ArticlePoultry science2026
Chicken interleukin-26: characterization and antigen capture enzyme-linked immunosorbent assay.
Article in Poultry science, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Interleukin-26, a cytokine belonging to the IL-10 family, is an inflammatory mediator in mammals and has been shown to modulate T cell proliferation and gene expression in avian species. Given the limited understanding of chIL-26's function at the protein level, our objective was to produce specific mAbs to chIL-26 and create an immunoassay to investigate its biological significance in chickens. We generated five anti-chIL-26 monoclonal antibody clones by using recombinant chIL-26 as an immunogen in mice. Western blot and indirect ELISA analyses confirmed that all five mAbs specifically recognize the recombinant chIL-26 protein. Using a pairing assay, we determined that the combination of capture antibody #10C7 and biotinylated detection antibody #8A5 was the most effective pair for a sandwich ELISA for chIL-26 detection. To further validate a newly developed antigen-capture ELISA chIL-26 detection, we first stimulated chicken macrophage cells with three agonists-Lipopolysaccharides, polyinosinic:polycytidylic acid, and Resquimod-848 at various concentrations. Second, qRT-PCR was used to confirm that LPS induced significant chIL-26 expression. The changes in chIL-26 production over time was monitored using a monoclonal antibody (mAb) combination (#10C7 and biotinylated #8A5). Antigen capture ELISA revealed a significant increase in IL-26 protein secretion, which peaked at 24 hours post-stimulation. This pattern mirrored the expression of chIL-26 mRNA, as detected by qRT-PCR. This assay was also successfully used to detect the changes in IL-26 levels in the serum of chickens infected with Eimeria maxima (E. maxima) and E. tenella. Compared to the uninfected chickens, IL-26 levels in E. maxima-infected chickens began to increase from 1-day post-infection (dpi), peaked at 3 dpi, and then rapidly decreased. In contrast, IL-26 levels in E. tenella-infected chickens peaked at 7 dpi. All newly produced mAbs specific for chIL-26 effectively neutralized the function of IL-26, as measured by cell proliferation assays and IL-1β expression using qRT-PCR analysis. These new anti-chIL-26 mAbs and the antigen-capture ELISA will be valuable tools for further dissecting the role of chIL-26 in poultry inflammatory responses and diseases.
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