Lymph node–targeted DNA engages TBK1/IFN-I–driven innate immunity to induce potent T cell responses and durable memory in mice and NHPs

Adjuvants are immunoactivators capable of shaping the magnitude and quality of antigen-specific immune responses induced by subunit immunization. Presently, there is an acute need for effective adjuvants that safely induce durable and balanced humoral and cellular responses. Here, we engineered a class of Amphiphile (AMP)–modified, immunostimulatory DNA adjuvants designed for targeted delivery to lymph nodes and enhanced stimulation of TANK-binding kinase 1 (TBK1)–mediated danger-sensing pathways to generate strong adaptive immunity and long-term memory with potent recall potential. AMP-DNA adjuvants induced robust interferon type-I (IFN-I)–driven inflammatory environments in mouse and nonhuman primate (NHP) lymph nodes, leading to significantly enhanced cytokine secretion by polyfunctional CD8 + and CD4 + T cells in multiple tissues, as well as strongly elevated T helper cell 1 (T H 1)–associated and neutralizing antibody responses, in the absence of systemic toxicity. These results demonstrate that AMP modification enables lymph node–targeted DNA adjuvants to potently activate IFN-I signaling to generate substantial cellular and humoral responses crucial for vaccine efficacy.