|CHAUDHARI, ATUL - US Department Of Agriculture (USDA)|
|KIM, WOO - US Department Of Agriculture (USDA)|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 2/20/2018
Publication Date: N/A
Technical Abstract: Interleukin-4 is an important Th2 type cytokine in mammals which regulates growth and differentiation of B Lymphocytes. On the mammalian counterparts, the function of IL-4 orthologue in chickens (chl-IL-4) is less understood. Although the source of chl-IL-4 is not well established, it has been shown to have an integral role in mediating Th2 type immune responses in chickens. Despite its significant role, no monoclonal antibodies (mAbs) against chl-IL-4 have been developed till date. Therefore, in the present study we used recombinant yeast-expressed chl-IL-4 as an immunogen to develop mAbs against chl-IL-4 to establish the basis for understanding the function of chl-IL-4 in regulating Th2 mediated immune responses in chickens. We developed two mAbs against chl-IL-4 and characterized their specificity by western blot, enzyme linked immunosorbent assay (ELISA) and intracellular staining (ICS). These antibodies displayed specific binding with the recombinant chl-IL-4 as well as endogenous chl-IL-4 by western blot, ELISA and ICS. Further, these antibodies neutralized the inhibitory effect of chl-IL-4 on lipopolysaccharide (LPS)-induced nitric oxide (NO) production and expression of inducible nitric oxide synthase (iNOS) in avian machrophage-like HD11 cells, thus confirming that these antibodies can block the chl-IL-4 bioactivity. In conclusion, the mAbs developed in the present study can be useful for understanding the role of chl-IL-4 in the basic concept of Th1 versus Th2 T cell subsets in chickens.