title: The evaluation of candidate biomarkers of cell-mediated immunity for the diagnosis of Mycobacterium bovis infection in African buffaloes (Syncerus caffer).
abstract: We evaluated commercially available bovine enzyme linked immunosorbent assays (ELISA) and a human IP-10 ELISA to measure IP-10, MIG, MCP-1, MCP-2, MCP-3 and IL1-RA in buffalo plasma in order to identify sensitive markers of the immune response to Mycobacterium bovis-specific peptides. Additionally, we found that all coding mRNA sequences of these cytokines showed very high homology with their homologues in domestic cattle (97-99%) as did the derived amino acid sequences (97-99%). This high sequence homology between cattle and buffaloes supports the use of bovine ELISAs for the detection these cytokines in buffaloes. MCP-1 concentration showed a positive correlation with that of IFN-γ (p=0.0077) and appears to occur in far greater abundance in buffaloes when compared to humans. Using a bovine IP-10 ELISA, levels of this cytokine were found to be significantly increased in antigen-stimulated blood samples from M. bovis test positive buffaloes (p<0.0001) and IP-10 was detected in far greater abundance than IFN-γ. Measurement of IP-10 with this ELISA may prove to be a sensitive marker of M. bovis infection in African buffaloes.
authors.affiliation: DST/NRF Centre of Excellence for Biomedical TB Research/MRC Centre for Tuberculosis Research/Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, PO Box 19063, Tygerberg 7505, South Africa.
authors.forename: Wynand J
authors.initials: WJ
authors.lastname: Goosen
authors.affiliation: Ezemvelo KZN Wildlife, PO Box 25, Mtubatuba 3935, South Africa.
authors.forename: David
authors.initials: D
authors.lastname: Cooper
authors.affiliation: DST/NRF Centre of Excellence for Biomedical TB Research/MRC Centre for Tuberculosis Research/Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, PO Box 19063, Tygerberg 7505, South Africa.
authors.forename: Robin M
authors.initials: RM
authors.lastname: Warren
authors.affiliation: DST/NRF Centre of Excellence for Biomedical TB Research/MRC Centre for Tuberculosis Research/Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, PO Box 19063, Tygerberg 7505, South Africa.
authors.forename: Michele A
authors.initials: MA
authors.lastname: Miller
authors.affiliation: DST/NRF Centre of Excellence for Biomedical TB Research/MRC Centre for Tuberculosis Research/Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, PO Box 19063, Tygerberg 7505, South Africa.
authors.forename: Paul D
authors.initials: PD
authors.lastname: van Helden
authors.affiliation: DST/NRF Centre of Excellence for Biomedical TB Research/MRC Centre for Tuberculosis Research/Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, PO Box 19063, Tygerberg 7505, South Africa. Electronic address: sparsons@sun.ac.za.
authors.forename: Sven D C
authors.initials: SD
authors.lastname: Parsons
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000097:blood*; D002020:Buffaloes / Q000097:blood / Q000276:immunology / Q000382:microbiology*; D002417:Cattle; D054357:Chemokine CXCL10 / Q000097:blood; D054370:Chemokine CXCL9 / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D007111:Immunity, Cellular / Q000276:immunology*; D053590:Interleukin 1 Receptor Antagonist Protein / Q000097:blood; D018945:Monocyte Chemoattractant Proteins / Q000097:blood; D009164:Mycobacterium Infections / Q000097:blood / Q000276:immunology / Q000662:veterinary*; D009163:Mycobacterium bovis / Q000276:immunology*; D018709:Statistics, Nonparametric
chemical_list: D015415:Biomarkers; D054357:Chemokine CXCL10; D054370:Chemokine CXCL9; D053590:Interleukin 1 Receptor Antagonist Protein; D018945:Monocyte Chemoattractant Proteins
keywords: African buffalo; Bovine tuberculosis; Enzyme linked immunosorbent assay; Interferon gamma; Interferon gamma-induced protein 10