In Vitro Study of TLR4-NLRP3-Inflammasome Activation in Innate Immune Response

Toll-like receptors (TLRs) are pivotal players in mediating immune responses. TLR4 is the main receptor for LPS, a strong activator of immune cells. LPS/TLR4-dependent pathway, by inducing NF-kappaB activation, is responsible for the release of several mediators, including IL-1beta, one of the most powerful cytokines deeply involved in inflammatory and immune responses. The same pathway is also involved in NLRP3-inflammasome activation, essential for IL-1beta maturation. NLRP3 is a major component of innate immune responses, being a crucial player of host immune defense against virus, bacterial, or fungal infections. NLRP3-inflammasome and IL-1beta hyperactivation have been associated to the pathogenesis of a wide range of disorders and represent therapeutic targets for the development of new treatments of inflammasome-driven inflammatory and autoimmune diseases.Here, we describe an in vitro protocol to induce LPS/TLR4-dependent NLRP3-inflammasome/IL-1beta activation in immune cells, in order to provide a useful assay to study the efficacy of different anti-inflammatory/immune-modulatory agents.

• Mezzasoma, L.
• Schmidt-Weber, C. B.
• Fallarino, F.