A NF-κB-Based High-Throughput Screening for Immune Adjuvants and Inhibitors

The nuclear factor-kappa B (NF-kappa B) family is crucial for regulating immune and inflammatory responses. The activation of the immune cell signaling pathway usually activates NF-kappa B, causing a protective immune response. NF-kappa B can also cause excessive inflammatory responses by activating a cascade reaction of pro-inflammatory mediators such as cytokines. In this study, we used an NF-kappa B luciferase reporter gene system. Out of more than 800 compounds screened, four NF-kappa B agonists were identified with strong activity at nontoxic concentrations. Subsequently, the adjuvant effect was verified on mouse bone marrow-derived dendritic cells (BMDCs) and macrophages RAW264.7. It was found that fostamatinib (R788) disodium increased the production of IL-6, IL-12p40, and TNF-alpha, indicating that R788 disodium could induce the maturation of antigen-presenting cells (APCs). In addition, three compounds were screened to significantly inhibit NF-kappa B at nontoxic doses, including dehydrocostus lactone (DHL)-a known NF-kappa B inhibitor. The results showed that DHL significantly reduced the release of LPS-induced inflammatory cytokines (including TNF-alpha, IL-6, and IL-12). Our findings indicate that the NF-kappa B-based high-throughput screening can be used to discover potential immune adjuvants and anti-inflammatory molecules.

Automated summary

This study utilized a NF-kappa B luciferase reporter gene system to perform high-throughput screening, identifying four NF-kappa B agonists that can activate NF-kappa B at non-toxic concentrations. The adjuvant effect of these compounds was confirmed on mouse immune cells. Additionally, three compounds that significantly inhibit NF-kappa B at non-toxic doses, including a known NF-kappa B inhibitor, were identified.