Innate immune defenses are essential for restricting virus replication and for programming the adaptive immune response against infection. Our studies are focused on defining the pathogen recognition receptor interactions and signaling events triggered by viral pathogen associated molecular patterns (PAMPs) to drive innate antiviral immunity and program/enhance the adaptive immune response to RNA virus infection, antiviral therapy, and vaccination. This work has defined the RIG-I-like receptors, including RIG-I, MDA5, and LGP2, as critical factors in the recognition of RNA virus infection and immune protection against infection and disease. We have identified distinct PAMP/RLR interactions that serve to program the outcome of infection and immunity. We are now applying the principles of PAMP/RIG-I interactions to target RIG-I and RLR signaling through small molecule therapeutics aimed at suppressing virus infection through robust induction of innate antiviral immunity, and to serve as vaccine adjuvants to enhance the immune response to vaccination for lasting protection against RNA virus infection. Our preclinical studies demonstrate that targeting RIG-I and RLR signaling offers broad-spectrum antiviral actions. Moreover, small molecule activation of RIG-I serves to greatly enhance the adaptive immune response induced by specific RNA virus vaccines. Thus, RLRs are critical mediators of innate immunity wherein their functions are essential for protection against RNA virus infection. Targeting RLRs through small molecules provides a host-based regimen of antiviral therapy through the actions of PAMP/RIG-I intracellular and tissue response gene expression networks, and offers strong vaccine adjuvant activity through RLR signaling of innate immunity.