Article Figures & Data
Figures
- FIG. 1.
Alignment of the amino acid sequences and positions of dominant T-cell epitopes of ESAT-6 (A) and CFP-10 (B) of M. bovis/M. tuberculosis and M. kansasii genotypes I to V (MKA-1 to MKA-V). Amino acids are represented by the one-letter code. Promiscuous recognition of M. bovis-derived ESAT-6 and CFP-10 peptides (defined as giving rise to T-cell responses in ≥50 of the cattle tested that recognized M. bovis ESAT-6 or CFP-10) is represented by open bars above the sequences (27, 31). The proportions of animals (responder frequencies) that recognized these peptides are shown above the respective bars (27, 31). (Adapted from reference 4 with permission of the publisher.)
- FIG. 2.
Recognition of M. bovis and M. kansasii ESAT-6-derived peptides by T cells from cattle with BTB. PBMCs from naturally infected cattle were stimulated with synthetic peptides (see Table 1 for sequence details). The readout system indicates the IFN-γ production measured by the ex vivo ELISPOT assay. The results are expressed as the mean SFCs/well, and the standard errors of the means for all values are <20% of the means. Horizontal lines indicate the cutoff for positivity (10 ΔSFCs/well).
- FIG. 3.
Recognition of M. bovis and M. kansasii CFP-10-derived peptides by T cells from cattle with BTB. PBMCs from naturally infected cattle were stimulated with synthetic peptides (see Table 1 for sequence details). The readout system indicates the IFN-γ production measured by the ex vivo ELISPOT assay. The results are expressed as the mean SFCs/well, and the standard errors of the means for all values are <20% of the means. Horizontal lines indicate the cutoff for positivity (10 ΔSFCs/well).
- FIG. 4.
BoLA class II restriction of the recognition of M. bovis ESAT-6 peptide 3. Cross-reactive recognition of peptides 3, 3A, and 3B in animal 2 (A) was BoLA-DR restricted (B). M. bovis-specific recognition of peptide 3 in animal 1 (C) was BoLA-DQ restricted (D). Anti-BoLA-DR or anti-BoLA-DQ monoclonal antibody was added to peptide 3-stimulated PBMC cultures (B and D). The results are expressed as the mean numbers of IFN-γ-producing SFCs/well ± standard errors of the means. *, P < 0.05 compared to the results for the other groups (Student's t test).
Tables
- TABLE 1.
Peptides used in this study
Protein and peptide Peptide sequencea Species (Mka genotype)b ESAT-6 1 EAYQGVQQKWDATATE M. bovis 1A --------------Q- M. kansasii (I-V) 2 KWDATATELNNALQNL M. bovis 2A ------Q--------- M. kansasii (I-IV) 2B ------Q---S----- M. kansasii (V) 3 LNNALQNLARTISEAG M. bovis 3A --------S------- M. kansasii (I-IV) 3B --S-----S------- M. kansasii (V) CFP-10 4 QEAGNFERISGDLKTQ M. bovis 4A ------------M--- M. kansasii (IV) 5 IDQVESTAGSLQGQWR M. bovis 5A --------A---A--- M. kansasii (I-V) 6 GSLQGQWRGAAGTAAQAA M. bovis 6A A---A------------- M. kansasii (I-V) 6B A---A------A------ M. kansasii (IV) 7 AGTAAQAAVVRFQEAANK M. bovis 7A --A--------------- M. kansasii (IV) 8 VVRFQEAANKQKQELDEI M. bovis 8A ------------A--E-- M. kansasii (I-V) 9 QKQELDEISTNIRQAGVQYS M. bovis 9A --A----E--------------- M. kansasii (I-V) 10 NIRQAGVQYSRADEEQQQ M. bovis 10A ----------K------- M. kansasii (I-V)
