Development of therapeutic hepatitis peptide vaccine using a novel T cell receptor cloning system

我们最近报道了hTEC10系统（10天内高效克隆人类TCR），通过该系统我们可以从单个ag特异性人T细胞中获得TCR cdna，并在10天内确定其抗原特异性。在这项研究中，我们应用该系统克隆了慢性肝炎患者的丙型肝炎病毒（HCV）特异性tcr，并比较了克隆的tcr的亲和力。为此，我们首先利用BIMAS系统预测HCV成分（Core、E1、E2、P7、NS2、NS3、NS4、NS5A、NS5B）的HLA-A24限制性候选表位，然后刺激10例HLA-A24与候选肽混合物混合的患者外周血淋巴细胞（pbl）。结果，我们从4例患者的pbl中检测到活化的CD8+ T细胞。我们将克隆CD8+ T细胞的tcr，并比较克隆的tcr的亲和力。

We have recently reported hTEC10 system (human TCR efficient cloning within 10 days) by which we can obtain TCR cDNAs from a single Ag-specific human T cell and determine their antigen specificity within 10 days. In this study, we applied this system to clone human Hepatitis C virus (HCV)-specific TCRs from patients with chronic hepatitis and compare affinities of cloned TCRs.To this end, we first predicted HLA-A24-restricted candidate epitopes of HCV components (Core, E1, E2, P7, NS2, NS3, NS4, NS5A, NS5B) using BIMAS system, then stimulated peripheral blood lymphocytes (PBLs) from the 10 patients who had HLA-A24 allel with the candidate peptide mixtures. As a result, we detected activated CD8+ T cells from PBLs of 4 patients. We are going to clone TCRs of the CD8+ T cells and compare affinities of cloned TCRs.