Journal: Journal of Virology

Article Title: Mutations of two amino acids in VP5 mediate the attenuation of human rotavirus vaccine: evidence from in vitro and in vivo studies

doi: 10.1128/jvi.01067-25

Figure Lengend Snippet: Effect of VP4 of human and simian origin on rotavirus replication in vitro and shedding in vivo . ( a ) Titer of recombinant VP4 mono-reassortments on CDC-9 P11 background in MA104 cells. N = 6. One-way ANOVA was performed comparing mean titers of recombinant strains to rCDC-9 P11. ( b ) Infection of 5-day-old neonatal rats with 1 × 10 7 FFU of the indicated recombinant rotavirus strains. Bodyweight was monitored for 14 days post infection. N = 10 (days 0–5) and N = 5 (days 6–14). ( c ) Shedding in neonatal rats after infection with 1 × 10 7 FFU of the indicated recombinant rotavirus strains. Shedding was measured in rectal swabs by Premier Rotaclone EIA. Two-way ANOVA with multiple test comparison was performed to compare mean shedding after infection with recombinant strains to mean shedding after rCDC-9 P11 infection. ns, not significant ( P ≥ 0.05); * P < 0.05. Shown are mean and SEM for all experiments.

Article Snippet: African green monkey kidney epithelial cell line MA104 (ATCC CRL-2378.1) was passaged in 199 medium (Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS, one shot, Gibco), 100 IU penicillin/mL, 100 μg/mL streptomycin, and 0.292 mg/mL L-glutamine as previously described ( ).

Techniques: In Vitro, In Vivo, Recombinant, Infection, Comparison

Journal: Journal of Virology

Article Title: Mutations of two amino acids in VP5 mediate the attenuation of human rotavirus vaccine: evidence from in vitro and in vivo studies

doi: 10.1128/jvi.01067-25

Figure Lengend Snippet: Growth characteristics of rescued CDC-9 strains with VP4 mutations. ( a ) Schematic overview of rotavirus VP4 gene. Amino acid (AA) mutations from stool/P11 in MA104 cells to P45 in Vero cells are indicated on the top. ( b ) Recombinant CDC-9 stains were generated by transfecting BHK-T7 cells and propagated for up to 4 passages in MA104 cells. All recombinant strains were titered and sequenced to confirm the accuracy of point mutation(s). N = 3. Shown are mean and SEM. Statistical analysis was performed by One-way ANOVA compared to rCDC-9 P11. Ns, not significant P > 0.5. ** P ≤ 0.1. *** P ≤ 0.001.

Article Snippet: African green monkey kidney epithelial cell line MA104 (ATCC CRL-2378.1) was passaged in 199 medium (Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS, one shot, Gibco), 100 IU penicillin/mL, 100 μg/mL streptomycin, and 0.292 mg/mL L-glutamine as previously described ( ).

Techniques: Recombinant, Generated, Mutagenesis