Validation of a commercial canine assay kit to measure pinniped cytokines.
The present study was conducted to assess and validate the cross-reactivity of commercially available multiplex human and canine cytokine kits coupled with the Bio-Plex 200 platform to measure cytokines in three pinniped species, harbor seals (Phoca vitulina), gray seals (Halichoerus grypus), and harp seals (Pagophilus groenlandicus). Cytokines are important small proteins that help direct a proper immune response to pathogens. The human cytokine kit allowed the detection of cytokines in the supernatant of mitogen-stimulated human peripheral blood mononuclear cells, but not in the three pinniped species studied, with the exception of TNFα and GM-CSF. In contrast, the canine cytokine kit appeared to cross-react with the majority of cytokines in the three pinniped species tested, including the pro-inflammatory cytokine TNFα, the Th1 cytokine INFγ, and the Th2 cytokine IL-10, an anti-inflammatory cytokine. In addition, the pro-inflammatory cytokines IL-6 and IL-8 were also measured in all pinniped species. Overall, the Bio-Plex 200 platform and the canine multiplex cytokine kit allowed the successful measurement of potentially clinically important pinniped cytokines. This additional tool may provide veterinarians with additional information to detect sub-clinical signs of inflammation or evidence for immune response, which may not be revealed during regular medical evaluation, e.g. physical examination, hematology, and serum chemistry.
Department of Pathobiology and Veterinary Science, University of Connecticut, 61 North Eagleville Road, Storrs, CT 06269-3089, United States; Center for Environmental Sciences and Engineering, University of Connecticut, 3107 Horsebarn Hill Road, Storrs, CT 06269-4210, United States. Electronic address: milton.levin@uconn.edu.
Milton
M
Levin
Mystic Aquarium, A Division of Sea Research Foundation, 55 Coogan Boulevard, Mystic, CT 06355, United States.
Tracy
T
Romano
University of New England, Marine Animal Rehabilitation Center, 5 Marial Avenue, Biddeford, ME 04005, United States.
Keith
K
Matassa
Department of Pathobiology and Veterinary Science, University of Connecticut, 61 North Eagleville Road, Storrs, CT 06269-3089, United States.
Sylvain
S
De Guise
D000818:Animals; D019411:Clinical Laboratory Techniques / Q000379:methods; D003429:Cross Reactions; D016207:Cytokines / Q000097:blood* / Q000378:metabolism; D004285:Dogs; D005786:Gene Expression Regulation / Q000276:immunology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D046048:Seals, Earless / Q000097:blood*; D012680:Sensitivity and Specificity; D013045:Species Specificity
D016207:Cytokines; D011933:Reagent Kits, Diagnostic
Bio-Plex 200; Canine; Cytokine; Immunology; Pinniped