RBD‐SD1 Nanoparticle Vaccines From DPP4‐Using Merbecoviruses Elicit a Cross‐Reactive Antibody Response but Limited Cross‐Protective Immunity

Coronaviruses within the Merbecovirus subgenus, including Middle East respiratory syndrome coronavirus (MERS-CoV) and its dipeptidyl peptidase 4 (DPP4)-using relatives, pose a persistent zoonotic threat. Efforts to prepare for future Merbecovirus spillover events require vaccines that protect beyond a single virus strain. To evaluate antigenic conservation and cross-protective potential, SpyCatcher-mi3 nanoparticles displaying the receptor-binding domain and subdomain 1 (RBD-SD1) from three DPP4-using merbecoviruses, MERS-CoV, NL140422, and HKU4, were generated. Female mice immunized with these nanoparticle vaccines elicited robust IgG antibody endpoint binding titers and cross-reactive antibody responses against the three merbecoviruses. Only the MERS-CoV RBD-SD1 vaccine, however, elicited neutralizing antibodies against MERS-CoV. While vaccination with MERS-CoV RBD-SD1 reduced lung viral titers in MERS-CoV-challenged human DPP4 mice below the limit of detection, no significant reduction in virus titers was seen in NL140422- and HKU4-RBD-SD1-vaccine-immunized mice. These findings indicate that while the RBD-SD1 interface presents conserved antigenic features sufficient for serological cross-recognition, these epitopes may not be functionally immunodominant for cross-neutralization.