Human CD8+ CTL specific for the mycobacterial major secreted antigen 85A.
The role of CD8(+) CTL in protection against tuberculosis in human disease is unclear. In this study, we stimulated the peripheral blood mononuclear cells of bacillus Calmette-Guérin (BCG)-vaccinated individuals with live Mycobacterium bovis BCG bacilli to establish short-term cell lines and then pu...
| Main Authors: | , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2000
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| _version_ | 1797085137709563904 |
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| author | Smith, S Brookes, R Klein, MR Malin, A Lukey, P King, A Ogg, G Hill, A Dockrell, H |
| author_facet | Smith, S Brookes, R Klein, MR Malin, A Lukey, P King, A Ogg, G Hill, A Dockrell, H |
| author_sort | Smith, S |
| collection | OXFORD |
| description | The role of CD8(+) CTL in protection against tuberculosis in human disease is unclear. In this study, we stimulated the peripheral blood mononuclear cells of bacillus Calmette-Guérin (BCG)-vaccinated individuals with live Mycobacterium bovis BCG bacilli to establish short-term cell lines and then purified the CD8(+) T cells. A highly sensitive enzyme-linked immunospot (ELISPOT) assay for single cell IFN-gamma release was used to screen CD8(+) T cells with overlapping peptides spanning the mycobacterial major secreted protein, Ag85A. Three peptides consistently induced a high frequency of IFN-gamma responsive CD8(+) T cells, and two HLA-A*0201 binding motifs, P(48-56) and P(242-250), were revealed within the core sequences. CD8(+) T cells responding to the 9-mer epitopes were visualized within fresh blood by ELISPOT using free peptide or by binding of HLA-A*0201 tetrameric complexes. The class I-restricted CD8(+) T cells were potent CTL effector cells that efficiently lysed an HLA-A2-matched monocyte cell line pulsed with peptide as well as autologous macrophages infected with Mycobacterium tuberculosis or recombinant vaccinia virus expressing the whole Ag85A protein. Tetramer assays revealed a 6-fold higher frequency of peptide-specific T cells than IFN-gamma ELISPOT assays, indicating functional heterogeneity within the CD8(+) T cell population. These results demonstrate a previously unrecognized, MHC class I-restricted, CD8(+) CTL response to a major secreted Ag of mycobacteria and supports the use of Ag85A as a candidate vaccine against tuberculosis. |
| first_indexed | 2024-03-07T02:04:52Z |
| format | Journal article |
| id | oxford-uuid:9ea52170-33cf-409d-a627-152b4208db75 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T02:04:52Z |
| publishDate | 2000 |
| record_format | dspace |
| spelling | oxford-uuid:9ea52170-33cf-409d-a627-152b4208db752022-03-27T00:51:28ZHuman CD8+ CTL specific for the mycobacterial major secreted antigen 85A.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:9ea52170-33cf-409d-a627-152b4208db75EnglishSymplectic Elements at Oxford2000Smith, SBrookes, RKlein, MRMalin, ALukey, PKing, AOgg, GHill, ADockrell, HThe role of CD8(+) CTL in protection against tuberculosis in human disease is unclear. In this study, we stimulated the peripheral blood mononuclear cells of bacillus Calmette-Guérin (BCG)-vaccinated individuals with live Mycobacterium bovis BCG bacilli to establish short-term cell lines and then purified the CD8(+) T cells. A highly sensitive enzyme-linked immunospot (ELISPOT) assay for single cell IFN-gamma release was used to screen CD8(+) T cells with overlapping peptides spanning the mycobacterial major secreted protein, Ag85A. Three peptides consistently induced a high frequency of IFN-gamma responsive CD8(+) T cells, and two HLA-A*0201 binding motifs, P(48-56) and P(242-250), were revealed within the core sequences. CD8(+) T cells responding to the 9-mer epitopes were visualized within fresh blood by ELISPOT using free peptide or by binding of HLA-A*0201 tetrameric complexes. The class I-restricted CD8(+) T cells were potent CTL effector cells that efficiently lysed an HLA-A2-matched monocyte cell line pulsed with peptide as well as autologous macrophages infected with Mycobacterium tuberculosis or recombinant vaccinia virus expressing the whole Ag85A protein. Tetramer assays revealed a 6-fold higher frequency of peptide-specific T cells than IFN-gamma ELISPOT assays, indicating functional heterogeneity within the CD8(+) T cell population. These results demonstrate a previously unrecognized, MHC class I-restricted, CD8(+) CTL response to a major secreted Ag of mycobacteria and supports the use of Ag85A as a candidate vaccine against tuberculosis. |
| spellingShingle | Smith, S Brookes, R Klein, MR Malin, A Lukey, P King, A Ogg, G Hill, A Dockrell, H Human CD8+ CTL specific for the mycobacterial major secreted antigen 85A. |
| title | Human CD8+ CTL specific for the mycobacterial major secreted antigen 85A. |
| title_full | Human CD8+ CTL specific for the mycobacterial major secreted antigen 85A. |
| title_fullStr | Human CD8+ CTL specific for the mycobacterial major secreted antigen 85A. |
| title_full_unstemmed | Human CD8+ CTL specific for the mycobacterial major secreted antigen 85A. |
| title_short | Human CD8+ CTL specific for the mycobacterial major secreted antigen 85A. |
| title_sort | human cd8 ctl specific for the mycobacterial major secreted antigen 85a |
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