title: 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.
abstract: BACKGROUND
  The process for obtaining monoclonal antibodies against a specific antigen is very laborious, involves sophisticated technologies and it is not available in most research laboratories. Considering that most cytokines remain partially conserved among species during evolution, the search for antibody cross-reactivity is an important strategy for immunological studies in veterinary medicine. In this context, the amino acid sequence from human and canine cytokines have demonstrated 49-96 % homology, suggesting high probability of cross-reactivity amongst monoclonal antibodies. For this, 17 commercially available anti-human monoclonal antibodies [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 (#1, #2), TNF-α (#1, #2) and TGF-β], were evaluated in vitro for intracellular cytokine detection in a stimulated canine blood culture by flow cytometry and confocal microscopy. Lymphocytes from peripheral blood of healthy and two unhealthy dogs were analyzed.
  
  
  RESULTS
  Eleven anti-human mAbs [IL-1α, IL-4, IL-5, IL-6, IL-8 (#1, #2), IL-12, IL-17A, TNF-α (#1, #2) and TGF-β] cross-reacted against canine intracellular cytokines. The specificity of the assays was not affected after Fc-blocking. Three anti-human cytokine mAbs [IL-4, IL-8 (#2) and TGF-β] when evaluated by confocal microscopy also cross-reacted with intracellular canine cytokines. The identification of human mAbs that cross-reacted with canine cytokines may support their use as immunological biomarkers in veterinary medicine studies.
  
  
  CONCLUSIONS
  The identification of these 11 anti-human cytokine mAbs that cross-reacted with canine cytokines will be useful immunological biomarkers for pathological conditions by flow cytometry and fluorescence microscopy in dogs.
authors.affiliation: Laboratório de Biomarcadores de Diagnóstico e Monitoração, Centro de Pesquisas René Rachou/FIOCRUZ-MG, Belo Horizonte, Minas Gerais, Brazil. marcela.moreira@cpqrr.fiocruz.br.
authors.forename: Marcela L
authors.initials: ML
authors.lastname: Moreira
authors.affiliation: Laboratório de Bacteriologia Aplicada, Escola de Veterinária, Universidade Federal de Minas Gerais, Avenida Antônio Carlos 6627, Pampulha, Belo Horizonte, MG, 31270-901, Brazil. selesdorneles@yahoo.com.br.
authors.forename: Elaine M S
authors.initials: EM
authors.lastname: Dorneles
authors.affiliation: Laboratório de Biomarcadores de Diagnóstico e Monitoração, Centro de Pesquisas René Rachou/FIOCRUZ-MG, Belo Horizonte, Minas Gerais, Brazil. rsoares@cpqrr.fiocruz.br.
authors.forename: Rodrigo P
authors.initials: RP
authors.lastname: Soares
authors.affiliation: Laboratório de Biomarcadores de Diagnóstico e Monitoração, Centro de Pesquisas René Rachou/FIOCRUZ-MG, Belo Horizonte, Minas Gerais, Brazil. camila-pm4@hotmail.com.
authors.forename: Camila P
authors.initials: CP
authors.lastname: Magalhães
authors.affiliation: Laboratório de Biomarcadores de Diagnóstico e Monitoração, Centro de Pesquisas René Rachou/FIOCRUZ-MG, Belo Horizonte, Minas Gerais, Brazil. christianecosta@cpqrr.fiocruz.br.
authors.forename: Christiane
authors.initials: C
authors.lastname: Costa-Pereira
authors.affiliation: Laboratório de Bacteriologia Aplicada, Escola de Veterinária, Universidade Federal de Minas Gerais, Avenida Antônio Carlos 6627, Pampulha, Belo Horizonte, MG, 31270-901, Brazil. alage@vet.ufmg.br.
authors.forename: Andrey P
authors.initials: AP
authors.lastname: Lage
authors.affiliation: Laboratório de Biomarcadores de Diagnóstico e Monitoração, Centro de Pesquisas René Rachou/FIOCRUZ-MG, Belo Horizonte, Minas Gerais, Brazil. andreat@cpqrr.fiocruz.br.
authors.forename: Andréa
authors.initials: A
authors.lastname: Teixeira-Carvalho
authors.affiliation: Laboratório de Biomarcadores de Diagnóstico e Monitoração, Centro de Pesquisas René Rachou/FIOCRUZ-MG, Belo Horizonte, Minas Gerais, Brazil. oamfilho@cpqrr.fiocruz.br.
authors.forename: Olindo A
authors.initials: OA
authors.lastname: Martins-Filho
authors.affiliation: Laboratório de Biomarcadores de Diagnóstico e Monitoração, Centro de Pesquisas René Rachou/FIOCRUZ-MG, Belo Horizonte, Minas Gerais, Brazil. sobreira@cpqrr.fiocruz.br.
authors.forename: Márcio S S
authors.initials: MS
authors.lastname: Araújo
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D003429:Cross Reactions; D016207:Cytokines / Q000276:immunology*; D004285:Dogs; D005434:Flow Cytometry / Q000662:veterinary; D006801:Humans; D008214:Lymphocytes / Q000382:microbiology*; D018613:Microscopy, Confocal / Q000662:veterinary
chemical_list: D000911:Antibodies, Monoclonal; D016207:Cytokines