Protection against lethal HAdV-4 challenge in STAT1 mice by novel human monoclonal antibodies

INTRODUCTION: Human adenovirus serotype 4 (HAdV-4) is an epidemic pathogen associated with severe acute respiratory disease (ARD) in both pediatric and adult populations. Currently, no available vaccine or therapeutic interventions specifically targeting adenoviruses are available. METHODS: In this study, we isolated peripheral blood mononuclear cells (PBMCs) from HAdV-4 infected donors and generated fully human monoclonal antibodies using single-cell PCR technology. The antibodies were first characterized for their neutralization efficacy both in vitro and in vivo. Subsequently, we predicted key functional residues through structural modeling of antigen-antibody complexes and validated their roles via mutagenesis studies. Finally, the mechanism of intracellular neutralization of antibodies was explored. RESULTS: Through systematic screening, we successfully isolated seven antibodies with specific binding activity, among which monoclonal antibodies (mAbs) 2CF4 and 4AC3 exhibited potent neutralizing capacity against HAdV-4. Notably, we modeled adenoviral lethality using Stat1(-/-) transgenic mice, mAb 2CF4 conferred full protection against HAdV-4 infection in Stat1(-/-) transgenic mice. We identified critical amino acid residues, R99, R102 and T104 aa, of mAb 2CF4 by structural prediction of the antigen-antibody complex. Furthermore, the mAb 2CF4 neutralize the HAdV-4 through the interaction with the widely expressed cytoplasmic Fc-binding protein TRIM21. DISCUSSION: Overall, mAb 2CF4 represents a promising candidate for safe and effective prophylactic and therapeutic strategies against HAdV-4 infection.