Epidemiological, economic, and programmatic considerations for specific subpopulation risk groups.

<p>Epidemiological, economic, and programmatic considerations for specific subpopulation risk groups.</p

Epidemiological and economics characteristics of various age groups for considering norovirus vaccines¹^,².

<p>Epidemiological and economics characteristics of various age groups for considering norovirus vaccines<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.1001999#t001fn001" target="_blank">¹</a>^,<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.1001999#t001fn002" target="_blank">²</a>.</p

Critical studies to be performed and questions to be answered to advance vaccine development.

<p>Critical studies to be performed and questions to be answered to advance vaccine development.</p

Estimated impact of rotavirus vaccine on hospitalizations and deaths from rotavirus diarrhea among children <5 in Asia

<p><b>Background</b>: Of the 215,000 global deaths from rotavirus estimated in 2013, 41% occur in Asian countries. However, despite a recommendation for global rotavirus vaccination since 2009, only eight countries in Asia have introduced the rotavirus vaccine into their national immunization program as of September 2017. To help policy makers assess the potential value of vaccination, we projected the reduction in rotavirus hospitalizations and deaths following a hypothetical national introduction of rotavirus vaccines in all countries in Asia using data on national-level rotavirus mortality, <5 population, rotavirus hospitalizations rates, routine vaccination coverage, and vaccine effectiveness.</p> <p><b>Methods</b>: To quantify uncertainty, we generated 1,000 simulations of these inputs.</p> <p><b>Results</b>: Our model predicted 710,000 fewer rotavirus hospitalizations, a 49% decrease from the 1,452,000 baseline hospitalizations and 35,000 fewer rotavirus deaths, a 40% decrease from the 88,000 baseline deaths if all 43 Asian countries had introduced rotavirus vaccine. Similar reductions were projected in subanalyses by vaccine introduction status, subregion, and birth cohort size.</p> <p><b>Conclusion</b>: Rotavirus vaccines will substantially reduce morbidity and mortality due to rotavirus infections in Asia.</p

The ability to detect very low FFU of live RV.

<p>Monolayers of MA104 cells were incubated for 24h with 0.2 to 2000 FFU live RV. Uninfected MA104 cells were used as a negative control. Culture supernatant was used to infect new MA104 monolayers, and previously infected monolayers were visualised by FFA. DAPI (blue) indicates cell nuclei and FITC (green) indicates RV infection. Stained monolayers were visualised using Nikon TiE inverted fluorescent microscope. Scale bar = 100μm. Images representative of 3 replicates per MOI for each passage.</p

Inactivation curves of RV and SFV following exposure to γ-irradiation.

<p>SFV and RV samples were exposed to increased doses of γ-irradiation on dry-ice, and the reduction in virus titre was determined by (A) plaque forming assay for SFV, or (B) fluorescent focus assay for RV. All samples tested in triplicate and data presented as mean ± SEM.</p

Neutralising antibody responses induced by γ-RV.

<p>Mice were primed with live RV or γ-RV twice, 2 weeks apart. Serum samples harvested on Day 21 post-2^nd priming, and neutralising ability of immune serum determined by <i>in vitro</i> neutralisation assay. (A) FFU/well determined following incubation of MA104 cells with sera treated-RV at MOI 0.005. RV treated with serial dilutions of HI control or immune sera. PBS-treated RV used to indicate the baseline level of infection. Data presented as mean ± SEM (n = 2), and analysed by One-Way ANOVA (****, p < 0.0001 compared to naïve control sera for each dilution. #, p < 0.05, when directly comparing immune sera groups). (B) Representative fluorescence images of RV infection after treatment with live or γ-RV sera at 1:1280 dilution. DAPI channel (blue) indicates cell nuclei, and FITC channel (green) indicates RV infection. Scale bar = 100 μm.</p

Sterility testing of 50 kGy γ-irradiated RV.

<p>Monolayers of MA104 cells were incubated for 24h with live RV or γ-RV at 4 x 10⁵ FFU-equivalent/well. Culture supernatant was harvested and used to infect new MA104 cell monolayers, and previously infected monolayers stained for RV infection by FFA. DAPI channel (blue) indicates cell nuclei, and FITC channel (green) indicates RV infection. Stained monolayers visualised using Nikon TiE inverted fluorescence microscope. Scale bar = 100 μm. Images representative of 5 replicates per sample for each passage.</p

RV-specific antibody responses.

<p>Mice were primed twice with live RV or γ-RV, 2 weeks apart. Serum samples harvested on Day 14 post-2^nd priming and analysed for RV-specific IgG using ELISA. (A) Serial dilutions of serum samples and absorbance readings at 450/620 nm for total IgG. (B) IgG titres in primed groups calculated relative to cut-off value (dotted line), determined using OD values of serum from control mice. Data presented as mean ± SEM (n = 6), and analysed by unpaired t-test (n.s., not significant) (C) Absorbance at 450/620nm for total IgG, IgG1, and IgG2c in serum at 1:200 dilution by ELISA. Data presented as mean ± SEM (n = 4), analysed by One-Way ANOVA (****, p < 0.0001).</p