Mucosal Immunization with a pH-Responsive Nanoparticle Vaccine Induces Protective CD8⁺ Lung-Resident Memory T Cells

Tissue-resident memory T cells (T_RM) patrol nonlymphoid organs and provide superior protection against pathogens that commonly infect mucosal and barrier tissues, such as the lungs, intestine, liver, and skin. Thus, there is a need for vaccine technologies that can induce a robust, protective T_RM response in these tissues. Nanoparticle (NP) vaccines offer important advantages over conventional vaccines; however, there has been minimal investigation into the design of NP-based vaccines for eliciting T_RM responses. Here, we describe a pH-responsive polymeric nanoparticle vaccine for generating antigen-specific CD8⁺ T_RM cells in the lungs. With a single intranasal dose, the NP vaccine elicited airway- and lung-resident CD8⁺ T_RM cells and protected against respiratory virus challenge in both sublethal (vaccinia) and lethal (influenza) infection models for up to 9 weeks after immunization. In elucidating the contribution of material properties to the resulting T_RM response, we found that the pH-responsive activity of the carrier was important, as a structurally analogous non-pH-responsive control carrier elicited significantly fewer lung-resident CD8⁺ T cells. We also demonstrated that dual-delivery of protein antigen and nucleic acid adjuvant on the same NP substantially enhanced the magnitude, functionality, and longevity of the antigen-specific CD8⁺ T_RM response in the lungs. Compared to administration of soluble antigen and adjuvant, the NP also mediated retention of vaccine cargo in pulmonary antigen-presenting cells (APCs), enhanced APC activation, and increased production of T_RM-related cytokines. Overall, these data suggest a promising vaccine platform technology for rapid generation of protective CD8⁺ T_RM cells in the lungs.