Development of CTL induced vaccine using reverse immunogenetics.

1.我们利用转染了不同HLA - I类基因的RMA-S细胞建立了一种结合实验，以测试一组肽与HLA - I类分子的结合。利用结合实验检测了一组携带来自HIV-1、HCV和HTLV-1的HLA-B^*3501、B^*5101和A^*2402锚定残基的肽。我们鉴定出170个、50个和131个8 ~ 11聚肽分别与HLA-B^*3501、B^*5101和A^*2402结合。我们尝试用27个HLA-B^*3501结合肽诱导两个携带HLA-B^*3501的HLA-1感染供者PBL中的特异性CTL。11个肽诱导特异性CTL。其中10个肽被鉴定为HLA-B^*3501分子呈递的HIV-1 CTL表位。其中9个表位通过特异性CTL克隆的产生得到进一步证实。目前正在研究HLA-B^*5101或A^*2402呈递的HIV-1 CTL表位和HLA-B^*3501或A^*2402呈递的HCV CTL表位的鉴定。

1.We established a binding assay using RMA-S cells transfected with various HLA class I genes in order to test the binding of a panel of peptides to HLA class I molecules. A panel of peptides carrying anchor residues of HLA-B^*3501, B^*5101 and A^*2402 which derived from HIV-1, HCV and HTLV-1 was tested using the binding assay. We identified 170,50 and 131 8-mer to 11-mer peptides which bound to HLA-B^*3501, B^*5101 and A^*2402, respectively.2.We attempted to induce specific CTL in PBL from two HLA-1 infected donors carrying HLA-B^*3501 by 27 HLA-B^*3501 binding peptides. Eleven peptides induced to specific CTL.Ten of these peptides was identified as HIV-1 CTL epitopes presented by HLA-B^*3501 molecules. Nine of these epitopes was further confirmed by generation of the specific CTL clones.3.Identification of both HIV-1 CTL epitopes presented by HLA-B^*5101 or A^*2402 and HCV CTL epitopes presented by HLA-B^*3501 or A^*2402 is under investigation.