Botten Jason, Alexander Jeff, Pasquetto Valerie, Sidney John, Barrowman Polly, Ting Joey, Peters Bjoern, Southwood Scott, Stewart Barbara, Rodriguez-Carreno Maria P, Mothe Bianca, Whitton J Lindsay, Sette Alessandro, Buchmeier Michael J
Molecular and Integrative Neurosciences Department, The Scripps Research Institute, La Jolla, CA 92037, USA.
J Virol. 2006 Sep;80(17):8351-61. doi: 10.1128/JVI.00896-06.
Recovery from Lassa virus (LASV) infection usually precedes the appearance of neutralizing antibodies, indicating that cellular immunity plays a primary role in viral clearance. To date, the role of LASV-specific CD8(+) T cells has not been evaluated in humans. To facilitate such studies, we utilized a predictive algorithm to identify candidate HLA-A2 supertype epitopes from the LASV nucleoprotein and glycoprotein precursor (GPC) genes. We identified three peptides (GPC(42-50), GLVGLVTFL; GPC(60-68), SLYKGVYEL; and GPC(441-449), YLISIFLHL) that displayed high-affinity binding (< or =98 nM) to HLA-A0201, induced CD8(+) T-cell responses of high functional avidity in HLA-A0201 transgenic mice, and were naturally processed from native LASV GPC in human HLA-A0201-positive target cells. HLA-A0201 mice immunized with either GPC(42-50) or GPC(60-68) were protected against challenge with a recombinant vaccinia virus that expressed LASV GPC. The epitopes identified in this study represent potential diagnostic reagents and candidates for inclusion in epitope-based vaccine constructs. Our approach is applicable to any pathogen with existing sequence data, does not require manipulation of the actual pathogen or access to immune human donors, and should therefore be generally applicable to category A through C agents and other emerging pathogens.
拉沙病毒（LASV）感染的恢复通常先于中和抗体的出现，这表明细胞免疫在病毒清除中起主要作用。迄今为止，LASV特异性CD8(+) T细胞在人类中的作用尚未得到评估。为便于开展此类研究，我们利用一种预测算法从LASV核蛋白和糖蛋白前体（GPC）基因中识别候选HLA - A2超型表位。我们鉴定出三种肽段（GPC(42 - 50)，GLVGLVTFL；GPC(60 - 68)，SLYKGVYEL；以及GPC(441 - 449)，YLISIFLHL），它们与HLA - A0201具有高亲和力结合（≤98 nM），在HLA - A0201转基因小鼠中诱导出高功能亲和力的CD8(+) T细胞反应，并且在人HLA - A0201阳性靶细胞中由天然LASV GPC自然加工而成。用GPC(42 - 50)或GPC(60 - 68)免疫的HLA - A0201小鼠可免受表达LASV GPC的重组痘苗病毒攻击。本研究中鉴定出的表位代表了潜在的诊断试剂以及基于表位的疫苗构建体中的候选物。我们的方法适用于任何具有现有序列数据的病原体，无需对实际病原体进行操作或获取免疫的人类供体，因此应普遍适用于A类至C类病原体及其他新出现的病原体。
