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

Journal: PLoS ONE

Article Title: Zika virus as an oncolytic treatment of human neuroblastoma cells requires CD24

doi: 10.1371/journal.pone.0200358

Figure Lengend Snippet: A) Western blot analysis of Zika virus infections in neuroblastoma cells. Total protein analysis was performed for Zika Envelope protein and NS1 (Non-Structural 1) protein compared to GAPDH control. The neuroblastoma cell lines tested included IMR-32, SMS-KAN, SK-N-AS, LA-N-6, SK-N-Be(1), and CHLA-42, using Vero cells as an infection control. All cells were compared to control cells treated with non-infected conditioned media. Samples were assessed 3 days after infection (MOI = 10). These results were representative of the combined data of experiments performed in triplicate. B) Viral Titer (TCID50) assays of IMR-32 and SK-N-AS cells at Day 2 and 3 post-infection. Data is composed of three biological replicates examined in sextuplicate, with error bars representing standard deviation. ** p > 0.05, Student’s t-test. C) Immunofluorescence labeling of Zika viral Envelope protein in IMR-32 and SK-N-AS cells at Day 3 post-infection. Envelope staining is in red (Alexa Fluor 647) and nuclei are stained in blue (DAPI). Samples are also shown together (merged). Cells were scanned using a Nikon A1R VAAS laser point- and resonant-scanning confocal microscope. Images are at a magnification of 40x with a 4x zoom.

Article Snippet: Proteins were transferred to nitrocellulose membranes (0.2 um, BioRad, Cat# 1620112) and probed with the following primary antibodies: Anti-Zika virus NS1 (non-structural protein 1) (One World Lab, Cat# 55964) at 1/200, anti-Zika virus Envelope (Env) (GeneTex, Cat# GTX133314) at 1/1000, and anti-GAPDH (Santa Cruz, FL-335) at 1/2000.

Techniques: Western Blot, Virus, Control, Infection, Standard Deviation, Immunofluorescence, Labeling, Staining, Microscopy

Journal: PLoS ONE

Article Title: Zika virus as an oncolytic treatment of human neuroblastoma cells requires CD24

doi: 10.1371/journal.pone.0200358

Figure Lengend Snippet: A) Western blot analysis of siRNA-mediated knock-down of CD24 expression in IMR-32 cells. Samples include Negative Control siRNA and CD24 siRNA. B) Western blot analysis of the expression of Envelope protein and NS1 (Non-Structural 1) protein in IMR-32 cells after siRNA-mediated knock-down of CD24 expression, 96 hours after Zika infection (MOI = 10). Samples include control cells treated with non-infected conditioned media and infected IMR-32 cells transfected with either Negative Control siRNA or CD24 siRNA. C) Western blot analysis of CD24 expression in the human neuroblastoma cell line SK-N-AS, comparing wild type (WT) to stably selected “Vector Only” (VO), CD24 variant 1 (V1), and CD24 variant 7 (V7). D) Western blot analysis of Zika NS1 protein expression 96 hours after Zika infection in CD24-stably expressing SK-N-AS cells, comparing wild type (WT) to stably selected Vector Only (VO), CD24 variant 1 (V1), and CD24 variant 7 (V7). GAPDH was used as a load control for all experiments. All results are representative of the combined data of experiments performed in triplicate.

Article Snippet: Proteins were transferred to nitrocellulose membranes (0.2 um, BioRad, Cat# 1620112) and probed with the following primary antibodies: Anti-Zika virus NS1 (non-structural protein 1) (One World Lab, Cat# 55964) at 1/200, anti-Zika virus Envelope (Env) (GeneTex, Cat# GTX133314) at 1/1000, and anti-GAPDH (Santa Cruz, FL-335) at 1/2000.

Techniques: Western Blot, Knockdown, Expressing, Negative Control, Infection, Control, Transfection, Stable Transfection, Plasmid Preparation, Variant Assay

Journal: The Journal of Pathology: Clinical Research

Article Title: Optimization of commercially available Zika virus antibodies for use in a laboratory‐developed immunohistochemical assay

doi: 10.1002/cjp2.84

Figure Lengend Snippet: A comparison of immunostaining between commercially available ZIKV antibodies in ZIKV‐infected Vero cells (A–F) and uninfected Vero cells (G–L). Objective magnification, 40X. A high resolution version of this figure is available in supplementary material, Figure S1.

Article Snippet: GTX‐C , Zika virus NS1 protein , GeneTex , GTX13307 , Rabbit, polyclonal , 1:100 PK 1:100 ARE , 1:500 PK 1:500 ARE , 1:1000 PK 1:1000 ARE , 1:500 ARE.

Techniques: Comparison, Immunostaining, Infection

Journal: The Journal of Pathology: Clinical Research

Article Title: Optimization of commercially available Zika virus antibodies for use in a laboratory‐developed immunohistochemical assay

doi: 10.1002/cjp2.84

Figure Lengend Snippet: A comparison of immunostaining between the commercially available ZIKV antibodies using ZIKV‐infected case control (A–F) and uninfected CNS tissue (G–L). Objective magnification, 20X. A high resolution version of this figure is available in supplementary material, Figure S2.

Article Snippet: GTX‐C , Zika virus NS1 protein , GeneTex , GTX13307 , Rabbit, polyclonal , 1:100 PK 1:100 ARE , 1:500 PK 1:500 ARE , 1:1000 PK 1:1000 ARE , 1:500 ARE.

Techniques: Comparison, Immunostaining, Infection, Control

Journal: The Journal of Pathology: Clinical Research

Article Title: Optimization of commercially available Zika virus antibodies for use in a laboratory‐developed immunohistochemical assay

doi: 10.1002/cjp2.84

Figure Lengend Snippet: Commercially available antibodies optimized for IHC: antibody source, characteristics, and pretreatment conditions

Article Snippet: GTX‐C , Zika virus NS1 protein , GeneTex , GTX13307 , Rabbit, polyclonal , 1:100 PK 1:100 ARE , 1:500 PK 1:500 ARE , 1:1000 PK 1:1000 ARE , 1:500 ARE.

Techniques: Virus

Journal: The Journal of Pathology: Clinical Research

Article Title: Optimization of commercially available Zika virus antibodies for use in a laboratory‐developed immunohistochemical assay

doi: 10.1002/cjp2.84

Figure Lengend Snippet: Cross‐reactivity profile of commercially available ZIKV antibodies tested by IHC against other flaviviruses and chikungunya virus (sg: serogroup; neg: negative; pos: positive)

Article Snippet: GTX‐C , Zika virus NS1 protein , GeneTex , GTX13307 , Rabbit, polyclonal , 1:100 PK 1:100 ARE , 1:500 PK 1:500 ARE , 1:1000 PK 1:1000 ARE , 1:500 ARE.

Techniques: Virus