Kurane I, Dai L C, Livingston P G, Reed E, Ennis F A
Department of Medicine, University of Massachusetts Medical Center, Worcester 01655.
J Virol. 1993 Oct;67(10):6285-8. doi: 10.1128/JVI.67.10.6285-6288.1993.
We previously reported that the clone JK34 was cross-reactive for dengue virus types 1, 2, 3, and 4 and recognized NS3 (I. Kurane, M. A. Brinton, A. L. Samson, and F. A. Ennis, J. Virol. 65:1823-1828, 1991). In the present experiments, we defined the epitope at the amino acid level, with 93 15-mer overlapping peptides which cover the entire NS3. A peptide 4 which contains amino acids 251 to 265 of NS3 sensitized the autologous B lymphoblastoid cell line (LCL) to the lysis by JK34. The smallest peptide recognized by JK34 was a 10-mer peptide which contains amino acids 255 to 264 (EIVDLMCHAT). A monoclonal antibody to HLA-DP inhibited the lysis of epitope peptide-pulsed autologous LCL by JK34. Genotypic typing revealed that the HLA-DP of this donor is DPA101, DPB10201, which is serologically defined as HLA-DPw2. JK34 lysed peptide 4-pulsed allogeneic LCL which carried HLA-DPw2. These results indicate that HLA-DPw2 is the restriction allele for recognition of this epitope by JK34.
我们之前报道过克隆株JK34对1、2、3和4型登革病毒具有交叉反应性，并能识别NS3（I. 仓根、M. A. 布林顿、A. L. 萨姆森和F. A. 恩尼斯，《病毒学杂志》65:1823 - 1828，1991年）。在本实验中，我们在氨基酸水平上确定了表位，使用了覆盖整个NS3的93个15肽重叠肽。包含NS3第251至265位氨基酸的肽4使自体B淋巴母细胞系（LCL）对JK34介导的裂解敏感。JK34识别的最小肽是一个包含第255至264位氨基酸（EIVDLMCHAT）的10肽。一种针对HLA - DP的单克隆抗体抑制了JK34对表位肽脉冲自体LCL的裂解。基因分型显示该供体的HLA - DP为DPA101、DPB10201，血清学定义为HLA - DPw2。JK34裂解了携带HLA - DPw2的肽4脉冲同种异体LCL。这些结果表明HLA - DPw2是JK34识别该表位的限制等位基因。
