Interdisciplinary Graduate Program in Immunology, University of Iowa, Iowa City, Iowa, USA.
J Virol. 2014 Mar;88(6):3135-43. doi: 10.1128/JVI.02139-13. Epub 2013 Dec 26.
Respiratory syncytial virus (RSV) is the most common cause of viral lower respiratory tract infections in infants and children under the age of 5. Studies examining RSV infection in susceptible BALB/c mice indicate that both CD4 and CD8 T cells not only contribute to viral clearance but also facilitate RSV-induced disease. However, efforts to understand the mechanisms by which RSV-specific T cells mediate disease following acute RSV infection have been hampered by the lack of defined RSV-specific T cell epitopes. Using an overlapping peptide library spanning each of the RSV-derived proteins, intracellular cytokine staining for gamma interferon was utilized to identify novel RSV-specific CD4 and CD8 T cell epitopes. Five novel CD8 T cell epitopes were revealed within the RSV fusion (F) protein and glycoprotein (G). In addition, five previously unidentified CD4 T cell epitopes were discovered, including epitopes in the phosphoprotein (P), polymerase protein (L), M2-1 protein, and nucleoprotein (N). Though the initial CD4 T cell epitopes were 15 amino acids in length, synthesis of longer peptides increased the frequency of responding CD4 T cells. Our results indicate that CD4 T cell epitopes that are 17 amino acids in length result in more optimal CD4 T cell stimulation than the commonly used 15-mer peptides.
Respiratory syncytial virus (RSV) is the leading cause of hospitalization for lower respiratory tract infection in children. T cells play a critical role in clearing an acute RSV infection, as well as contributing to RSV-induced disease. Here we examined the breadth of the RSV-specific T cell response, using for the first time an overlapping peptide library spanning the entire viral genome. We identified 5 new CD4 and 5 new CD8 T cell epitopes, including a CD8 T cell epitope within the G protein that was previously believed not to elicit a CD8 T cell response. Importantly, we also demonstrated that the use of longer, 17-mer peptides elicits a higher frequency of responding CD4 T cells than the more commonly used 15-mer peptides. Our results demonstrate the breadth of the CD4 and CD8 T cell response to RSV and demonstrate the importance of using longer peptides when stimulating CD4 T cell responses.
呼吸道合胞病毒（RSV）是 5 岁以下婴儿和儿童下呼吸道感染的最常见原因。研究表明，在易感染的 BALB/c 小鼠中，RSV 感染既涉及 CD4 和 CD8 T 细胞，也涉及 RSV 诱导的疾病。然而，由于缺乏明确的 RSV 特异性 T 细胞表位，理解 RSV 特异性 T 细胞在急性 RSV 感染后介导疾病的机制的努力受到了阻碍。使用跨越 RSV 衍生蛋白的重叠肽文库，通过细胞内细胞因子染色针对γ干扰素，鉴定了新的 RSV 特异性 CD4 和 CD8 T 细胞表位。在 RSV 融合（F）蛋白和糖蛋白（G）中发现了 5 个新的 CD8 T 细胞表位。此外，发现了 5 个以前未鉴定的 CD4 T 细胞表位，包括磷蛋白（P）、聚合酶蛋白（L）、M2-1 蛋白和核蛋白（N）中的表位。尽管最初的 CD4 T 细胞表位长度为 15 个氨基酸，但合成较长的肽增加了应答 CD4 T 细胞的频率。我们的结果表明，长度为 17 个氨基酸的 CD4 T 细胞表位比常用的 15 肽更能刺激 CD4 T 细胞。
呼吸道合胞病毒（RSV）是导致儿童下呼吸道感染住院的主要原因。T 细胞在清除急性 RSV 感染以及导致 RSV 诱导的疾病方面发挥着关键作用。在这里，我们首次使用跨越整个病毒基因组的重叠肽文库来检查 RSV 特异性 T 细胞反应的广度。我们鉴定了 5 个新的 CD4 和 5 个新的 CD8 T 细胞表位，包括先前认为不会引起 CD8 T 细胞反应的 G 蛋白中的 CD8 T 细胞表位。重要的是，我们还证明，使用较长的 17 肽会引起更高频率的应答 CD4 T 细胞，而不是更常用的 15 肽。我们的结果表明了 RSV 对 CD4 和 CD8 T 细胞的反应广度，并证明了当刺激 CD4 T 细胞反应时使用较长的肽的重要性。
