Generation of Recombinant Authentic Live Attenuated Human Rotavirus Vaccine Strain RIX4414 (Rotarix<sup>®</sup>) from Cloned cDNAs Using Reverse Genetics
Saori Fukuda,
Masanori Kugita,
Kanako Kumamoto,
Yuki Akari,
Yuki Higashimoto,
Shizuko Nagao,
Takayuki Murata,
Tetsushi Yoshikawa,
Koki Taniguchi,
Satoshi Komoto
Affiliations
Saori Fukuda
Department of Virology, Fujita Health University School of Medicine, Toyoake 470-1192, Aichi, Japan
Masanori Kugita
Education and Research Facility of Animal Models for Human Diseases, Fujita Health University, Toyoake 470-1192, Aichi, Japan
Kanako Kumamoto
Education and Research Facility of Animal Models for Human Diseases, Fujita Health University, Toyoake 470-1192, Aichi, Japan
Yuki Akari
Department of Virology, Fujita Health University School of Medicine, Toyoake 470-1192, Aichi, Japan
Yuki Higashimoto
Department of Clinical Microbiology, Fujita Health University School of Medical Sciences, Toyoake 470-1192, Aichi, Japan
Shizuko Nagao
Education and Research Facility of Animal Models for Human Diseases, Fujita Health University, Toyoake 470-1192, Aichi, Japan
Takayuki Murata
Department of Virology, Fujita Health University School of Medicine, Toyoake 470-1192, Aichi, Japan
Tetsushi Yoshikawa
Department of Pediatrics, Fujita Health University School of Medicine, Toyoake 470-1192, Aichi, Japan
Koki Taniguchi
Department of Virology, Fujita Health University School of Medicine, Toyoake 470-1192, Aichi, Japan
Satoshi Komoto
Department of Virology, Fujita Health University School of Medicine, Toyoake 470-1192, Aichi, Japan
The live attenuated human rotavirus vaccine strain RIX4414 (Rotarix®) is used worldwide to prevent severe rotavirus-induced diarrhea in infants. This strain was attenuated through the cell culture passaging of its predecessor, human strain 89-12, which resulted in multiple genomic mutations. However, the specific molecular reasons underlying its attenuation have remained elusive, primarily due to the absence of a suitable reverse genetics system enabling precise genetic manipulations. Therefore, we first completed the sequencing of its genome and then developed a reverse genetics system for the authentic RIX4414 virus. Our experimental results demonstrate that the rescued recombinant RIX4414 virus exhibits biological characteristics similar to those of the parental RIX4414 virus, both in vitro and in vivo. This novel reverse genetics system provides a powerful tool for investigating the molecular basis of RIX4414 attenuation and may facilitate the rational design of safer and more effective human rotavirus vaccines.
