Chavarria Miró, GemmaCastellarnau Serra, Montserrat deFuentes, CristinaD'Andrea Rodríguez-Vida, LucíaPérez-Rodríguez, Francisco JavierBeguiristain, NereaBosch, AlbertGuix Arnau, SusanaPintó Solé, Rosa María2021-07-222021-07-222021-02-181664-302Xhttps://hdl.handle.net/2445/179355The available cell-adapted hepatitis A virus (HAV) strains show a very slow replication phenotype hampering the affordable production of antigen. A fast-growing strain characterized by the occurrence of mutations in the internal ribosome entry site (IRES), combined with changes in the codon composition has been selected in our laboratory. A characterization of the IRES activity of this fast-growing strain (HM175-HP; HP) vs. its parental strain (HM175; L0) was assessed in two cell substrates used in vaccine production (MRC-5 and Vero cells) compared with the FRhK-4 cell line in which its selection was performed. The HP-derived IRES was significantly more active than the L0-derived IRES in all cells tested and both IRES were more active in the FRhK-4 cells. The translation efficiency of the HP-derived IRES was also much higher than the L0-derived IRES, particularly, in genes with a HP codon usage background. These results correlated with a higher virus production in a shorter time for the HP strain compared to the L0 strain in any of the three cell lines tested, and of both strains in the FRhK-4 cells compared to Vero and MRC-5 cells. The addition of wortmannin resulted in the increase of infectious viruses and antigen in the supernatant of FRhK-4 infected cells, independently of the strain. Finally, the replication of both strains in a clone of FRhK-4 cells adapted to grow with synthetic sera was optimal and again the HP strain showed higher yields.10 p.application/pdfengcc-by (c) Chavarria-Miró, Gemma et al., 2021https://creativecommons.org/licenses/by/4.0/Virus de l'hepatitis AVacunacióHepatitis A virusVaccinationinfo:eu-repo/semantics/article7116772021-07-22info:eu-repo/semantics/openAccess33679679