Endogenous antigen processing promotes mRNA vaccine CD4 ⁺ T cell responses

Lipid nanoparticle (LNP)-encapsulated nucleoside-modified mRNA vaccines elicit robust CD4 ⁺ T cell responses, which are essential for antiviral immunity ^1–3 . While peptides presented to CD4 ⁺ T cells via major histocompatibility complex class II (MHC II) are traditionally thought to be derived from extracellular sources that are processed by antigen presenting cells (APCs) through the classical exogenous pathway ^4,5 , the precise mechanisms of mRNA-LNP vaccine-specific CD4 ⁺ T cell priming remain unknown. Here, we investigated the role of alternative, endogenous antigen presentation pathways ^6,7 in inducing CD4 ⁺ T cell responses to mRNA-LNP vaccines. APCs treated with mRNA-LNP vaccines were consistently superior in activating T cells under conditions of endogenous, rather than exogenous, presentation. Immunization with an mRNA-LNP vaccine that excludes antigen expression in APCs resulted in lower antigen-specific CD4 ⁺ T cell, T follicular helper cell, and antibody responses than mice receiving control vaccine. In contrast, depletion of vaccine antigen from exogenous sources such as muscle cells resulted in little to no reduction in antigen-specific CD4 ⁺ T cells. Our findings demonstrate that direct presentation of endogenous antigen on MHC II is crucial to mRNA-LNP vaccine-induced immune responses and adds to a growing body of literature that redefines the paradigm of MHC II-restricted antigen processing and presentation.