| Reference | ||
|---|---|---|
| Reference Type | Literature | IEDB_Reference:1209 |
| Title | Human CD8+ CTL specific for the mycobacterial major secreted antigen 85A. | |
| Authors | S M Smith; R Brookes; M R Klein; A S Malin; P T Lukey; A S King; G S Ogg; A V Hill; H M Dockrell | |
| Affiliations | Immunology Unit, Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London, United Kingdom. Steven.Smith@bristol.ac.uk. | |
| Journal | J Immunol | |
| Year | 2000 | |
| Abstract | 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. | |
| Curation Last Updated | 2023-11-21 22:16:12 | |
| Epitope | ||
|---|---|---|
| Epitope ID | 21078 | IEDB_epitope:21078 |
| Chemical Type | Linear peptide | |
| Linear Sequence | GLPVEYLQV | |
| Source Molecule Name | Antigen 85-A precursor | |
| Source Organism | Mycobacterium tuberculosis H37Rv (Mycobacterium tuberculosis str. H37Rv) | |
| Starting Position | 48 | |
| Ending Position | 56 | |
| Epitope Reference Details | ||
|---|---|---|
| Epitope Structure Defines | Exact Epitope | |
| Epitope Name | P48-56 | |
| Reference Starting Position | 48 | |
| Reference Ending Position | 56 | |
| Location of Data in Reference | Table 1 | |
| Immunization | ||
|---|---|---|
| Host Organism | Homo sapiens (human) | |
| 1st In Vivo Process | ||
|---|---|---|
| In Vivo Process Type | Administration in vivo | |
| Disease State | healthy | |
| 1st Immunogen | ||
|---|---|---|
| Epitope Relation | Taxonomic Sibling | |
| Object Type | Organism | |
| Organism | Mycobacterium tuberculosis variant bovis BCG | |
| In Vitro Administration | ||
|---|---|---|
| In Vitro Process Type | Restimulation in vitro | |
| Responder Cell Type | T cell | |
| Stimulator Cell Type | PBMC | |
| In Vitro Immunogen | ||
|---|---|---|
| In Vitro Process Type | Taxonomic Sibling | |
| Object Type | Organism | |
| Organism | Mycobacterium tuberculosis variant bovis BCG | |
| Immunization Comments | ||
|---|---|---|
| Immunization Comments | Donors were HLA-A*0201-positive Mycobacterium bovis BCG-vaccinated individuals. PBMC from healthy BCG-vaccinated donors were stimulated for 14 days withlive Mycobacterium bovis BCG in the presence of IL-2 and IL-7 to produceBCG-reactive short-term cell lines. | |
| T Cell Assay | ||
|---|---|---|
| Qualitative Measurement | Positive | |
| Method/Technique | ELISPOT | |
| Measurement of | IFNg release | |
| Measurement Details | ||
|---|---|---|
| Number of Subjects Tested | 5 | |
| Number of Subjects Responded | 5 | |
| Response Frequency (%) | 100 | |
| Effector Cells | ||
|---|---|---|
| Effector Cell Tissue Type | Blood | |
| Effector Cell Type | T cell CD8+ | |
| Effector Cell Culture Conditions | Short Term Restimulated | |
| Antigen Presenting Cells | ||
|---|---|---|
| Cell Tissue Type | Blood | |
| Cell Type | T cell CD8+ | |
| Cell Culture Conditions | Short Term Restimulated | |
| MHC Allele | ||
|---|---|---|
| MHC Allele Name | HLA-A*02:01 | |
| MHC Evidence Code | Not determined | |
| Antigen | ||
|---|---|---|
| Epitope Relation | Epitope | |
| Chemical Type | Linear peptide | |
| Linear Sequence | GLPVEYLQV | |
| Source Molecule Name | Antigen 85-A precursor | |
| Source Organism | Mycobacterium tuberculosis H37Rv (Mycobacterium tuberculosis str. H37Rv) | |
| Starting Position | 48 | |
| Ending Position | 56 | |
| Antigen Details | ||
|---|---|---|
| Antigen Reference Name | P48-56 | |
| Assay Reference Details | ||
|---|---|---|
| Assay Comments by IEDB Curator | No response was found in control subjects. | |
| Location of Assay Data in Reference | Figures 1, 2 and table 1 | |