Brett S J, Blau J, Hughes-Jenkins C M, Rhodes J, Liew F Y, Tite J P
Department of Cell Biology, Wellcome Research Laboratories, Beckenham, Kent, England.
J Immunol. 1991 Aug 1;147(3):984-91.
The characterization of human T cell antigenic sites on influenza A nucleoprotein (NP) is important for subunit vaccine development for either antibody boosting during infection or to stimulate T cell-mediated immunity. To identify such sites, 31 synthetic peptides that cover more than 95% of the amino acid sequence from NP of influenza A/NT/60/68 virus were tested for their ability to stimulate PBL from 42 adult donors. The most frequently recognized region was covered by a peptide corresponding to residues 206-229 of NP, with 20/42 (48%) of responders. In many individuals this was also one of the peptides that stimulated the strongest T cell responses. Other regions that were also frequently recognized were 341-362 by 13/42 (30%), 297-318 by 10/42 (23%), and 182-205 by 9/42 (21%) of individuals. These peptides covered highly conserved regions in NP of influenza A viruses, suggesting that they could be useful in boosting cross-reactive immunity against the known type A virus strains. In order to define the class II restriction molecules used to present particular T cell epitopes, 22 persons from the donor panel were HLA-typed. The majority of persons who expressed DR2, and proliferated to NP also responded to the major immunodominant region 206-229. In addition, this peptide was also immunodominant in the one person expressing DRw13. The observation that recognition of this peptide is associated with DR2 was confirmed by using short term T cell lines and APC from a panel of typed donors. Further results with virus-specific T cell lines and clones and transfected L cells expressing DR molecules showed that DR1 could also present this peptide. Therefore the results suggest that recognition of 206-229 is associated with at least three different DR haplotypes and this may explain the high frequency with which this peptide is recognized in the population. The fine specificity of the response to peptide 206-229 was distinct when presented by DR1- or DR2-expressing APC. The DR1 response was dependent on the N terminus, and the DR2 response was directed to the C terminus of the peptide.
鉴定甲型流感病毒核蛋白（NP）上的人类T细胞抗原位点对于亚单位疫苗的研发很重要，该疫苗可用于在感染期间增强抗体或刺激T细胞介导的免疫反应。为了鉴定这些位点，对覆盖甲型流感病毒A/NT/60/68株NP超过95%氨基酸序列的31条合成肽刺激42名成年供体的外周血淋巴细胞（PBL）的能力进行了检测。最常被识别的区域由对应于NP第206 - 229位残基的肽段覆盖，20/42（48%）的应答者有反应。在许多个体中，这也是刺激最强T细胞反应的肽段之一。其他也常被识别的区域分别是341 - 362位残基，13/42（30%）的个体有反应；297 - 318位残基，10/42（23%）的个体有反应；以及182 - 205位残基，9/42（21%）的个体有反应。这些肽段覆盖了甲型流感病毒NP中的高度保守区域，表明它们可用于增强针对已知甲型病毒株的交叉反应性免疫。为了确定用于呈递特定T细胞表位的Ⅱ类限制分子，对供体组中的22人进行了HLA分型。大多数表达DR2且对NP有增殖反应的人也对主要免疫优势区域206 - 229有反应。此外，该肽段在一名表达DRw13的个体中也是免疫优势的。通过使用来自一组分型供体的短期T细胞系和抗原呈递细胞（APC），证实了对该肽段的识别与DR2相关。对病毒特异性T细胞系和克隆以及表达DR分子的转染L细胞的进一步研究结果表明，DR1也可呈递该肽段。因此，结果表明对206 - 229的识别与至少三种不同的DR单倍型相关，这可能解释了该肽段在人群中被识别的高频率。当由表达DR1或DR2的APC呈递时，对肽段20
