Subject 2 had a barely detectable anti-FVIII antibody response de

Subject 2 had a barely detectable anti-FVIII antibody response despite receiving similar FVIII therapy following a traumatic injury. Comprehensive epitope mapping within the FVIII C2 domain using MHC class II (HLA-DRB1*01:01) tetramers indicated Forskolin solubility dmso that T cells from both brothers recognized two overlapping peptides: FVIII 2186-2205 and 2194-2213. T cells recognizing the specific FVIII peptides were subsequently

isolated, cloned and expanded. Verification of antigen specificity of the T-cell clones indicated that their phenotypes differed (Table 7) [32-34]. At 19 weeks after the initial immune response, Subject 1 had two different populations of T cells. By 21 months, these had converted to a TH2 profile suggesting that the TH17 cell lineage played a role only in the early stages of the anti-FVIII immune response. Clones isolated from Subject 2 showed a separate and distinct TH1 lineage and this patient continued to maintain ‘functional immune tolerance’ to FVIII [34]. 19 weeks after initial immune response: TH17/TH1 cells, 3 T-cell clones isolated

TH1/TH2 cells, 2 T-cell clones isolated 21 months after initial immune response: TH2 cells, 8 T-cell clones isolated More recently, our group has conducted T-cell epitope mapping and identification of T-cell phenotypes in patients with severe haemophilia A and inhibitors. MCE公司 A study is currently Selleckchem KPT 330 ongoing with two specific aims: To identify HLA-DRB1-restricted T-cell epitopes in FVIII involved in inhibitor responses of patients with severe haemophilia A (no circulating FVIII). To characterize FVIII-specific T-cell clones and polyclonal lines and investigate phenotypic differences between subjects with persistent inhibitors vs. those who have achieved functional immune tolerance. Some preliminary data are available

from two patients (Subject 3 and Subject 4) with severe haemophilia A and inhibitors but with different responses to ITI therapy. Patient characteristics and some key results of T-cell epitope mapping are summarized in Table 8. Subject 3 was an 18-year-old male who had failed ITI therapy. He had a large F8 gene deletion and was HLA-DRB1 type: *01:01, *10:01. TEGM was conducted to identify T-cell epitopes contributing to the high-titre inhibitor response. MHC class II (HLA-DR) tetramers were loaded with 20-mer peptides spanning the A2, C1 and C2 domains of the FVIII molecule. CD4+ T cells isolated from blood were stimulated with FVIII (either protein or peptides), then expanded in culture with synthetic FVIII peptides. MHC tetramers were used to stain and isolate T-cell clones and polyclonal lines recognizing epitopes restricted to one of his alleles: HLA-DRB1*01:01 or HLA-DRB1*10:01.

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