptibility to Celiac Disease and Type 1 Diabetes, Are More Expressed Than Non-Predisposing Alleles and Are Coordinately Regulated. Cells, 8(7), 751. https://doi.org/10.3390/cells8070751