HLA-DR-restricted T-cell responses to factor VIII epitopes in a mild haemophilia A family with missense substitution A2201P.
An HLA-DRA-DRB1*0101-restricted T-cell epitope in the factor VIII (FVIII) C2 domain occurred in a mild haemophilia A patient with missense substitution FVIII-A2201P. His T cells responded to synthetic peptides FVIII(2186-2205) and FVIII(2194-2213) (J Thromb Haemost 2007; 5: 2399). T cells from family members with genotype FVIII-A2201P were analysed to determine if FVIII-specific T cells occur in individuals with a haemophilic mutation but no clinically significant inhibitor response. Fluorescent MHC class II tetramers corresponding to subjects'HLA-DRB1 types were loaded with 20-mer peptides and utilized to label antigen-specific CD4+ T cells. T-cell responses to peptides spanning the FVIII-C2 sequence were evaluated. T cells recognizing specific peptides were cloned, and antigen specificity was verified by proliferation assays. Plasma and/or purified IgG samples were tested for FVIII inhibitory activity. CD4+ T cells and T-cell clones from two brothers who shared the DRB1*0101 allele responded to FVIII(2194-2213). A haemophilic cousin's HLA-DRA-DRB1*1104-restricted response to FVIII(2202-2221) was detected only when CD4+CD25+ cells were depleted. A great uncle and two obligate carriers had no detectable FVIII-C2-specific T cells. Concentrated IgG from the brother without a clinical inhibitor response showed a low-titre FVIII inhibitor. FVIII-specific T cells and inhibitory IgG were found in a previously infused, haemophilic subject who had a sub-clinical FVIII inhibitor. CD4+CD25+ depleted T cells from a non-infused haemophilic cousin recognized an overlapping FVIII epitope, indicating a latent HLA-DRA-DRB1*1104-restricted T-cell response to FVIII. Specific T-cell responses to FVIII can occur without clinically significant inhibitors.
Ettinger
R A
RA
Puget Sound Blood Center, BRI Rm 3016, 921 Terry Avenue, Seattle, WA 98104, USA.
James
E A
EA
Kwok
W W
WW
Thompson
A R
AR
Pratt
K P
KP
D019774:Blood Coagulation Factor Inhibitors / Q000276:immunology*; D049109:Cell Proliferation; D018604:Epitope Mapping; D000939:Epitopes; D005167:Factor VII / Q000235:genetics* / Q000276:immunology*; D005838:Genotype; D006684:HLA-DR Antigens / Q000032:analysis / Q000235:genetics*; D059811:HLA-DRB1 Chains; D006467:Hemophilia A / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D020125:Mutation, Missense; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
D019774:Blood Coagulation Factor Inhibitors; D000939:Epitopes; D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains; D005167:Factor VII