zika mr766 atcc vr 84 mw143022 1 uganda strain  (ATCC)

Journal: ACS Omega

Article Title: Evolutionary Dynamics-Aware De Novo Design of a Thermostable NS1-Derived Diagnostic Protein for Yellow Fever Virus

doi: 10.1021/acsomega.6c07158

Figure Lengend Snippet: Maximum clade credibility (MCC) trees, population dynamics, and temporal conservation mapping of the YFV NS1 protein. (a) The MCC tree for the YFV polyprotein is shown together with the SkyGrid reconstruction of effective population size over time. The SkyGrid plot (white to light red gradient) displays the estimated median and corresponding 95% HPD interval. Nodes with posterior support ≥80% are indicated by circles. (b) The structure of the NS1 monomer, in which residues inferred to have changed throughout YFV evolutionary history are highlighted in red. NS1 coordinates within the viral polyprotein sequence are shown in parentheses.

Article Snippet: Then, 96 well microplates were coated with 100 μL per well of purified ZIKV NS1 (The Native Antigen Company, Kidlington, UK), recombinant YFV NS1 (R&D Systems, Minneapolis, USA), or YFV-Scaffold at 4 μg/mL and incubated overnight at 4 °C.

Techniques: Sequencing

Journal: ACS Omega

Article Title: Evolutionary Dynamics-Aware De Novo Design of a Thermostable NS1-Derived Diagnostic Protein for Yellow Fever Virus

doi: 10.1021/acsomega.6c07158

Figure Lengend Snippet: Top-ranked designed protein containing the selected motif from the YFV NS1 protein. (a) Scatter plot showing the distribution of radius of gyration (RG) versus number of backbone contacts (NC), both normalized by protein length, for each backbone generated by RFdiffusion. (b) Predicted structure of the designed protein colored according to the AlphaFold 3 pLDDT confidence score. (c) Superimposition of the designed protein (white) and its RFdiffusion-generated backbone (green). (d) Superimposition of the predicted protein structure and its native epitope (orange).

Article Snippet: Then, 96 well microplates were coated with 100 μL per well of purified ZIKV NS1 (The Native Antigen Company, Kidlington, UK), recombinant YFV NS1 (R&D Systems, Minneapolis, USA), or YFV-Scaffold at 4 μg/mL and incubated overnight at 4 °C.

Techniques: Generated

Journal: ACS Omega

Article Title: Evolutionary Dynamics-Aware De Novo Design of a Thermostable NS1-Derived Diagnostic Protein for Yellow Fever Virus

doi: 10.1021/acsomega.6c07158

Figure Lengend Snippet: Comparison of the structural properties of the epitope in its native conformation and within the designed YFV NS1 protein. (a) Relative solvent-accessible surface area (SASA) per residue for the native epitope (blue) and the designed protein (orange). (b) Number of backbone contacts per residue, defined as the number of atoms from adjacent residues within 4 Å, using the same color scheme as in (a). (c) Electrostatic potential surface of the epitope in the native (right) and designed (left) protein contexts. Electrostatic potential ranges from −5 kT/e (red) to +5 kT/e (blue) and is mapped onto the van der Waals surface.

Article Snippet: Then, 96 well microplates were coated with 100 μL per well of purified ZIKV NS1 (The Native Antigen Company, Kidlington, UK), recombinant YFV NS1 (R&D Systems, Minneapolis, USA), or YFV-Scaffold at 4 μg/mL and incubated overnight at 4 °C.

Techniques: Comparison, Solvent, Residue

Journal: bioRxiv

Article Title: Structure informed xrRNA2 mutations in Zika virus provide a roadmap for vaccine development

doi: 10.64898/2026.05.27.728262

Figure Lengend Snippet: Zika virus xrRNA2 exhibits increased resistance to mutagenesis compared to xrRNA1. ( A ) Diagram of ZIKV 3’ UTR showing crystal-structure derived predictions of xrRNA2 L1 interactions with the 5’ end of the structure (Blue). ( B ) Diagram of mutation locations in ZIKV xrRNA1 (C104415G) and ZIKV xrRNA2 (C10496G) predicted to disrupt tertiary structure interaction with the phosphate backbone of the pseudoknot and allow for Xrn1 degradation. ( C ) Representative northern blot (n=2) of RNA from Vero cells following infection with WT ZIKV, ZIKV xrRNA1 (C10415G) in L1, and ZIKV xrRNA2 (C10496G) in L1 (MOI 1, 48 hours post-infection). 3’UTR radiolabeled to detect sfRNA production.

Article Snippet: Zika Virus Total Antibody Detection Assay Development Kit from Native Antigen company (Cat # ELS61258) was purchased and used to determine total antibody to ZIKV.

Techniques: Virus, Mutagenesis, Derivative Assay, Northern Blot, Infection

Journal: bioRxiv

Article Title: Structure informed xrRNA2 mutations in Zika virus provide a roadmap for vaccine development

doi: 10.64898/2026.05.27.728262

Figure Lengend Snippet: Disruption of Zika virus xrRNA2 L1 (U10510A) with the 5’ end results in a small plaque phenotype. We produced clone-derived ZIKV virus isolates with ( A ) WT ZIKV xrRNA2, ( B ) ZIKV xrRNA2 mutations at C10518G, U10510A, and ( C ) ZIKV xrRNA2 U10519A. ZIKV xrRNA2 (C10518G, U10510A) reverted to ZIKV xrRNA2 U10510A. Vero cells were inoculated with WT ZIKV or ZIKV xrRNA2 U10510A (referred to as ZIKV L1U,A) at a MOI of 1.0. There was no significant difference in ZIKV genome copies from ( D ) supernatant or ( E ) cell lysates comparing WT ZIKV and ZIKV L1U,A. ( F ) Infectious plaque forming units (PFU) exhibited no significant difference in supernatant from WT ZIKV and ZIKV L1U,A inoculated vero cells. ( G,H ) ZIKV plaques from ZIKV L1U,A exhibited significantly smaller plaque size compared to WT ZIKV plaques. *p<0.05, **p<0.01, Student t-test.

Article Snippet: Zika Virus Total Antibody Detection Assay Development Kit from Native Antigen company (Cat # ELS61258) was purchased and used to determine total antibody to ZIKV.

Techniques: Disruption, Virus, Produced, Derivative Assay

Journal: bioRxiv

Article Title: Structure informed xrRNA2 mutations in Zika virus provide a roadmap for vaccine development

doi: 10.64898/2026.05.27.728262

Figure Lengend Snippet: Targeted nucleotide mutations in ZIKV xrRNA2 eliminate sfRNA2 production. We produced clone-derived ZIKV virus isolates with ( A ) WT ZIKV xrRNA2, ( B ) ZIKV xrRNA2 mutations at C10496G, C10518G, and U10510A, termed X2.L1 ZIKV. ( C,D ) X2.L1 ZIKV exhibited stable sequencing with no reversion, ZIKV genome copy number, and ZIKV infectious particles that were slightly decreased compared to WT ZIKV. *p<0.05, t-test. ( E ) Gel image of a representative in vitro Xrn1 RNA degradation assay of WT ZIKV 3’UTR compared to X2.L1 ZIKV 3’UTR showing loss of sfRNA2 with X2.L1 mutations in the xrRNA2 structure (n=2).

Article Snippet: Zika Virus Total Antibody Detection Assay Development Kit from Native Antigen company (Cat # ELS61258) was purchased and used to determine total antibody to ZIKV.

Techniques: Produced, Derivative Assay, Virus, Sequencing, In Vitro, Degradation Assay

Journal: bioRxiv

Article Title: Structure informed xrRNA2 mutations in Zika virus provide a roadmap for vaccine development

doi: 10.64898/2026.05.27.728262

Figure Lengend Snippet: X2.L1 ZIKV exhibits attenuation in cell culture. Vero cells were inoculated with WT ZIKV or X2.L1 ZIKV at a MOI of 0.1 followed by analysis at indicated time points for ( A ) ZIKV genome copies in the supernatant, ( B ) ZIKV genome copies in cell lysates, and ( C ) infectious ZIKV particles in the supernatant as measured by focus forming units (FFU). *p=0.001 two-way ANOVA. ( D ) Vero cells were inoculated with WT ZIKV or X2.L1 ZIKV (MOI=1) and cell injury was measured at the indicated time points using an XTT assay. *p<0.001, two-way ANOVA. ( E ) Vero cells were inoculated with WT ZIKV or X2.L1 ZIKV (MOI=1) and treated with serial dilutions of interferon-alpha2 followed by determination of genome copies at 48 hours post-infection. Using the dilution curve of interferon-alpha2, we determined the concentration of interferon required to inhibit 50% ZIKV genomes (IC50). X2.L1 ZIKV exhibited an IC50=30.9 and WT ZIKV exhibited an IC50=91.44. N=10, Least squares regression.

Article Snippet: Zika Virus Total Antibody Detection Assay Development Kit from Native Antigen company (Cat # ELS61258) was purchased and used to determine total antibody to ZIKV.

Techniques: Cell Culture, XTT Assay, Infection, Concentration Assay

Journal: bioRxiv

Article Title: Structure informed xrRNA2 mutations in Zika virus provide a roadmap for vaccine development

doi: 10.64898/2026.05.27.728262

Figure Lengend Snippet: X2.L1 ZIKV is Attenuated in Immune Compromised Mice Type I interferon receptor knockout (IFNAR1 KO) mice were inoculated with mock control, WT ZIKV, or X2.L1 ZIKV (10 4 pfu, intraperitoneal [ip] inoculation), and mice were euthanized at day 8 post-infection. ( A ) There was no significant difference in weight changes between mock, WT ZIKV, and X2.L1 ZIKV inoculated mice. At day 8 post-infection, there was no significant difference in ZIKV genome copy number between mock, WT ZIKV, and X2.L1 ZIKV in the ( B ) brain or ( C ) spleen tissue (n=6, *p>0.05(n.s.), one-way ANOVA). Next, we inoculated type I and type II interferon receptor knockout (AG129) mice with mock control, WT ZIKV, or X2.L1 ZIKV (10 4 pfu, intraperitoneal [ip] inoculation), and mice were euthanized at day 8 post-infection. ( D ) AG129 mice inoculated with WT-ZIKV exhibited 10-20% increased weight loss compared to mock and X2.L1 ZIKV inoculated mice (*p<0.0001, n=18, two-way ANOVA, repeated measures). At day 8 post-infection, ( E ) there was no difference in ZIKV genome copy number in the spleens of WT ZIKV and X2.L1 ZIKV inoculated AG129 mice; however, ( F ) AG129 mice inoculated with X2.L1 ZIKV exhibited a 90% decrease in mean ZIKV genome copies in brain tissue compared to WT ZIKV inoculated mice (n=6, *p<0.0001, one-way ANOVA with multiple comparisons). Next, we inoculated IFNAR KO mice with Zika Paraiba 01 Brazil 2015 (WT ZIKV Bazil) compared to X2.L1 ZIKV (10 4 pfu, ip inoculation). We found that WT ZIKV Brazil inoculated mice exhibited significantly increased ( G ) weight loss (n=6, *p=0.0014, two-way ANOVA, repeated measures) compared to X2.L1 inoculated mice. (H) Additionally, X2.L1 ZIKV inoculated mice exhibited significantly decreased mortality (*p=0.0179, Logrank test) compared to WT ZIKV Brazil inoculated IFNAR KO mice.

Article Snippet: Zika Virus Total Antibody Detection Assay Development Kit from Native Antigen company (Cat # ELS61258) was purchased and used to determine total antibody to ZIKV.

Techniques: Knock-Out, Control, Infection

Journal: bioRxiv

Article Title: Structure informed xrRNA2 mutations in Zika virus provide a roadmap for vaccine development

doi: 10.64898/2026.05.27.728262

Figure Lengend Snippet: X2.L1 ZIKV Inoculation produces protective neutralizing antibodies IFNAR KO mice were inoculated with mock, WT ZIKV, or X2.L1 ZIKV (10 4 pfu, ip) and euthanized at day 21 post-infection for serum collection. ( A ) Total anti-ZIKV antibody were significantly increased (*p<0.0001, one-way ANOVA, multiple comparisons) in WT ZIKV inoculated mice and there was a non-statistically significant increase in X2.L1 ZIKV inoculated mice compared to mock-inoculated mice. ( B ) Neutralization antibody titers were determined in the same experimental groups using focus-forming unit neutralization 50 (FRNT50) assays showing that both WT-ZIKV (FRNT=1:180,000) and X2.L1 ZIKV (FRNT=1:6,024) exhibited high titer neutralization in the serum. ( C ) Next, serum from mock inoculated and X2.L1 inoculated mice was transferred to AG129 mice (100microL of serum ip) and mice were inoculated with WT ZIKV Brazil after 24 hours. ( D ) AG129 mice inoculated with X2.L1 ZIKV serum exhibited protection from weight loss following lethal WT ZIKV Brazil challenge (10 4 pfu, subcutaneous inoculation) compared to AG129 mice inoculated with serum from infected serum and ( E ) X2.L1 ZIKV serum protected naïve AG129 mice from WT ZIKV Brazil mortality (n=6, *p=0.0009, Logrank test).

Article Snippet: Zika Virus Total Antibody Detection Assay Development Kit from Native Antigen company (Cat # ELS61258) was purchased and used to determine total antibody to ZIKV.

Techniques: Infection, Neutralization

Journal: bioRxiv

Article Title: Structure informed xrRNA2 mutations in Zika virus provide a roadmap for vaccine development

doi: 10.64898/2026.05.27.728262

Figure Lengend Snippet: Vaccination with X2.L1 ZIKV protect Immunocompromised mice from ZIKV challenge. ( A ) IFNAR1 KO mice were inoculated with mock or X2.L1 ZIKV (10 4 pfu, ip) followed by WT ZIKV Brazil (10 4 pfu, sc) challenge at day 21 post-vaccination. ( B ) X2.L1 ZIKV vaccinated mice exhibited no significant weight loss following WT ZIKV Brazil challenge compared to mock vaccinated animals (n=12, *p<0.0001, two-way ANOVA with multiple comparisons). ( C ) IFNAR KO mice inoculated with mock vaccination exhibited a median survival of 11.5 days compared to 0% mortality in X2.L1 ZIKV vaccinated mice following WT ZIKV Brazil challenge (n=12, *p=0.0018, Logrank test). At 8 days post-infection, mice vaccinated with mock or X2.L1 ZIKV and challenged with WT ZIKV Brazil were euthanized for ZIKV genome copy analysis in the spleen and brain tissue. ( D ) X2.L1 ZIKV vaccinated mice exhibited significantly decreased ZIKV genome copies in the spleen (n=6, *p=0.0234, t-test) and ( E ) in brain tissue (n=6, *p=0.037, t-test).

Article Snippet: Zika Virus Total Antibody Detection Assay Development Kit from Native Antigen company (Cat # ELS61258) was purchased and used to determine total antibody to ZIKV.

Techniques: Infection

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Aedes aegypti salivary gland extract enhances Zika virus replication through immune modulation

doi: 10.1590/0074-02760250272

Figure Lengend Snippet: Aedes aegypti salivary gland extract (SGE) enhances Zika virus (ZIKV) replication in Vero cells. (A) Dose-response curve showing plaque-forming units per millilitre (PFU/mL) recovered from Vero cells infected with ZIKV [multiplicity of infection (MOI) = 0.1] in the presence of increasing concentrations of SGE (0.5, 1, 2.5, and 5 µg/mL). Data are expressed as mean ± standard error of the mean (SEM) from three independent experiments (n = 3). Statistical significance was determined by one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test. Asterisks indicate statistical significance compared with ZIKV alone (p < 0.05). (B) Representative plaque assay images of Vero cell monolayers infected with ZIKV (MOI 0.1) and treated with SGE at the indicated concentrations, illustrating the dose-dependent increase in cytopathic effects.

Article Snippet: Quantification of ZIKV RNA was performed using the AllplexTM Zika Virus Assay (Seegene®), a multiplex Real-Time PCR kit designed for specific detection of ZIKV RNA.

Techniques: Virus, Infection, Plaque Assay

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Aedes aegypti salivary gland extract enhances Zika virus replication through immune modulation

doi: 10.1590/0074-02760250272

Figure Lengend Snippet: leukocyte death profiles in peripheral blood mononuclear cells (PBMCs) following Zika virus (ZIKV) infection and Aedes aegypti salivary gland extract (SGE) exposure. (A) Absolute number of dead leukocytes after 72 h of culture with medium only, SGE (5 µg/mL), ZIKV, or ZIKV+SGE (5 µg/mL). (B) Absolute counts of Annexin V⁺ (early apoptotic), PI⁺ (late apoptotic/necrotic), and Annexin V⁺/PI⁺ (late apoptotic/secondary necrotic) leukocytes across groups. (C) Cell death dynamics representation (Annexin V⁺, PI⁺, Annexin V⁺/PI⁺) illustrating the redistribution of death profiles under different experimental conditions. Data represent PBMCs from five independent donors, each tested in duplicate (individual points shown), with median ± interquartile range (IQR). Statistical comparisons were performed using Friedman test with post-hoc analysis (Panel A: p = 0.0129 overall; ZIKV+SGE vs SGE, p = 0.0109; Panel B: PI⁺ ZIKV+SGE vs SGE, p = 0.0335; Annexin V⁺/PI⁺ ZIKV+SGE vs SGE, p = 0.0327; Annexin V⁺ frequencies, p < 0.0001). Two-way analysis of variance (ANOVA) for the Cell death dynamics (Panel C) detected significant main effects of condition and marker type, and their interaction (p < 0.0001, p = 0.0216, p = 0.0257, respectively).

Article Snippet: Quantification of ZIKV RNA was performed using the AllplexTM Zika Virus Assay (Seegene®), a multiplex Real-Time PCR kit designed for specific detection of ZIKV RNA.

Techniques: Virus, Infection, Marker

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Aedes aegypti salivary gland extract enhances Zika virus replication through immune modulation

doi: 10.1590/0074-02760250272

Figure Lengend Snippet: Zika virus (ZIKV) and salivary gland extract (SGE) co-exposure reduces late apoptotic CD4⁺ T cells while sparing CD8⁺ populations. (A) CD4⁺ T cells: absolute numbers of Annexin V⁺ (early apoptosis), PI⁺ (late apoptosis/necrosis), and Annexin V⁺/PI⁺ (late apoptosis/secondary necrosis) populations after 72 h under the indicated conditions (Media, SGE (5 µg/mL), ZIKV, ZIKV+SGE (5 µg/mL)). A significant difference was detected for the Annexin V⁺/PI⁺ population, with reductions in ZIKV+SGE compared with ZIKV (p = 0.0423). (B) CD8⁺ T cells: no significant differences were observed across groups for any marker in absolute numbers (Friedman test, p > 0.05). Data represent peripheral blood mononuclear cells (PBMCs) from five independent donors shown as individual values with median ± interquartile range (IQR).

Article Snippet: Quantification of ZIKV RNA was performed using the AllplexTM Zika Virus Assay (Seegene®), a multiplex Real-Time PCR kit designed for specific detection of ZIKV RNA.

Techniques: Virus, Marker

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Aedes aegypti salivary gland extract enhances Zika virus replication through immune modulation

doi: 10.1590/0074-02760250272

Figure Lengend Snippet: Zika virus (ZIKV)-induced dendritic cell death is attenuated by Aedes aegypti salivary gland extract (SGE). Absolute numbers of dendritic cells positive for Annexin V⁺ (early apoptosis), PI⁺ (late apoptosis/necrosis), and Annexin V⁺/PI⁺ (late apoptosis/secondary necrosis) were quantified after 72 h of culture under the indicated conditions (Media, SGE (5 µg/mL), ZIKV, ZIKV+SGE (5 µg/mL)). Significant overall differences were observed for early apoptosis (p < 0.0001), late apoptosis/necrosis (p = 0.0087), and late apoptosis/secondary necrosis (p = 0.0023). Post-hoc analyses showed reduced early apoptosis counts in ZIKV+SGE compared to Media (p = 0.0014), fewer late apoptosis/necrosis cells in ZIKV+SGE versus ZIKV (p = 0.0197), and lower late apoptosis/secondary necrosis counts in ZIKV+SGE compared to ZIKV (p = 0.0087). Percentage analyses yielded consistent trends (Annexin V⁺, p < 0.0001; PI⁺, p = 0.0226; Annexin V⁺/PI⁺, p = 0.0167). Data represent peripheral blood mononuclear cells (PBMCs)-derived dendritic cells from five independent donors (no technical replicates), shown as individual values with median ± interquartile range (IQR).

Article Snippet: Quantification of ZIKV RNA was performed using the AllplexTM Zika Virus Assay (Seegene®), a multiplex Real-Time PCR kit designed for specific detection of ZIKV RNA.

Techniques: Virus, Derivative Assay

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Aedes aegypti salivary gland extract enhances Zika virus replication through immune modulation

doi: 10.1590/0074-02760250272

Figure Lengend Snippet: salivary gland extract (SGE) enhances Zika virus (ZIKV) replication in human peripheral blood mononuclear cells (PBMCs). (A) Absolute viral RNA copy numbers in PBMCs cultured under the indicated conditions (Media, SGE, ZIKV, ZIKV+SGE) for 72 h. Each symbol represents one culture replicate; two replicates were performed per donor, resulting in 10 points per group from five independent donors. Quantitative polymerase chain reaction (qPCR) was performed in technical triplicates and averaged per well. (B) Δ viral RNA copies (ZIKV+SGE - ZIKV) calculated for each donor, showing consistent increases across all individuals. (C) Assessment of normality of viral RNA values by Shapiro-Wilk test (p > 0.05), supporting the use of paired analysis. (D) Paired comparison of viral RNA levels between ZIKV and ZIKV+SGE conditions across donors, shown as both absolute copy numbers and log10-transformed values. Wilcoxon matched-pairs test confirmed significantly higher viral loads in ZIKV+SGE (p = 0.002), with a median increase of 1,779 copies per donor (log10 median difference = 0.3367). Data represent PBMCs from five independent donors; each condition was cultured in duplicate, and qPCR performed in technical triplicates. Points depict culture-level replicates, while statistical analyses were performed on donor-level aggregates (duplicates averaged per donor).

Article Snippet: Quantification of ZIKV RNA was performed using the AllplexTM Zika Virus Assay (Seegene®), a multiplex Real-Time PCR kit designed for specific detection of ZIKV RNA.

Techniques: Virus, Cell Culture, Real-time Polymerase Chain Reaction, Comparison, Transformation Assay

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Aedes aegypti salivary gland extract enhances Zika virus replication through immune modulation

doi: 10.1590/0074-02760250272

Figure Lengend Snippet: lipid peroxidation during Zika virus (ZIKV) infection is reduced by Aedes aegypti salivary gland extract (SGE). Thiobarbituric acid reactive substance (TBARS) levels (nmol/mg of protein) were quantified in peripheral blood mononuclear cells (PBMCs) cultures after 72 h under the following conditions: control medium, SGE (1 µg/mL), SGE (5 µg/mL), ZIKV, and ZIKV+SGE (1 µg/mL). ZIKV infection significantly increased TBARS relative to controls (**p < 0.001), while co-treatment with SGE (1 µg/mL) significantly reduced TBARS compared with ZIKV alone (**p < 0.001). Data represent PBMCs from six independent donors (individual values shown) with mean ± standard error of the mean (SEM). Statistical analysis: Kruskal-Wallis with Dunn’s post-hoc test; asterisks denote significant pairwise comparisons as indicated.

Article Snippet: Quantification of ZIKV RNA was performed using the AllplexTM Zika Virus Assay (Seegene®), a multiplex Real-Time PCR kit designed for specific detection of ZIKV RNA.

Techniques: Virus, Infection, Control

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Aedes aegypti salivary gland extract enhances Zika virus replication through immune modulation

doi: 10.1590/0074-02760250272

Figure Lengend Snippet: Aedes aegypti salivary gland extract (SGE) modulates oxidative stress markers in Zika virus (ZIKV)-infected peripheral blood mononuclear cells (PBMCs). (A) Total glutathione (GSH) content, (B) advanced oxidised protein products (AOPP), and (C) nitric oxide (NO) levels in PBMC cultures after 72 h under the indicated conditions (Medium, SGE 1 µg/mL, ZIKV, ZIKV+SGE 1 µg/mL, ZIKV+SGE 5 µg/mL). ZIKV infection increased AOPP and decreased GSH compared with controls, consistent with oxidative imbalance. Co-treatment with SGE reduced AOPP levels and restored GSH content. NO production was significantly higher in ZIKV+SGE compared with ZIKV alone (*p < 0.01). Data represent PBMCs from five independent donors (individual values shown) with mean ± standard error of the mean (SEM). Statistical analysis: Kruskal-Wallis with Dunn’s post-hoc test; asterisks denote significant pairwise differences.

Article Snippet: Quantification of ZIKV RNA was performed using the AllplexTM Zika Virus Assay (Seegene®), a multiplex Real-Time PCR kit designed for specific detection of ZIKV RNA.

Techniques: Virus, Infection

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Aedes aegypti salivary gland extract enhances Zika virus replication through immune modulation

doi: 10.1590/0074-02760250272

Figure Lengend Snippet: characterisation of CD4⁺ T cell frequency, proliferation, and cytotoxic profile following exposure to Zika virus (ZIKV) and/or Aedes aegypti salivary gland extract (SGE). (A) Frequency (%) and (B) absolute number of CD4⁺ T cells in peripheral blood mononuclear cells (PBMCs) cultures after 72 h. (C) Frequency (%) and (D) absolute number of Ki-67⁺ CD4⁺ T cells. (E) Frequency (%) and (F) absolute number of granzyme B⁺ CD4⁺ T cells. Data were analysed by Friedman test followed by Dunn’s multiple comparisons. *p < 0.05; ns: not significant.

Article Snippet: Quantification of ZIKV RNA was performed using the AllplexTM Zika Virus Assay (Seegene®), a multiplex Real-Time PCR kit designed for specific detection of ZIKV RNA.

Techniques: Virus

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Aedes aegypti salivary gland extract enhances Zika virus replication through immune modulation

doi: 10.1590/0074-02760250272

Figure Lengend Snippet: modulation of CD8⁺ T cell frequency, absolute number, proliferation, and cytotoxic profile following exposure to Zika virus (ZIKV) and Aedes aegypti salivary gland extract (SGE). (A) Frequency of CD8⁺ T cells among peripheral blood mononuclear cells (PBMCs) after 72 h of treatment. (B) Absolute number of CD8⁺ T cells in culture. (C) Frequency of granzyme B⁺ CD8⁺ T cells. (D) Absolute number of granzyme B⁺ CD8⁺ T cells. (E) Frequency of Ki-67⁺ CD8⁺ T cells. (F) Absolute number of Ki-67⁺ CD8⁺ T cells. Data are presented as individual donors with medians. Statistical analysis was performed using the Friedman test with Dunn’s multiple comparisons post hoc test. *p < 0.05.

Article Snippet: Quantification of ZIKV RNA was performed using the AllplexTM Zika Virus Assay (Seegene®), a multiplex Real-Time PCR kit designed for specific detection of ZIKV RNA.

Techniques: Virus

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Aedes aegypti salivary gland extract enhances Zika virus replication through immune modulation

doi: 10.1590/0074-02760250272

Figure Lengend Snippet: expansion of dendritic cells and subsets in peripheral blood mononuclear cells (PBMCs) exposed to Zika virus (ZIKV) and Aedes aegypti salivary gland extract (SGE). (A) Absolute numbers of total dendritic cells (DCs) and (B-D) subsets (DC1, mDCs, pDCs) after 72 h of culture under the indicated conditions (Only Media, SGE, ZIKV, ZIKV+SGE). Absolute counts of total DCs, DC1, mDCs, and pDCs were significantly increased in the ZIKV+SGE group compared to controls (p < 0.05). (E) Mean fluorescence intensity (MFI) of CD11c (mDCs), CD141 (DC1), and CD303 (pDCs). No significant differences were observed in CD11c or CD141 expression, while CD303 showed a trend toward reduction under ZIKV+SGE exposure. Data represent PBMCs from five independent donors. Individual values are shown with bars indicating median and interquartile range. Statistical analysis: Friedman test with Dunn’s post-hoc comparisons; p < 0.05 considered significant.

Article Snippet: Quantification of ZIKV RNA was performed using the AllplexTM Zika Virus Assay (Seegene®), a multiplex Real-Time PCR kit designed for specific detection of ZIKV RNA.

Techniques: Virus, Fluorescence, Expressing

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Aedes aegypti salivary gland extract enhances Zika virus replication through immune modulation

doi: 10.1590/0074-02760250272

Figure Lengend Snippet: Aedes aegypti salivary gland extract (SGE) skews cytokine responses toward a Th2 profile during Zika virus (ZIKV) infection. Peripheral blood mononuclear cells (PBMCs) from healthy donors were cultured for 72 h with medium alone (Only Media), SGE, ZIKV, or ZIKV+SGE. (A) Interferon-y (IFN-γ) levels increased after ZIKV infection but were significantly reduced by SGE treatment. (B) Interleukin-4 (IL-4) production was significantly elevated in the presence of SGE, both with and without ZIKV infection. (C) The IFN-γ: IL-4 ratio decreased significantly following SGE treatment, indicating a shift toward a Th2-skewed cytokine profile. Data were analysed using the Friedman test followed by Dunn’s multiple comparisons test; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Quantification of ZIKV RNA was performed using the AllplexTM Zika Virus Assay (Seegene®), a multiplex Real-Time PCR kit designed for specific detection of ZIKV RNA.

Techniques: Virus, Infection, Cell Culture

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Aedes aegypti salivary gland extract enhances Zika virus replication through immune modulation

doi: 10.1590/0074-02760250272

Figure Lengend Snippet: Aedes aegypti salivary gland extract (SGE) modulates cytokine production by antigen-presenting cell lines during Zika virus (ZIKV) infection. JAWS II dendritic cells and RAW 264.7 macrophages were cultured for 72 h under the indicated conditions (Only Media, SGE, ZIKV, or ZIKV+SGE at different concentrations). Cytokine concentrations [tumour necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), IL-10, IL-12] were quantified in culture supernatants by enzyme-linked immunosorbent assay (ELISA). In JAWS II cells (A-D), significant differences were observed for TNF-α, IL-1β, and IL-12, while IL-10 levels remained stable. In RAW 264.7 cells (E-H), SGE enhanced TNF-α, IL-1β, and IL-10 secretion, whereas IL-12 showed no significant modulation. Data represent individual values from independent experiments with bars indicating mean ± standard error of the mean (SEM). Statistical analysis was performed using Kruskal-Wallis with Dunn’s post-hoc test; p < 0.05 was considered significant.

Article Snippet: Quantification of ZIKV RNA was performed using the AllplexTM Zika Virus Assay (Seegene®), a multiplex Real-Time PCR kit designed for specific detection of ZIKV RNA.

Techniques: Virus, Infection, Cell Culture, Enzyme-linked Immunosorbent Assay