Cross-reactivity of commercially available anti-human monoclonal antibodies with canine cytokines: establishment of a reliable panel to detect the functional profile of peripheral blood lymphocytes by intracytoplasmic staining

• DOI: 10.1186/s13028-015-0142-y
• Publication Date: 2015-09-11T09:09:08Z
• Authors: Marcela L. Moreira; Elaine M. S. Dorn...; Rodrigo P. Soares; Camila P. Magalhães; Christiane Costa-...; Andrey P. Lage; Andréa Teixeira-C...; Olindo A. Martins...; Márcio S. S. Araújo
• Keywords: Proinflammatory cytokine

The process for obtaining monoclonal antibodies against a specific antigen is very laborious, involves sophisticated technologies and it not available in most research laboratories. Considering that cytokines remain partially conserved among species during evolution, the search antibody cross-reactivity an important strategy immunological studies veterinary medicine. In this context, amino acid sequence from human canine have demonstrated 49–96 % homology, suggesting high probability of amongst antibodies. For this, 17 commercially anti-human [IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8 (#1, #2), IL-10, IL-12, IL-13, IL-17A, IFN-γ TNF-α #2) TGF-β], were evaluated vitro intracellular cytokine detection stimulated blood culture by flow cytometry confocal microscopy. Lymphocytes peripheral healthy two unhealthy dogs analyzed. Eleven mAbs TGF-β] cross-reacted cytokines. specificity assays was affected after Fc-blocking. Three [IL-4, (#2) when microscopy also with identification may support their use as biomarkers medicine studies. these 11 will be useful pathological conditions fluorescence dogs.