Chemical cross‐linking facilitates antigen uptake and presentation and provides improved protection from Mpox with a dual‐antigen subunit vaccine
Long Chen,
Chao Shang,
Zihao Wang,
Mengzhu Zheng,
Cuiling Zhang,
Dapeng Li,
Zhanqun Yang,
Yuchao Dong,
Yuru Xu,
Yunsheng Yuan,
Shiyong Fan,
Wu Zhong,
Jian Lin,
Xiao Li
Affiliations
Long Chen
Department of Pharmacy Peking University Third Hospital Cancer Center Peking University Third Hospital Beijing China
Chao Shang
Changchun Veterinary Research Institute Chinese Academy of Agricultural Sciences Changchun China
Zihao Wang
National Engineering Research Center for the Emergency Drug Beijing Institute of Pharmacology and Toxicology Beijing China
Mengzhu Zheng
Key Laboratory of Tropical Biological Resources of Ministry of Education Song Li's Academician Workstation of Hainan University School of Pharmaceutical Sciences Hainan University Haikou China
Cuiling Zhang
Changchun Veterinary Research Institute Chinese Academy of Agricultural Sciences Changchun China
Dapeng Li
Changchun Veterinary Research Institute Chinese Academy of Agricultural Sciences Changchun China
Zhanqun Yang
Department of Pharmacy Peking University Third Hospital Cancer Center Peking University Third Hospital Beijing China
Yuchao Dong
National Engineering Research Center for the Emergency Drug Beijing Institute of Pharmacology and Toxicology Beijing China
Yuru Xu
National Engineering Research Center for the Emergency Drug Beijing Institute of Pharmacology and Toxicology Beijing China
Yunsheng Yuan
Engineering Research Center of Cell & Therapeutic Antibody Ministry of Education Shanghai Jiao Tong University School of Pharmacy Shanghai China
Shiyong Fan
National Engineering Research Center for the Emergency Drug Beijing Institute of Pharmacology and Toxicology Beijing China
Wu Zhong
National Engineering Research Center for the Emergency Drug Beijing Institute of Pharmacology and Toxicology Beijing China
Jian Lin
Department of Pharmacy Peking University Third Hospital Cancer Center Peking University Third Hospital Beijing China
Xiao Li
Changchun Veterinary Research Institute Chinese Academy of Agricultural Sciences Changchun China
Abstract Antigen uptake, processing, and presentation are crucial for the immune responses of protein‐based vaccines. Herein, we introduced a reversible chemical cross‐linking strategy to engineer protein antigens, which can be tracelessly removed upon antigen‐presenting cell (APC) uptake and cellular reduction. The chemically cross‐linked antigen proteins presented significantly enhanced uptake and epitope presentation by APC. We applied this strategy to monkeypox virus antigens A29L and A35R to construct dual‐antigen subunit vaccines. Our results revealed that chemical cross‐linking was robust enough to improve both proteins' APC uptake and lymph node accumulation, with each protein being chemically cross‐linked and administered separately. In vivo validation revealed that the chemical cross‐linking of the two antigen proteins improved immune responses, with increases in antigen‐specific antibody and live virus‐neutralizing antibody production. Monkeypox virus challenge experiments revealed that dual‐antigen vaccines prepared via the chemical cross‐linking strategy mitigated tissue damage, reduced the virus load, and extended mouse survival, which proved that the chemical cross‐linking strategy is valuable for protein‐based subunit vaccine development. In consideration of the current threats from the monkeypox virus and potential future emerging pathogens, the chemical cross‐linking strategy provide powerful tools.
