Identification of promiscuous epitopes from the mycobacterial 65-kilodalton heat shock protein recognized by human CD4+ T cells of the Mycobacterium leprae memory repertoire
Identification of promiscuous epitopes from the mycobacterial 65-kilodalton heat shock protein recognized by human CD4+ T cells of the Mycobacterium leprae memory repertoire
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DOI：
10.1128/iai.67.11.5683-5689.1999
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发表时间：
1999-11-01
影响因子：
3.1
通讯作者：
Oftung, F
中科院分区：
文献类型：
--
作者：
Mustafa, AS;Lundin, KEA;Oftung, F
通过使用合成肽的方法，我们映射表位的分枝杆菌65-kDa的热休克蛋白（HSP 65）属于麻风分枝杆菌的记忆库的人T细胞的认可。一组HSP 65反应性CD 4（+）T细胞系和克隆是从热灭活M.麻风，然后测试针对包含M.麻风和结核分枝杆菌HSP 65序列。结果表明，所建立的抗原特异性T细胞系和克隆对12种结核分枝杆菌HSP 65肽有应答，其中9种肽代表结核分枝杆菌和分枝杆菌之间的交叉反应表位。麻风HSP 65（氨基酸[aa] 61至75、141至155、151至165、331至345、371至385、411至425、431至445、441至455和501至515）和3种肽（aa 343至355、417至429和522至534）表示M。麻风HSP 65特异性表位。主要组织相容性复合物限制性酶切分析表明，12个多肽中有9个多肽的提呈受自身HLA-DRB 1基因表达的2个HLA-DR分子之一的限制，而3个多肽的序列与M. leprae和M.结核病HSP 65通过多种HLA-DR分子呈递给T细胞：肽（aa 61至75）由HLA-DR 1、-DR 2和-DR 7呈递，肽（aa 141至155）由HLA-DR 2、-DR 7和-DR 53呈递，而HLA-DR 2和-DR 4（Dw 4和Dw 14）都能够将肽（aa 501至515）呈递给T细胞。此外，在增殖测定中响应于这些肽的T细胞系显示出对用相同HSP 65肽脉冲的自体单核细胞/巨噬细胞的细胞毒性活性。总之，我们证明了来自分枝杆菌HSP 65抗原的混杂肽表位可以作为细胞毒性CD 4（+）T细胞的靶点，这些细胞属于人类记忆T细胞库，可以对抗分枝杆菌。麻风病人这些结果表明，这些表位可能用于基于肽的亚单位疫苗的设计，以对抗分枝杆菌疾病。
By using a synthetic peptide approach, we mapped epitopes from the mycobacterial 65-kDa heat shock protein (HSP65) recognized by human T cells belonging to the Mycobacterium leprae memory repertoire. A panel of HSP65 reactive CD4(+) T-cell lines and clones were established from healthy donors 8 years after immunization with heat-killed M. leprae and then tested for proliferative reactivity against overlapping peptides comprising both the M. leprae and Mycobacterium tuberculosis HSP65 sequences. The results showed that the antigen-specific T-cell lines and clones established responded to 12 mycobacterial HSP65 peptides, of which 9 peptides represented epitopes crossreactive between the M tuberculosis and M. leprae HSP65 (amino acids [aa] 61 to 75, 141 to 155, 151 to 165, 331 to 345, 371 to 385, 411 to 425, 431 to 445, 441 to 455, and 501 to 515) and 3 peptides (aa 343 to 355, 417 to 429, and 522 to 534) represented M. leprae HSP65-specific epitopes. Major histocompatibility complex restriction analysis showed that presentation of 9 of the 12 peptides to T cells were restricted by one of the 2 HLA-DR molecules expressed from self HLA-DRB1 genes, whereas 3 peptides with sequences completely identical between the M. leprae and M. tuberculosis HSP65 were presented to T cells by multiple HLA-DR molecules: peptide (aa 61 to 75) was presented by HLA-DR1, -DR2, and -DR7, peptide (aa 141 to 155) was presented by HLA-DR2, -DR7, and -DR53, whereas both HLA-DR2 and -DR4 (Dw4 and Dw14) were able to present peptide (aa 501 to 515) to T cells. In addition, the T-cell lines responding to these peptides in proliferation assays showed cytotoxic activity against autologous monocytes/macrophages pulsed with the same HSP65 peptides. In conclusion, we demonstrated that promiscuous peptide epitopes from the mycobacterial HSP65 antigen can serve as targets for cytotoxic CD4(+) T cells which belong to the human memory T-cell repertoire against M. leprae. The results suggest that such epitopes might be used in the peptide-based design of subunit vaccines against mycobacterial diseases.