![]() An altered cytokine ratio in an individual is considered a marker of disease state such as asthma and atopy. Simultaneous quantification of several substances in a sample makes it possible to calculate the cytokine ratio. In a study by, 15 human cytokines were measured simultaneously in the same sample with a multiplex particle based flow cytometric method, the LabMAP™-assay (Luminex Corporation, Austin, TX, USA). Recently, multiplex cytokine assays have been developed, which measure several cytokines simultaneously in the same sample. Several methods have been developed for cytokine detection, such as enzyme-linked immunosorbent assay (ELISA), enzyme-linked immuno spot assay (ELISPOT), bioassays, polymerase chain reaction (PCR) and intracellular staining. Therefore, the development of new techniques for measuring ruminant cytokines is warranted. A thorough understanding of how the immune system acts during such diseases could lead to the development of new diagnostic methods and/or treatments. Infectious diseases, like mastitis, represent great health problems among dairy cows and sheep. Quantitation of cytokines in biological fluids could be a useful tool in the diagnosis of inflammatory conditions since cytokine patterns give detailed information about the course of an inflammation. TNF-α has an important role as a mediator of the inflammatory response as it induces expression of adhesion molecules on endothelial cells, activates neutrophils and macrophages, and induces production of nitric oxide and complement factors. The inflammatory response during bacterial infections in ruminants is mediated by pro-inflammatory cytokines, such as tumour necrosis factor-a (TNF-a). Resultaten kan tyda på en oförmåga hos antikropparna att korsreagera mellan arter men matrixproteiner kan också ha minskat signalens magnitud. Ingen av de undersökta antikropparna detekterade dock TNF-α i bovina serum- och mjölkprov med kända koncentrationer av TNF-α tidigare bestämda med ELISA-analys. 1111) korsreagerade med ovint rekombinant TNF-α vid koncentrationer över 2,5 ng/ml. Målet med den här studien var därför att med hjälp av partikelbaserad flödescytometri undersöka om 5 kommersiellt tillgängliga antikroppar mot humant TNF-α unde korsreagera med ovint och/eller bovint TNF-a. Eftersom cytokiner har visats vara konserverade genom evolutionen förekommer korsreaktivitet mellan arter. Tyvärr begränsas ofta studier rörande cytokiner som TNF-α hos idisslare av brist på artspecifika reagenser. En viktig cytokin i den tidiga inflammationsreaktionen är tumournecrosis factor-α (TNF-α). Kommersiellt tillgängliga antikroppar mot human Tumour Necrosis Factor-α testade för korsreaktivitet med ovin och bovin Tumour Necrosis Factor-α med hjälp av flödescytometriska analyserĮn grundlig förståelse för immunsystemet inklusive betydelsen av olika cytokiner vid inflammationssjukdomar hos idisslare kan leda till utveckling av nya diagnostiskametoder och behandlingar. The results could be due to inability of the antibodies to cross-react between species, but quenching of the signal by matrix proteins might also have lowered the response. However, none of the antibodies detected TNF-α in bovine milk, or serum containing known concentrations of bovine TNF-α, as earlier determined with ELISA. 1111) showed cross reactivity with ovine recombinant TNF-α in concentrations above 2.5 ng/ml. Therefore, the aim of the present study was to investigate 5 commercially available antibodies against human TNF-α for their ability to cross-react with ovine and/or bovine TNF-α, using a bead-based flow cytometric method. As cytokines have remained rather conserved during evolution, cross-reactivity between animal species may occur. Unfortunately, the number of studies on cytokines, like TNF-α, in ruminants is limited due to a lack of species-specific reagents. Tumour necrosis factor-α (TNF-α) is an important cytokine in the onset of the inflammatory responses. A thorough understanding of the immune system, including the role of different cytokines, during inflammatory diseases in ruminants could lead to the development of new diagnostic methods and treatments. ![]()
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