Retinoids inhibit interleukin-12 production in macrophages through physical associations of retinoid X receptor and NF kappa B

Lipopolysaccharide (LPS) increases the production of interleukin-12 (IL-12) from mouse macrophages via a kappa B site within the IL-12 p40 promoter. I n this study, we found that retinoids inhibit this LPS-stimulated productio n of IL-12 in a dose-dependent manner. The NF kappa B components p50 and p6 5 bound retinoid X receptor (RXR) in a ligand-independent manner in vitro, and the interaction interfaces involved the p50 residues 1-245, the p65 res idues 194-441, and the N-terminal A/B/C domains of RXR, Activation of macro phages by LPS resulted in markedly enhanced binding activities to the kappa B site, which significantly decreased upon addition of retinoids, as demon strated by the electrophoretic mobility shift assays. In cotransfections of CV-1 and HeLa cells, RXR also inhibited the NF kappa B transactivation in a ligand-dependent manner, whereas a mutant RXR lacking the AF2 transactiva tion domain, which serves as ligand-dependent binding sites for transcripti on integrators SRC-1 and p300, was without any effect. In addition, coexpre ssion of increasing amounts of SRC-1 or p300 relieved the retinoid-mediated inhibition of the NF kappa B transactivation, From these results, we propo se that retinoid-mediated suppression of the IL-12 production from LPS-acti vated macrophages may involve both inhibition of the NF kappa B-DNA interac tions and competitive recruitment of transcription integrators between NF k appa B and RXR.