Reference | ||
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Reference Type | Literature | IEDB_Reference:1291 |
Title | Mapping of murine Th1 helper T-Cell epitopes of mycolyl transferases Ag85A, Ag85B, and Ag85C from Mycobacterium tuberculosis. | |
Authors | S D'Souza; V Rosseels; M Romano; A Tanghe; O Denis; F Jurion; N Castiglione; A Vanonckelen; K Palfliet; Kris Huygen | |
Affiliations | Mycobacterial Immunology, Pasteur Institute of Brussels, Brussels, Belgium. | |
Journal | Infect Immun | |
Year | 2003 | |
Abstract | BALB/c (H-2(d)) and C57BL/6 (H-2(b)) mice were infected intravenously with Mycobacterium tuberculosis H37Rv or vaccinated intramuscularly with plasmid DNA encoding each of the three mycolyl transferases Ag85A, Ag85B, and Ag85C from M. tuberculosis. Th1-type spleen cell cytokine secretion of interleukin-2 (IL-2) and gamma interferon (IFN-gamma) was analyzed in response to purified Ag85 components and synthetic overlapping peptides covering the three mature sequences. Tuberculosis-infected C57BL/6 mice reacted strongly to some peptides from Ag85A and Ag85B but not from Ag85C, whereas tuberculosis-infected BALB/c mice reacted only to peptides from Ag85A. In contrast, spleen cells from both mouse strains produced elevated levels of IL-2 and IFN-gamma following vaccination with Ag85A, Ag85B, and Ag85C DNA in response to peptides of the three Ag85 proteins, and the epitope repertoire was broader than in infected mice. Despite pronounced sequence homology, a number of immunodominant regions contained component specific epitopes. Thus, BALB/c mice vaccinated with all three Ag85 genes reacted against the same amino acid region, 101 to 120, that was also immunodominant for Ag85A in M. bovis BCG-vaccinated and tuberculosis-infected H-2(d) haplotype mice, but responses were completely component specific. In C57BL/6 mice, a cross-reactive T-cell response was detected against two carboxy-terminal peptides spanning amino acids 241 to 260 and 261 to 280 of Ag85A and Ag85B. These regions were not recognized at all in C57BL/6 mice vaccinated with Ag85C DNA. Our results underline the need for comparative analysis of all three Ag85 components in future vaccination studies. | |
Curation Last Updated | 2023-08-18 20:03:06 |
Epitope | ||
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Epitope ID | 19579 | IEDB_epitope:19579 |
Chemical Type | Linear peptide | |
Linear Sequence | GFLNPSEGWWPTLIGLAMND | |
Source Molecule Name | Antigen 85-C precursor | |
Source Organism | Mycobacterium tuberculosis H37Rv (Mycobacterium tuberculosis str. H37Rv) | |
Starting Position | 195 | |
Ending Position | 214 |
Epitope Reference Details | ||
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Epitope Structure Defines | Epitope containing region/antigenic site | |
Epitope Name | Ag85C 151-170 | |
Reference Starting Position | 151 | |
Reference Ending Position | 170 | |
Comments | The Swiss-Prot sequence numbering includes the signal sequence. | |
Location of Data in Reference | Table 1 |
Immunization | ||
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Host Organism | Mus musculus C57BL/6 |
1st In Vivo Process | ||
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In Vivo Process Type | Administration in vivo |
Administration Details | ||
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Route | intramuscular | |
Dose Schedule | 3 doses of 100 μg plasmid DNA 3 wks apart |
1st Immunogen | ||
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Epitope Relation | Source Antigen | |
Chemical Type | Protein | |
Molecule Name | Antigen 85-C precursor | |
Organism | Mycobacterium tuberculosis H37Rv (Mycobacterium tuberculosis str. H37Rv) |
Immunization Comments | ||
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Immunization Comments | Immunogen carrier: Plasmid. Mice were immunized with three i.m. injections of 100ug pDNA encoding Ag85 (A, B or C). Spleens were removed 3 weeks after the last DNA immunization. |
T Cell Assay | ||
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Qualitative Measurement | Positive | |
Method/Technique | ELISA | |
Measurement of | IFNg release |
Effector Cells | ||
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Effector Cell Tissue Type | Spleen | |
Effector Cell Type | Splenocyte | |
Effector Cell Culture Conditions | Direct Ex Vivo |
Antigen Presenting Cells | ||
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Cell Tissue Type | Spleen | |
Cell Type | Splenocyte | |
Cell Culture Conditions | Direct Ex Vivo |
Antigen | ||
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Epitope Relation | Epitope | |
Chemical Type | Linear peptide | |
Linear Sequence | GFLNPSEGWWPTLIGLAMND | |
Source Molecule Name | Antigen 85-C precursor | |
Source Organism | Mycobacterium tuberculosis H37Rv (Mycobacterium tuberculosis str. H37Rv) | |
Starting Position | 195 | |
Ending Position | 214 |
Assay Reference Details | ||
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Assay Comments by IEDB Curator | Overlapping 20-mers spanning the length of Ag85 were tested for reactivity using splenocytes from mice immunized with plasmid DNA encoding Ag85. | |
Location of Assay Data in Reference | Figure 2 |