Interleukin 10 (IL10), also known as human cytokine synthesis inhibitory factor (CSIF), is an anti-inflammatory cytokine. IL10 is a cytokine with multiple, pleiotropic, effects in immunoregulation and inflammation. It downregulates the expression of Th1 cytokines, MHC class II antigens, and co-stimulatory molecules on macrophages. It also enhances B cell survival, proliferation, and antibody production. IL10 can block NF-κB activity, and is involved in the regulation of the JAK-STAT signaling pathway. Investigation also has shown that IL-10 predominantly inhibits lipopolysaccharide (LPS) and bacterial product mediated induction of the pro-inflammatory cytokines TNFα, IL-1β, IL-12 and IFNγ secretion from Toll-Like Receptor (TLR) triggered myeloid lineage cells. Besides, Interleukin 10 Receptor Alpha (IL10Ra) has been identified as an interactor of IL10, thus a binding ELISA assay was conducted to detect the interaction of recombinant rat IL10 and recombinant rat IL10Ra. Briefly, IL10 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to IL10Ra-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-IL10 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of IL10 and IL10Ra was shown in Figure 1, and this effect was in a dose dependent manner.
Figure . The binding activity of IL10 with IL10Ra.