Identification of an HLA-A*0201-restricted T-Cell epitope on the MPT51 protein, a major secreted protein derived from Mycobacterium tuberculosis, by MPT51 overlapping peptide screening
Identification of an HLA-A*0201-restricted T-Cell epitope on the MPT51 protein, a major secreted protein derived from Mycobacterium tuberculosis, by MPT51 overlapping peptide screening
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DOI：
10.1128/iai.01381-07
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发表时间：
2008-04-01
影响因子：
3.1
通讯作者：
Koide, Yukio
中科院分区：
文献类型：
--
作者：
Aoshi, Taiki;Nagata, Toshi;Koide, Yukio
CD 8（+）T细胞在预防结核分枝杆菌感染中起关键作用。我们发现了一个新的HLA-A*0201限制性CD 8（+）T细胞表位上的优势分泌抗原的M。结核病，MPT 51，在HLA-A*0201转基因HHD小鼠中。用基因枪轰击编码MPT 51的质粒DNA免疫HHD小鼠，并分析免疫脾细胞产生的γ干扰素（IFN-γ）。响应于覆盖成熟MPT 51序列的重叠合成肽，仅通过一种肽p51-70刺激脾细胞产生IFN-γ。细胞内IFN-γ和细胞表面CD 4和CD 8染色的三色流式细胞术分析显示，MPT 51 p51-70肽含有免疫显性CD 8（+）T细胞表位。使用计算机算法的进一步分析允许鉴定真正的T细胞表位，p53-62。使用T2细胞的主要组织相容性复合物I类稳定化测定证实该表位结合HLA-A*0201。T细胞能够裂解MPT 51 p53-62肽脉冲的T2细胞。此外，在结核菌素皮肤试验阳性HLA-A*0201（+）健康个体中发现MPT 51 p53-62特异性记忆CD 8（+）T细胞。使用这种HLA-A*0201限制性CD 8（+）T细胞表位分析MPT 51特异性T细胞在M.结核病感染和设计针对结核病的疫苗是可行的。
CD8(+) T cells play a pivotal role in protection against Mycobacterium tuberculosis infection. We identified a novel HLA-A*0201-restricted CD8(+) T-cell epitope on a dominant secreted antigen of M. tuberculosis, MPT51, in HLA-A*0201 transgenic HHD mice. HHD mice were immunized with plasmid DNA encoding MPT51 with gene gun bombardment, and gamma interferon (IFN-gamma) production by the immune splenocytes was analyzed. In response to overlapping synthetic peptides covering the mature MPT51 sequence, the splenocytes were stimulated to produce IFN-gamma by only one peptide, p51-70. Three-color flow cytometric analysis of intracellular IFN-gamma and cell surface CD4 and CD8 staining revealed that the MPT51 p51-70 peptide contains an immunodominant CD8(+) T-cell epitope. Further analysis using computer algorithms permitted identification of a bona fide T-cell epitope, p53-62. A major histocompatibility complex class I stabilization assay using T2 cells confirmed that this epitope binds to HLA-A*0201. The T cells were capable of lysing MPT51 p53-62 peptide-pulsed T2 cells. In addition, MPT51 p53-62-specific memory CD8(+) T cells were found in tuberculin skin test-positive HLA-A*0201(+) healthy individuals. Use of this HLA-A*0201-restricted CD8(+) T-cell epitope for analysis of the role of MPT51-specific T cells in M. tuberculosis infection and for design of vaccines against tuberculosis is feasible.