Proteome-wide screening reveals immunodominance in the CD8 T cell response against classical swine fever virus with antigen-specificity dependent on MHC class I haplotype expression.
Proteome-wide screening reveals immunodominance in the CD8 T cell response against classical swine fever virus with antigen-specificity dependent on MHC class I haplotype expression.
复制标题
DOI：
10.1371/journal.pone.0084246
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Graham SP
中科院分区：
文献类型：
--
作者：
Franzoni G;Kurkure NV;Essler SE;Pedrera M;Everett HE;Bodman-Smith KB;Crooke HR;Graham SP
猪瘟减毒活疫苗免疫后可迅速产生保护作用，这种保护作用与病毒特异性CD8T细胞干扰素-γ反应有关。在这项研究中，我们通过筛选横跨CSFV C株疫苗多蛋白的多肽库来鉴定和表征CD8T细胞表位，从而评估了这种反应的特异性。合成的多肽分别代表12个CSFV蛋白，并用来刺激4头通过C株疫苗免疫CSFV的猪的PBMC，然后用强毒株Brescia株攻击。用流式细胞仪检测CD8T细胞在核心蛋白、E2、Ns2、Ns3和NS5A蛋白的多肽池中均能诱导显著的干扰素γ表达。对这些抗原肽库的解剖表明，在每种情况下，单个离散的抗原肽或一对重叠的抗原肽负责干扰素-γ的诱导。抗原肽或截断衍生物的筛选和滴定确定了以下抗原区：核心241-255 PESRKLEKALLAWA和NS31902-1912 VEYSFIFLDEY，或最小长度抗原肽：E2996-1003YEPRDSYF、NS21223-1230 STVTGIFL和NS5A3070-3078 RVDNALLKF。这些表位在CSFV毒株之间高度保守，并与相关的疫病病毒观察到可变的序列差异。对表位特异的CD8T细胞的鉴定表明，CD107a动员后有细胞毒性的证据，并且相当大比例的细胞除表达干扰素-α外，还表达肿瘤坏死因子-γ。最后，这些免疫显性CD8 T细胞反应的抗原特异性的变异性被证实与不同的MHC I类单倍型的表达有关。此外，C株免疫动物对NS21223-1230 STVTGIFL和NS31902-1912 VEYSFIFLDEY的识别表明，这些多肽可能受到额外单倍型的限制。因此，所确定的抗原区和表位是评估其针对CSFV的疫苗潜力的有吸引力的目标。
Vaccination with live attenuated classical swine fever virus (CSFV) vaccines induces a rapid onset of protection which has been associated with virus-specific CD8 T cell IFN-γ responses. In this study, we assessed the specificity of this response, by screening a peptide library spanning the CSFV C-strain vaccine polyprotein to identify and characterise CD8 T cell epitopes. Synthetic peptides were pooled to represent each of the 12 CSFV proteins and used to stimulate PBMC from four pigs rendered immune to CSFV by C-strain vaccination and subsequently challenged with the virulent Brescia strain. Significant IFN-γ expression by CD8 T cells, assessed by flow cytometry, was induced by peptide pools representing the core, E2, NS2, NS3 and NS5A proteins. Dissection of these antigenic peptide pools indicated that, in each instance, a single discrete antigenic peptide or pair of overlapping peptides was responsible for the IFN-γ induction. Screening and titration of antigenic peptides or truncated derivatives identified the following antigenic regions: core241–255 PESRKKLEKALLAWA and NS31902–1912 VEYSFIFLDEY, or minimal length antigenic peptides: E2996–1003 YEPRDSYF, NS21223–1230 STVTGIFL and NS5A3070–3078 RVDNALLKF. The epitopes are highly conserved across CSFV strains and variable sequence divergence was observed with related pestiviruses. Characterisation of epitope-specific CD8 T cells revealed evidence of cytotoxicity, as determined by CD107a mobilisation, and a significant proportion expressed TNF-α in addition to IFN-γ. Finally, the variability in the antigen-specificity of these immunodominant CD8 T cell responses was confirmed to be associated with expression of distinct MHC class I haplotypes. Moreover, recognition of NS21223–1230 STVTGIFL and NS31902–1912 VEYSFIFLDEY by a larger group of C-strain vaccinated animals showed that these peptides could be restricted by additional haplotypes. Thus the antigenic regions and epitopes identified represent attractive targets for evaluation of their vaccine potential against CSFV.
登录
查看更多内容
影响因子：
3.7
作者：
Graham SP;Everett HE;Haines FJ;Johns HL;Sosan OA;Salguero FJ;Clifford DJ;Steinbach F;Drew TW;Crooke HR
通讯作者：
Crooke HR
影响因子：
3.7
作者：
Costa SM;Yorio AP;Gonçalves AJ;Vidale MM;Costa EC;Mohana-Borges R;Motta MA;Freire MS;Alves AM
通讯作者：
Alves AM
影响因子：
82.9
作者：
Leslie, AJ;Pfafferott, KJ;Goulder, PJR
通讯作者：
Goulder, PJR
影响因子：
5.5
作者：
Graham, Simon P.;Haines, Felicity J.;Crooke, Helen R.
通讯作者：
Crooke, Helen R.
影响因子：
2.2
作者：
Betts, MR;Brenchley, JM;Koup, RA
通讯作者：
Koup, RA