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

BackgroundThe 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.ResultsEleven 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.ConclusionThe 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.

Recapitulation of the accessible interface of biopsy-derived canine intestinal organoids to study epithelial-luminal interactions

“…Although dogs are excellent animal models to study human diseases [1], dog studies are often limited by the number of commercially available reagents targeting major proteins shown to be relevant in mice [53,54]. The RNA-ISH technology provides an in situ analysis of biomarkers within the histopathological context of biological samples as they target the mRNA of select proteins [55].…”

Interkingdom Cross-Talk in Times of Stress: Salmonella Typhimurium Grown in the Presence of Catecholamines Inhibits Porcine Immune Functionality in vitro

“…It has been shown earlier that stress has a negative impact on Salmonella recrudescence in pigs by increasing intracellular Salmonella proliferation in macrophages (64). A direct effect on invasiveness and intracellular survival rate of S. Typhimurium by binding of NA to the histidine kinase QseC was demonstrated in another study in mice (65). S. Typhimurium infection in calves was also aggravated by an increase of bacterial proliferation by NA, probably through acting as an iron donor for the bacteria (66).…”

Tolerability of Topical Tocilizumab Eyedrops in Dogs: A Pilot Study

“…Our results should be interpreted in light of the limitations of our study, which include our small sample size, and the fact that the dogs used in this study did not have KCS. In addition, while it has previously been reported that antihuman IL-6 monoclonal antibodies show cross-reactivity with canine IL-6 26 and that there is a high degree of IL-6 gene sequence similarity across species, including dogs, 27 to our knowledge, no studies have directly shown that tocili-zumab cross-reacts with the canine IL-6 receptor. Future larger studies that specifically examine the cross-reactivity of tocilizumab with canine IL-6 receptors and involve longer treatment periods in dogs with DED will be necessary to further examine the effect of topical tocilizumab eyedrop treatment on the ocular surface.…”