ev70 prototype strain j670  (ATCC)

Journal: Journal of Virology

Article Title: Enterovirus 70 Receptor Utilization Is Controlled by Capsid Residues That Also Regulate Host Range and Cytopathogenicity

doi: 10.1128/jvi.01569-06

Figure Lengend Snippet: FIG. 1. Host range of EV70-Rmk14 and EV70-Dne in cultured cells. One-step growth analysis of viruses was performed in human (HeLa), monkey (LLC-MK2), and murine (L, L-hDAF) cell lines with EV70-Rmk14 (top) or EV70-Dne (bottom) virus at an MOI of 5. Infections were halted at different times postinfection, and virus titers were determined by plaque assay.

Article Snippet: EV70 prototype strain J670/71 was obtained from the American Type Culture Collection (Manassas, VA) and propagated in rhesus monkey kidney LLC-MK2 cells at 37°C.

Techniques: Cell Culture, Virus, Plaque Assay

Journal: Journal of Virology

Article Title: Enterovirus 70 Receptor Utilization Is Controlled by Capsid Residues That Also Regulate Host Range and Cytopathogenicity

doi: 10.1128/jvi.01569-06

Figure Lengend Snippet: FIG. 2. Host range in cell lines derived from human eye and brain. One-step growth analysis of viruses was performed using EV70-Rmk14 (top) or EV70-Dne (bottom) virus to infect (MOI 5) the following eye and brain derived cell lines: HeLa, T98, LLCMK2, U373MG, 15C4, SY5Y, or HCE. Infections were halted at different times postin- fection, and virus titers were determined by plaque assay.

Article Snippet: EV70 prototype strain J670/71 was obtained from the American Type Culture Collection (Manassas, VA) and propagated in rhesus monkey kidney LLC-MK2 cells at 37°C.

Techniques: Derivative Assay, Virus, Plaque Assay

Journal: Journal of Virology

Article Title: Enterovirus 70 Receptor Utilization Is Controlled by Capsid Residues That Also Regulate Host Range and Cytopathogenicity

doi: 10.1128/jvi.01569-06

Figure Lengend Snippet: FIG. 3. Effect on viral replication of enzyme or antibody treatment of cultured cells. HeLa cells were incubated with one of the following as indicated along the x axis: phosphate-buffered saline (PBS [mock treated]), neuraminidase, PI-PLC, or antibodies specific for hapten or the SCR1 or SCR2 domains of the human DAF molecule. Cells were washed and infected with EV70-Rmk14 or EV70-Dne at an MOI of 3. At 24 h postinfection, the total RNA was isolated from infected cells and transferred onto a nitrocellulose membrane for slot blot analysis. Positive-strand viral replication was assessed by hybridization with a radiolabeled negative-strand EV70 RNA probe. The amount of hy- bridized probe was determined with a PhosphorImager and Image- Quant software and reported as the optical density. The data were normalized to the mock (PBS)-treated sample (PBS treatment 100% replication).

Article Snippet: EV70 prototype strain J670/71 was obtained from the American Type Culture Collection (Manassas, VA) and propagated in rhesus monkey kidney LLC-MK2 cells at 37°C.

Techniques: Cell Culture, Incubation, Saline, Infection, Isolation, Membrane, Dot Blot, Hybridization, Software

Journal: Journal of Virology

Article Title: Enterovirus 70 Receptor Utilization Is Controlled by Capsid Residues That Also Regulate Host Range and Cytopathogenicity

doi: 10.1128/jvi.01569-06

Figure Lengend Snippet: FIG. 4. Role of amino acid differences between EV70-Rmk and EV70-Dne viruses in replication in HeLa cells. One-step growth anal- ysis was performed with mutants of EV70-Dne harboring single amino acid substitutions to the EV70-Rmk14 sequence. HeLa cells were infected at an MOI of 5, infections were halted at different times postinfection, and virus titers were determined by plaque assay. (A) Growth analysis of viruses DDDDD (EV70-Dne encoding amino acids K14, M238, L133, P178, and D226), DRRRR (K14K, I238, F133, R178, N226), RDRRR (E14, M238, F133, R178, and N226), RRDRR (E14, I238, L133, R178, and N226), RRRDR (R14, I238, F133, P178, and N226), and RRRRR (EV70-Rmk, E14, I283, F133, R178, and N226). (B) Growth analysis of viruses DDDDD (see above), RDDDD (E14, M238, L133, P178, and D226), DRDDD (K14, I238, L133, P178, and D226), DDRDD (K14, M238, F133, P178, and D226), DDDRD (K14, M238, L133, R178, and D226), and DDDDR (K14, M238, L133, P178, and N226).

Article Snippet: EV70 prototype strain J670/71 was obtained from the American Type Culture Collection (Manassas, VA) and propagated in rhesus monkey kidney LLC-MK2 cells at 37°C.

Techniques: Sequencing, Infection, Virus, Plaque Assay

Journal: Journal of Virology

Article Title: Enterovirus 70 Receptor Utilization Is Controlled by Capsid Residues That Also Regulate Host Range and Cytopathogenicity

doi: 10.1128/jvi.01569-06

Figure Lengend Snippet: FIG. 5. Role of amino acid differences between EV70-Rmk and EV70-Dne viruses in hDAF usage. One-step growth analysis was per- formed using mutants of EV70-Dne virus (described in the legend to Fig. 4). L-hDAF cells were infected at an MOI of 5, infections were halted at different times postinfection, and virus titers were deter- mined by plaque assay.

Article Snippet: EV70 prototype strain J670/71 was obtained from the American Type Culture Collection (Manassas, VA) and propagated in rhesus monkey kidney LLC-MK2 cells at 37°C.

Techniques: Virus, Infection, Plaque Assay

Journal: Journal of Virology

Article Title: Enterovirus 70 Receptor Utilization Is Controlled by Capsid Residues That Also Regulate Host Range and Cytopathogenicity

doi: 10.1128/jvi.01569-06

Figure Lengend Snippet: FIG. 6. Role of amino acid differences between EV70-Rmk and EV70-Dne viruses in cell killing. HeLa cells were infected at an MOI of 3, using EV70-Dne (DDDDD), EV70-Rmk14 (RRRRR), or mu- tant viruses described in the legend to Fig. 4. Infections were halted at different times postinfection, and cells were pelleted by low-speed centrifugation, resuspended in PBS with trypan blue dye, and exam- ined by light microscopy. The percent viability was determined by dividing the number of cells that excluded dye by the number of cells examined.

Article Snippet: EV70 prototype strain J670/71 was obtained from the American Type Culture Collection (Manassas, VA) and propagated in rhesus monkey kidney LLC-MK2 cells at 37°C.

Techniques: Infection, Centrifugation, Light Microscopy

Journal: Journal of Virology

Article Title: Enterovirus 70 Receptor Utilization Is Controlled by Capsid Residues That Also Regulate Host Range and Cytopathogenicity

doi: 10.1128/jvi.01569-06

Figure Lengend Snippet: FIG. 7. Predicted location in the viral capsid of EV70 amino acids that influence host range and cell killing. The known crystallographic structure of BEV-1 was used to predict the locations in the viral capsid of amino acid residues that differ between EV70-Rmk14 and EV70-Dne. Capsid protomer proteins are color coded, with VP1 blue, VP2 yellow, and VP3 red. VP4 is omitted for clarity. Amino acid changes are enumerated in white. (A) Exterior view of a pentamer, comprising five copies each of VP1, VP2, VP3, and VP4 (not shown), revealing the locations of four of the five strain-specific amino acid changes: in the VP1 DE loop, amino acid 133 of EV70 VP1 (F1133L, BEV 1128 in figure), and three in the canyon, including 238 of EV70 VP3 (I3238M, BEV 3240), 178 of EV70 VP1 (R1178P, BEV 1151), and 226 of EV70 VP1 (N1226D, BEV 1219). Residues on only one protomer are labeled. (B) Closer exterior view of canyon and fivefold axis of symmetry. Amino acid changes in all five protomers are labeled.

Article Snippet: EV70 prototype strain J670/71 was obtained from the American Type Culture Collection (Manassas, VA) and propagated in rhesus monkey kidney LLC-MK2 cells at 37°C.

Techniques: Labeling