Kuwano K, Braciale T J, Ennis F A
Department of Medicine, University of Massachusetts Medical School, Worcester.
Viral Immunol. 1989 Fall;2(3):163-73. doi: 10.1089/vim.1989.2.163.
A cross-reactive cytotoxic T lymphocyte clone was produced by stimulation with a hybrid protein that contained a portion of the NS1 and the HA2 subunit of A/PR/8/34 (H1N1) virus. Transfer of this clone clears virus from the lungs of mice challenged with H1 or H2 viruses. In these experiments we define the location of the protective CTL epitope to the transmembrane portion of the influenza A virus hemagglutinin which is well-conserved on H1 and H2 subtype viruses. The H1 and H2 cross-reactive CTL clone recognized a synthetic peptide of 23 amino acids (anchor peptide) corresponding to the transmembrane domain of the A/PR/8 (H1) HA as well as the comparable anchor peptide of the A/JAP (H2) HA. The anchor peptide of the A/PR/8 HA competed against the anchor peptide of A/JAP HA in cold target inhibition tests. These results indicate that the epitope recognized by the cross-reactive CTL is located on the transmembrane region of both A/PR/8 HA and A/JAP HA. We prepared synthetic peptides to define the epitope within the transmembrane region of A/PR/8 HA which is recognized by a cross reactive CTL clone. The results indicate that residues 518-528 in the transmembrane region of A/PR/8 HA contain the cross-reactive CTL epitope.
用一种包含A/PR/8/34（H1N1）病毒NS1的一部分和HA2亚基的杂交蛋白刺激产生了一个交叉反应性细胞毒性T淋巴细胞克隆。将该克隆转移可清除用H1或H2病毒攻击的小鼠肺中的病毒。在这些实验中，我们将保护性CTL表位的位置确定为甲型流感病毒血凝素的跨膜部分，该部分在H1和H2亚型病毒上高度保守。H1和H2交叉反应性CTL克隆识别对应于A/PR/8（H1）HA跨膜结构域的23个氨基酸的合成肽（锚定肽）以及A/JAP（H2）HA的可比锚定肽。在冷靶抑制试验中，A/PR/8 HA的锚定肽与A/JAP HA的锚定肽相互竞争。这些结果表明，交叉反应性CTL识别的表位位于A/PR/8 HA和A/JAP HA的跨膜区域。我们制备了合成肽以确定A/PR/8 HA跨膜区域内被交叉反应性CTL克隆识别的表位。结果表明，A/PR/8 HA跨膜区域中的518-528位残基包含交叉反应性CTL表位。
