Journal: Frontiers in Microbiology

Article Title: A Peptide-Based Virus Inactivator Protects Male Mice Against Zika Virus-Induced Damage of Testicular Tissue

doi: 10.3389/fmicb.2019.02250

Figure Lengend Snippet: Z2 inhibited infection of ZIKV strains with Asian and African lineages in TM4 cells. (A) Sequence and location of Z2 in ZIKV E dimer. Side view of ZIKV E protein in the dimeric conformation is shown [Protein Data Bank: 5IRE ]; DI, DII, DIII, stem and transmembrane domain are shown in red, yellow, blue, orange, and wheat, respectively; the fusion peptide is shown in cyan, and Z2 (421–453) is shown in green. TM4 cells were infected with ZIKV strain SZ01 (B) , MR766 (C) and FLR (D) , treated with Z2 at different concentration, and then viral copies in the supernatant at 48 h were measured by qRT-PCR. Data were presented as means ± SD. (E) Z2-treated ZIKV of different strain lost infectivity on TM4 cells irreversibly. After incubation with Z2 at 37°C for 2 h, ZIKV particles were separated from the unbound Z2 by PEG 8000 to measure their infectivity on TM4 cells. Each sample was tested in triplicate and the experiments were repeated at least once. The data from two independent experiments were presented as mean ± SD.

Article Snippet: ZIKV strains MR766 (#VR1838) and FLR (#VR1844) were obtained from ATCC.

Techniques: Infection, Sequencing, Concentration Assay, Quantitative RT-PCR, Incubation

Journal: Viruses

Article Title: Broadly Active Antiviral Compounds Disturb Zika Virus Progeny Release Rescuing Virus-Induced Toxicity in Brain Organoids

doi: 10.3390/v13010037

Figure Lengend Snippet: Image-based screening on Zika virus (ZIKV)-infected cells identifies TH3289 and TH6744 as active small molecule inhibitors against ZIKV. ( A ) Experimental workflow of phenotypic antiviral testing of small molecule compounds. Nuclei count was used as an indicator of cytotoxicity, viral antibody staining was quantified to assess the percentage of virus infected cells and virus titer was quantified by end-point dilution assay. ( B ) U87 cells were infected with ZIKV (MOI 10) and treated with structural analogs of TH3289 at 10 µM concentration for 48 h. Infected cells were stained for DAPI and ZIKV NS1 and analyzed by high-throughput imaging. Virus titers from supernatants were determined by end-point dilution assay and inhibition of viral titer was calculated relative to dimethyl sulfoxide (DMSO). Data are presented as a mean of two technical replicates per compound performed in n = 1 biological replicate. Representative images from ZIKV-infected U87 cells treated with DMSO or TH3289 (upper image panels; Scale bar equals 100 µm) and corresponding example of titrated ZIKV from the end-point dilution assay (lower image panels) are shown. DAPI in grey, ZIKV NS-1 in magenta. ( C ) The inhibition of viral titers of Hazara virus (HAZV) and ZIKV from both assays was correlated. In the HAZV assay, SW13 cells were infected with HAZV (MOI 10) and treated with 10 μM of compounds from the in-house library for 24 h. Virus titer from supernatant was determined by end-point dilution assay. Cells were stained for DAPI (in blue) and HAZV NP (in green) and analyzed by high-throughput imaging. Data are presented as a mean of two technical replicates per compound performed in n = 1 biological replicate. ( D ) Chemical structures of top hits from ZIKV and HAZV antiviral screenings and their respective inhibition on viral titers.

Article Snippet: For supernatants, the obtained Ct values were converted to the number of ZIKV RNA molecules using a standard curve that was created using Quantitative Synthetic ZIKV RNA from ATCC (ATCC ® VR-3252SDTM).

Techniques: Virus, Infection, Staining, End-point Dilution Assay, Concentration Assay, High Throughput Screening Assay, Imaging, Inhibition

Journal: Viruses

Article Title: Broadly Active Antiviral Compounds Disturb Zika Virus Progeny Release Rescuing Virus-Induced Toxicity in Brain Organoids

doi: 10.3390/v13010037

Figure Lengend Snippet: Series of antiviral compounds have a therapeutic window and rescue ZIKV-induced CPE in cellular models. ( A – E ) U87 cells were infected with ZIKV (MOI 1) and treated with increasing doses of ( A ) TH3289, ( B ) TH5264, ( C ) TH6744, or ( D ) TH5487 for 48 h. Cell viability was determined by nuclei count (in blue), infected cells by ZIKV NS1 staining (in orange) and virus titer by an end-point dilution assay (in black). Data are presented as mean ± SD from n = 3 independent replicates. ( E ) Curve fitting was performed to calculate EC 50 values for living cells, infected cells and viral titer. ( F ) Vero cells were infected with ZIKV (MOI 10) and treated with 12.5 μM TH6744 or DMSO for 48 h. Cells were stained for ZIKV NS1 (in green), ZIKV Capsid (in red) and DAPI (in blue). Images of n = 1 biological replicate. In the overview image, scale bar equals 100 μm and in the close-up 25 μm. ( G ) U87 cells were infected with ZIKV (MOI 1) and treated with 10 μM TH6744 or DMSO for 24, 48, or 72 h. Cells were stained for Pan-Flavi (in green), Golgi marker GM130 (in red), ER marker KDEL (in yellow), and DAPI (in blue). Images of n = 1 biological replicate. Scale bar equals 50 μm. ( H ) Vero cells were infected with ZIKV (MOI 0.1) and treated with increasing doses of TH6744 for 24 h. Viral titer was measured from the supernatants by end-point dilution assay on Vero cells and the development of CPE were monitored by visual assessment. Data presented as mean ± SD from n = 2 biological replicates.

Article Snippet: For supernatants, the obtained Ct values were converted to the number of ZIKV RNA molecules using a standard curve that was created using Quantitative Synthetic ZIKV RNA from ATCC (ATCC ® VR-3252SDTM).

Techniques: Infection, Staining, Virus, End-point Dilution Assay, Marker

Journal: Viruses

Article Title: Broadly Active Antiviral Compounds Disturb Zika Virus Progeny Release Rescuing Virus-Induced Toxicity in Brain Organoids

doi: 10.3390/v13010037

Figure Lengend Snippet: TH6744 and TH5487 treatment reverses ZIKV-induced neurotoxicity and limits ZIKV propagation in three-dimensional (3D) cerebral organoids. ( A ) Experimental workflow of ZIKV infection on brain organoid model. ( B – F ) Organoids were infected with ZIKV (2 × 10 4 particles/organoid) followed by treatment with 5 μM TH6744, 5 μM TH5487 or DMSO. ( B ) Organoids were dissociated to single cells at 7 or 10 dpi and cell viability was measured by Resazurin assay. Viability is shown relative to uninfected DMSO-treated control at respective timepoints. One datapoint represents one organoid. Data are presented as a mean ± SD of n = 2 biological replicates. Statistical significance was determined using two-way ANOVA with Dunnett’s multiple comparison analysis. * p < 0.05, ** p < 0.01, ( C ) ZIKV-infected and uninfected whole organoids treated with TH6744 or DMSO for 7 or 10 dpi, stained by Hoechst and imaged by high-content confocal microscopy. Representative images of n = 2 biological replicates per condition. The scale bar equals 500 μm. Hoechst in blue. ( D ) ZIKV-infected organoids that were treated with TH6744 or DMSO were fixed at 7 dpi, cryosectioned and stained with Hoechst and ZIKV NS1 protein antibody. ZIKV NS1 intensity was quantified using Cellprofiler software. Data are presented as single values and mean ± SD from n = 2 biological replicates. Statistical significance was determined using one-way ANOVA with Dunnett’s multiple comparison analysis. * p < 0.05, ** p < 0.01. ( E ) ZIKV-infected organoids treated with TH6744 or DMSO were fixed at 7 dpi, cryosectioned, stained with Hoechst and ZIKV NS1 protein antibody and imaged by confocal microscopy. Representative images of n = 2 biological replicates per condition. Scale bar equals 500 μm in overview image and 100 μm in close-up. ZIKV NS1 protein in red, Hoechst in blue. ( F ) Viral titer from ZIKV-infected and compound treated organoids at indicated time points was determined by an end-point dilution assay. Viral titer is presented as relative to ZIKV-infected DMSO-treated control at respective timepoints. Data are presented as a mean ± SD from n = 2 biological replicates. Statistical significance was determined using two-way ANOVA with Sidak’s multiple comparison analysis. * p < 0.05, ** p < 0.01.

Article Snippet: For supernatants, the obtained Ct values were converted to the number of ZIKV RNA molecules using a standard curve that was created using Quantitative Synthetic ZIKV RNA from ATCC (ATCC ® VR-3252SDTM).

Techniques: Infection, Resazurin Assay, Control, Comparison, Staining, Confocal Microscopy, Software, End-point Dilution Assay

Journal: Viruses

Article Title: Broadly Active Antiviral Compounds Disturb Zika Virus Progeny Release Rescuing Virus-Induced Toxicity in Brain Organoids

doi: 10.3390/v13010037

Figure Lengend Snippet: TH6744 disturbs late replication cycle steps in ZIKV life cycle. ( A – D ) U87 cells were infected with ZIKV (MOI 1) and treated with indicated compounds for 24, 48 or 72 h. ( A ) ZIKV RNA levels inside the cells were quantified by qPCR. ( B ) Number of ZIKV-infected cells was quantified by virus infectivity assay. ( C ) ZIKV RNA levels in the supernatant were quantified by one-step qRT-PCR. ( D ) ZIKV titers were quantified by end-point dilution assay. ( E ) Schematic overview of time-of-addition experimental setup in U87 cells. ( F ) U87 cells were infected with ZIKV (MOI 10) for a total of 24 h and treated with DMSO control, 10 μM TH6744 or 100 μM Ribavirin for indicated periods during early (entry), late (budding) or throughout replication cycle (post-inoculation). ZIKV titers were quantified by end-point dilution assay. ( G ) Schematic overview of budding experimental setup. ( H ) U87 cells were infected with ZIKV (MOI 10) for 22 h and treated with indicated TH6744 doses for 2 h. Intracellular ZIKV particles were obtained by mechanical cell lysis and extracellular ZIKV particles from the supernatants and both were quantified by end-point dilution assay. ( I ) Average fractions of intracellular and extracellular infectious ZIKV particles of the total particles quantified in ( H ). ( A – D , F , H ) Data are expressed as a mean ± SD from at least n = 3 biological replicates. Statistical significance was determined by using one-way ANOVA with Dunnett’s multiple comparison analysis. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For supernatants, the obtained Ct values were converted to the number of ZIKV RNA molecules using a standard curve that was created using Quantitative Synthetic ZIKV RNA from ATCC (ATCC ® VR-3252SDTM).

Techniques: Infection, Virus, Quantitative RT-PCR, End-point Dilution Assay, Control, Lysis, Comparison

Journal: Nutrients

Article Title: Novel Therapeutic Nutrients Molecules That Protect against Zika Virus Infection with a Special Note on Palmitoleate

doi: 10.3390/nu15010124

Figure Lengend Snippet: List of novel nutrient molecules and mechanism of protection against ZIKV infection.

Article Snippet: Palmatine , ATCC VR-1843 , Vero cells , Prevents ZIKV binding and entry , Virucidal effects , [ ] .

Techniques: Infection, Activity Assay, Derivative Assay, Binding Assay, Inhibition, Glycoproteomics, Virus, Isolation, In Vitro, In Vivo, Blocking Assay, Membrane, Recombinant, Expressing, Concentration Assay

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: Schematic overview of the anti-SARS-CoV-2 drug screening procedure. ( a ) ACE2 targeted inhibitors and compounds from the natural product library were applied prior to infection with SARS-CoV-2 (pre-infection treatment) while compounds from the FDA-approved library and the flavonoid library were applied post infection (post- infection treatment). Out of 2191 compounds tested, 121 displayed viral CPE reductions and 7 were selected for further validations using Vero E6 and HuH7 cell lines and hNEC. ( b ) Uninfected cells, infected cells treated with 0.1% DMSO, and infected cells treated with remdesivir were included as controls in the screen. Following treatment with citicoline, pravastatin sodium, tenofovir alafenamide, imatinib mesylate, calcitriol, dexlansoprazole, and prochlorperazine dimaleate, reduced CPE was observed when compared to the DMSO control.

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: Drug discovery, Infection, Control

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: Summary of compounds identified from the primary screen that exhibit potential antiviral effects against SARS-CoV-2 infection.

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: Infection, Reverse Transcription, Reflux

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: Validation of primary hits. For validating compounds with activity pre-infection, Vero E6 cells were first pre-treated with increasing concentrations of ( a ) citicoline, ( b ) pravastatin sodium and ( c ) tenofovir alafenamide prior to infection with SARS-CoV-2. Similarly, Huh7 cells were pre-treated with ( d ) citicoline, ( e ) pravastatin sodium and ( f ) tenofovir alafenamide and subsequently infected with SARS-CoV-2. For post-infection treatment validation, Vero E6 cells were infected with SARS-CoV-2 and treated with increasing concentrations of ( g ) imatinib mesylate, ( h ) calcitriol, ( i ) dexlansoprazole and ( j ) prochlorperazine dimaleate. Similarly, HuH7 cells were also infected then treated with a range of concentrations of ( k ) imatinib mesylate, ( l ) calcitriol, ( m ) dexlansoprazole and ( n ) prochlorperazine dimaleate. The primary and secondary exes correspond to the viral titre and relative cell viability, and the dashed line represents the CC 50 cut-off for cell viability. One-way ANOVA and Dunnett’s post-test were used to determine statistical differences, with * denoting p < 0.05, ** denoting p < 0.01 and *** denoting p < 0.001. Error bars represent the standard deviation observed from the means of triplicates performed for both cell viability and dose-dependent inhibition studies.

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: Biomarker Discovery, Activity Assay, Infection, Standard Deviation, Inhibition

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: Validation of primary hits in hNECs. Selected primary hits were further validated in hNECs. Cells were either pre-treated with 10 μM citicoline prior to SARS-CoV-2 infection or treated following SARS-CoV-2 infection with 10 μM imatinib mesylate and 10 μM calcitriol. One-way ANOVA and Dunnett’s post-test were used to determine statistical differences, with ** denoting that p < 0.01. Error bars represent the standard deviation observed from the means of triplicates performed for dose-dependent inhibition studies.

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: Biomarker Discovery, Infection, Standard Deviation, Inhibition

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: SARS-CoV-2 and calcitriol modulate vitamin D Receptor (VDR), 24-hydroxylase (24(OH)ase), and cathelicidin (LL-37) in Vero E6 and Huh7 cells. In the first part of the experiment, Vero E6 and Huh7 cells were either mock-infected with media, or SARS-CoV-2 infected and harvested at indicated timepoints in the absence of calcitriol for baseline VDR, 24(OH)ase, and LL-37 mRNA expression. Vero E6 expression of ( a ) VDR, ( b ) 24(OH)ase and ( c ) LL-37 and Huh7 expression of ( d ) VDR, ( e ) 24(OH)ase and ( f ) LL-37 were as shown. In the second part of the experiment, Vero E6 and Huh7 cells were either mock-infected with media or SARS-CoV-2 for 1 h as above, then treated with 0.01, 0.1, 1, 10 or 20 µM calcitriol following infection. At 48 h post infection, the cells were harvested to check for VDR, 24(OH)ase, and LL-37 mRNA expression. Vero E6 mRNA expression of ( g ) VDR, ( i ) 24(OH)ase and ( k ) LL-37 and Huh7 expression of ( h ) VDR, ( j ) 24(OH)ase and ( l ) LL-37 were as shown. Data are represented as relative quantity to 0.1% DMSO control, or as relative quantity to mock-infected control. One-way ANOVA and Dunnett’s post-test and paired t -test were used to determine statistical differences, with * denoting p < 0.05, ** denoting p < 0.01 and *** denoting p < 0.001. Error bars represent the standard deviation observed from the means of triplicates performed for mRNA expression studies.

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: Infection, Expressing, Control, Standard Deviation

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: Western blot analysis of SARS-CoV-2 spike and VDR proteins in Vero E6 cells. ( a ) Vero E6 cells are either mock infected with media (−) or with SARS-CoV-2 (+) for 1 h. After 1 h post-infection, the cells are then either untreated (−) or treated (+) with 20 µM calcitriol for 6-, 24- and 48- hours before harvesting of total cell lysate. ( b ) Densitometry quantitation of protein expression levels is shown as fold changes to beta-actin. Full-length blots are shown in .

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: Western Blot, Infection, Quantitation Assay, Expressing

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: In vivo study of calcitriol treatment. 8-week old K18-hACE2 male and female mice were separated into treatment and non-treatment groups. Treatment study group has been given 3 days pre-treatment and 5 days post-treatment of calcitriol (5 μg/kg) via intraperitoneal injection. The non-treatment group has been given a PBS as a treatment control. Next, 10 3 PFU of SARS-CoV-2 (L, Alpha, and Beta variants) were inoculated through intranasal delivery to all groups after pre-treatment. Physiological parameters: body weight ( a – c ), survival rate ( d – f ), and physiological conditions were monitored after infections. For comparison between individual physiological conditions, 5 dpi infected mice were compared between treatment groups of all three variant infected mice ( g – i ). Scoring of physiological conditions was based on five criteria: appearance of the mouse coat, level of consciousness, activity level, eye condition, and respiratory quality. To compare the severity of damage in mouse tissues after infection, left lung lobes from 4 dpi mice were processed for histological analyses and scored based on six criteria to determine severity ( j – l ). The six criteria are inflammatory cell infiltration, haemorrhage, oedema, bronchial epithelial cell damage, degeneration of alveolar epithelial cells, and parenchymal wall expansion. For viral load determination, right lung, brain, liver, and spleen tissues from the same 4 dpi mice were harvested and homogenised for virus titration ( m – o ). Data is not shown for liver and spleen tissues. Statistical significance is determined with a two-tailed unpaired t -test, with * denoting p < 0.05. Survival group: variant L: n = 6 (treatment and no treatment); Alpha: n = 6 (treatment), n = 4 (no treatment); Beta: n = 6 (treatment), n = 5 (no treatment). 4 dpi group: variant L: n = 7 (treatment and no treatment); Alpha and Beta: n = 5 each (treatment and no treatment). Mock infection, n = 3.

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: In Vivo, Injection, Control, Comparison, Infection, Variant Assay, Activity Assay, Virus, Titration, Two Tailed Test

Journal: Scientific reports

Article Title: Development of a label-free electrochemical aptasensor for Rift Valley fever virus detection.

doi: 10.1038/s41598-024-74314-7

Figure Lengend Snippet: Fig. 6. Comparative Analysis of the RVFV Aptasensor’s Response to 50 ng/mL RVFV, ZIKA, Dengue, Rubella, BSA, and Globulin, with Error Bars Indicating Standard Deviation from Three Replicate Measurements.

Article Snippet: RVFV Nucleoprotein, Rubella virus-like particles, Zika Virus Envelope Protein and Dengue Virus NS1 Protein Serotypes 1–4 was obtained from The Native Antigen Company (Kidlington, United Kingdom), Pure Cube Ni-NTA (Nickel-Nitrilotriacetic acid) Agarose beads were purchased from Cube Biotech (Monheim, Germany).

Techniques: Standard Deviation

Journal: Emerging microbes & infections

Article Title: Extracellular acidosis enhances Zika virus infection both in human cells and ex-vivo tissue cultures from female reproductive tract.

doi: 10.1080/22221751.2021.1932606

Figure Lengend Snippet: Figure 2. Acidosis enhances ZIKV infection in cells derived from the female reproductive tract and other target cells. Cells were incubated with ZIKV (A1C1-V2) at different pH values for 1 h at 37°C. Then, cells were washed and infection was assessed 72 h later. Infection of genital tract-derived cells VK2/E6E7 (A), Ect1/E6E7 (B), and End1/E6E7 (C) was quantified by measuring the extra- cellular concentration of NS-1 by Luminex. Infection of human endometrial fibroblasts (T-HESC) was quantified by qPCR (D). Infec- tion of monocyte-derived macrophages (MDM) was quantified by measuring the extracellular concentration of NS-1 by Luminex (E). Primary culture-derived human astrocytes (NHA) infection was measured by qPCR (F). Human ectocervical (G) and endocervical (H) tissue cultures were dissected into 2 mm [3] pieces and infected with ZIKV (VR-1844 moi 0.1) either at pH 7.3 or 6.2 for 1 h at 37°C. The viral production was evaluated at days 3, 6, 10, 13, 17, and 21 by measuring ZIKV NS1 in the culture supernatant. Results in A, B, C, D, and F represent data of two independent experiments. Data in E comprises the results of four different donors. Panels H and G were constructed with data of three independent experiments per explant type, and data were analysed using paired two-way ANOVA. Results are presented as mean ± SEM.

Article Snippet: The standard curve was obtained by serial dilution of recombinant NS1 protein (Fitzgerald, cat no. 30-1931).

Techniques: Infection, Derivative Assay, Incubation, Concentration Assay, Luminex, Construct