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Results: 4

1.
Figure 3

Figure 3. Growth Characteristics of Recombinant Viruses in Mice. From: Molecular Determinants of Ebola Virus Virulence in Mice.

Groups of 12 mice were inoculated i.p. with 5 FFU (approximately 500 LD50 values for MA-ZEBOV) of representative viruses. On days 1, 2, 3, and 5 postinoculation, selected organs were removed from three infected animals per group. Virus titers in serum (A), spleen (B) and liver (C) were determined in Vero E6 cells by using a focus-forming assay [36].

Hideki Ebihara, et al. PLoS Pathog. 2006 July;2(7):e73.
2.
Figure 1

Figure 1. Molecular Differences among ZEBOV Mouse-Adapted Variants. From: Molecular Determinants of Ebola Virus Virulence in Mice.

(A) Comparison of ZEBOV variants. Pre–MA-ZEBOV differs from wild-type ZEBOV (WT-ZEBOV) by three amino acids in the GP (red triangles) and a silent mutation in the ORF of the VP40 gene (gray triangle). Serial passage of pre–MA-ZEBOV in progressively older mice yielded MA-ZEBOV [14], which contains coding changes in the NP, VP35, VP24, and polymerase (L) (all shown in red triangles). Two nucleotide changes localize to the NCRs at the 3′ end of the VP30 and the 5′ end of the VP24 gene (red triangles); the remaining three modifications in the NP, VP40, and L ORFs are silent (gray triangles).
(B) Nucleotide and amino acid differences among WT-ZEBOV, pre–MA-ZEBOV, MA-ZEBOV, and MA-RG. The nucleotide changes in GP of pre–MA-ZEBOV, compared to WT-ZEBOV, as well as the changes acquired during adaptation of pre–MA-ZEBOV in mice are shown in red.
−, no change.

Hideki Ebihara, et al. PLoS Pathog. 2006 July;2(7):e73.
3.
Figure 4

Figure 4. Effect of Murine Type I IFNs on Recombinant Virus Replication in Mouse Macrophages. From: Molecular Determinants of Ebola Virus Virulence in Mice.

RAW 264.7 cells (mouse peritoneal macrophage-derived cell line) were infected with a multiplicity of infection of 0.05. Cells were untreated (A), treated with murine IFN-α/β (500 units/ml) 2 h postinfection (B), or treated with murine IFN-α/β (500 units/ml) 12 h prior to and again 2 h after virus adsorption (C). Supernatants were collected on days 0, 1, 2, 3, and 4 postinfection and titrated by use of a focus-forming unit assay in Vero E6 cells [36].

Hideki Ebihara, et al. PLoS Pathog. 2006 July;2(7):e73.
4.
Figure 2

Figure 2. Genetic Determinants of Virulence in Adult Mice. From: Molecular Determinants of Ebola Virus Virulence in Mice.

(A) Determination of MLD50 values. Bars indicate the genotype of the viral gene: MA-ZEBOV (red), WT-ZEBOV (blue). The MLD50 values of recombinant viruses were determined by i.p. inoculation of mice (three to six per group) with serial 10-fold dilutions of virus stock and then monitoring of survival rates. Experiments were carried out in duplicate.
(B) Determination of the mean time to death and dose ranges causing morbidity/mortality. The mean time to death of mice inoculated with 10 FFU (approximately 1,000 MLD50 for MA-ZEBOV) are indicated. Differences in the mean time to death for mice infected with various mutants, compared to that of mice infected with MA-ZEBOV, were considered significant when the p-value was <0.05. The dose range for morbidity/mortality was determined by inoculating groups of six to nine mice with the indicated amounts of viruses and monitoring the mice for weight loss and time to death. Survival numbers (dead/total) are color-coded to indicate the severity of infection in infected mice: no disease (black), illness without mortality (green), less than or equal to 50% mortality (purple), greater than 50% mortality (red). *Dead/total. **p < 0.05.

Hideki Ebihara, et al. PLoS Pathog. 2006 July;2(7):e73.

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