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Results: 3

1.
Figure 3

Figure 3. From: Complex phenotypes in mosquitoes and mice associated with neutralization escape of a Dengue virus type 1 monoclonal antibody.

Phenotype of T329A DENV-1 in AG129 mice. A–C. AG129 mice were infected via an intracranial route with (A) 101, (B) 102, or (C) 103 PFU of wild type or T329A West-Pac-74 DENV-1 and followed for survival. The number of animals for each dose ranged from 5 to 6 per group. No differences were statistically significant. D. AG129 mice were untreated or pre-treated with 500 µg of DENV1-E106 prior to infection with wild type or T329A West-Pac-74 DENV-1. The number of animals for each antibody ranged from 10 to 18 per group. Statistically significant differences are described in the text.

Bimmi Shrestha, et al. Virology. ;427(2):127-134.
2.
Figure 2

Figure 2. From: Complex phenotypes in mosquitoes and mice associated with neutralization escape of a Dengue virus type 1 monoclonal antibody.

Growth analysis and stability of DENV-1 T329A neutralization escape variant. A–B. Multi-step growth curves of wild type or T329A West-Pac-74 DENV-1 in (A) BHK21-15 or (B) C6/36 cells. Cells were infected at an MOI of 0.01 and supernatants were harvested at 24-hour intervals for titration by plaque assay. The data are the average of three independent experiments performed in duplicate and asterisks indicate differences that are statistically significant. C–D. Supernatants were harvested at 96 hours from wild type or T329A DENV-1 infected (C) BHK21-15 or (D) C6/36 cells and used for PRNT analysis to assess stability of the escape mutation in cell culture. The data are representative of two independent experiments performed in duplicate on BHK21-15 cells.

Bimmi Shrestha, et al. Virology. ;427(2):127-134.
3.
Figure 1

Figure 1. Phenotype of the DENV-1 T329A neutralization escape variant. From: Complex phenotypes in mosquitoes and mice associated with neutralization escape of a Dengue virus type 1 monoclonal antibody.

A. Neutralization curves with DENV1-E106 and the parent 16007 DENV-1 strain (genotype 2), the heterologous West-Pac-74 DENV-1 strain (genotype 4), and the T329A West-Pac-74 DENV-1 strain derived directly from passage in cell culture. The data are representative of three independent experiments performed in duplicate on BHK21-15 cells. B–C. Yield of wild type and T329A West-Pac-74 DENV-1 from supernatants of BHK21-15 cells after infection at different input MOI in the (B) absence or (C) presence of DENV1-E106. The data are the average of three independent experiments performed in duplicate and asterisks indicate differences that are statistically significant. Dashed lines indicate the limit of detection of the assay. D–E. Confirmation of resistant phenotype with wild type and mutant DENV-1 RVP. Genetically engineered DENV-1 RVP displaying the structural proteins of (D) West-Pac-74 or (E) 16007 with or without a single amino acid substitution (T329A) were incubated with increasing concentrations of DENV1-E106. F. Neutralization curves of DENV1-E106 with wild type or T329A West-Pac-74 DENV-1 strains derived by site-directed substitution into an infectious cDNA clone. The data are representative of two independent experiments performed in duplicate on BHK21-15 cells. G–I. Genetically engineered DENV-1 RVP (strain West-Pac-74) with single mutations (G) K90R in prM or (H) V83L in E or both mutations (I) K90R + V83L were incubated with increasing concentrations of DENV1-E106 and analyzed by flow cytometry. For D, E, and G–I, representative data of experiments performed on Raji-DCSIGNR cells is shown. Error bars represent the standard error of the mean of duplicate infections.

Bimmi Shrestha, et al. Virology. ;427(2):127-134.

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