zikv strain h pf 2013  (ATCC)

Journal: Pharmaceutics

Article Title: Self-Organized Nanoparticles of Random and Block Copolymers of Sodium 2-(Acrylamido)-2-methyl-1-propanesulfonate and Sodium 11-(Acrylamido)undecanoate as Safe and Effective Zika Virus Inhibitors

doi: 10.3390/pharmaceutics14020309

Figure Lengend Snippet: PAMPS 75 -b-PAaU n block copolymers are potent inhibitors of Zika infection in vitro. ( A ) The inhibition of the ZIKV by PAMPS polymers. ( B ) PAMPS 75 -b-PAaU n block copolymers and P(AMPS 50 -co-AaU 50 ) copolymer at various concentrations in Vero cells. Inhibition of the infection was evaluated by RT-qPCR. Vero cells were infected in the presence of appropriate polymer concentrations for 3 days. Data are shown as logarithmic reduction values (LRV) of ZIKV RNA copy number per milliliter with SEM (error bars). ( C ) CC 50 , IC 50 and selectivity index (SI) values for PAMPS-PAaU block and random copolymers. ( D ) Titration results of supernatants of Vero cells infected in the presence or absence of PAMPS 75 -b-PAaU n block copolymers and P(AMPS 50 -co-AaU 50 ) random copolymer in different concentrations. Data are shown as logarithmic reduction values of supernatant infectivity (TCID 50 ) per milliliter of at least three replications. ( E ) Fluorescence images of cells infected with ZIKV (TCID 50 = 2000/mL) in the presence or absence of the polymers at 25 µg/mL for 48 h. Cell nuclei are shown in blue, actin is shown in red and ZIKV E protein is denoted in green. Scale bar = 50 µm.

Article Snippet: Zika virus (ZIKV, strain H/PF/2013) was acquired from BEI Resources, Manassas, VA, USA.

Techniques: Blocking Assay, Infection, In Vitro, Inhibition, Quantitative RT-PCR, Polymer, Titration, Fluorescence

Journal: Pharmaceutics

Article Title: Self-Organized Nanoparticles of Random and Block Copolymers of Sodium 2-(Acrylamido)-2-methyl-1-propanesulfonate and Sodium 11-(Acrylamido)undecanoate as Safe and Effective Zika Virus Inhibitors

doi: 10.3390/pharmaceutics14020309

Figure Lengend Snippet: PAMPS 75 -b-PAaU 39 inhibits the ZIKV replication cycle by blocking virus attachment. Inhibitory activity of copolymers was tested in Vero cells, as described in the Materials and Methods section: virus inactivation assay ( A ); cell protection assay ( B ); virus entry assay ( C ); virus replication, assembly, and egress assay ( D ). RT-qPCR was used to assess the number of viral RNA copies and to evaluate the inhibition of the infection. The experiments were carried out in triplicate. The data are shown as average values with SEM (error bars). * ( p < 0.05) denotes values that are significantly different from the control. Representative fluorescent images of ZIKV particles on the cell surface ( E ). Vero cells were incubated with ZIKV in the presence or absence of PAMPS 75 -b-PAaU 39 (25 µg/mL) at 4 °C. Cell nuclei are denoted in blue, and ZIKV E protein is denoted in green. The number of ZIKV virions attached to the cells ( F ). The individual counts, including the statistical error of the mean, are shown.

Article Snippet: Zika virus (ZIKV, strain H/PF/2013) was acquired from BEI Resources, Manassas, VA, USA.

Techniques: Blocking Assay, Virus, Activity Assay, Quantitative RT-PCR, Inhibition, Infection, Control, Incubation

Journal: Frontiers in Immunology

Article Title: Functional Profiling of Antibody Immune Repertoires in Convalescent Zika Virus Disease Patients

doi: 10.3389/fimmu.2021.615102

Figure Lengend Snippet: Repertoire-scale functional data for mAb lineages targeting flavivirus antigens. (A) Computational interrogation of yeast display NGS data after 2 rounds of sorting revealed the functional profile of anti-flavivirus antibodies in convalescent donors. Antibodies were screened against three antigens: ZIKV wild-type VLPs [ZIKV (wt)], ZIKV immature VLPs [ZIKV (pr-mut)], and YFV wild-type VLPs [YFV (wt)]. NGS data from Round 2 was used to calculate enrichment ratio and determine antigen specificity while antibody affinity was predicated based on enrichment in Round 3 affinity gates. Antibody isotype was determined from natively paired heavy:light sequence data. (B) Number of isolated mAb lineages and their antigen reactivity profiles for each donor.

Article Snippet: The cDNA sequence encoding the wild-type prM-E protein sequence of ZIKV “H/PF/2013” strain (Genbank #KJ776791) was codon optimized, synthetized, and cloned into plasmid VRC8400 (VRC/NIAID/NIH, Bethesda, MD, USA) at Genscript (Piscataway, NJ, USA) for expression of ZIKV (wt).

Techniques: Functional Assay, Sequencing, Isolation

Journal: Cell host & microbe

Article Title: Zika virus NS3 mimics a cellular 14-3-3-binding motif to antagonize RIG-I- and MDA5-mediated innate immunity

doi: 10.1016/j.chom.2019.09.012

Figure Lengend Snippet: (A) qRT-PCR analysis of the indicated genes in HeLa cells that were transfected for 24 h with FLAG-MDA5 together with GST (negative control), increasing amounts of GST-NS3 from ZIKV (BRA/2015), or MeV V (positive control).

Article Snippet: Plasmids and reagents FLAG-NS3 (ZIKV, strain MR 766) and FLAG-NS3 (ZIKV, strain H/PF/2013) were custom-synthesized by Invitrogen in the pcDNA3.1+ vector.

Techniques: Quantitative RT-PCR, Transfection, Negative Control, Positive Control

Journal: Cell host & microbe

Article Title: Zika virus NS3 mimics a cellular 14-3-3-binding motif to antagonize RIG-I- and MDA5-mediated innate immunity

doi: 10.1016/j.chom.2019.09.012

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Plasmids and reagents FLAG-NS3 (ZIKV, strain MR 766) and FLAG-NS3 (ZIKV, strain H/PF/2013) were custom-synthesized by Invitrogen in the pcDNA3.1+ vector.

Techniques: Purification, Recombinant, Magnetic Beads, Protease Inhibitor, Luciferase, Fractionation, RNA Extraction, Plasmid Preparation, Software, Real-time Polymerase Chain Reaction