Extracellular vesicles from oral squamous carcinoma cells display pro- and antiangiogenic properties

Abstract Background: A new intercellular communication mode established by neoplastic cells and tumor microenvironment components is based on extracellular vesicles (EVs). However, the biological effects of the EVs released by tumor cells on angiogenesis are not completed understood. Here we aimed to understand the biological effects of EVs isolated from two cell lines of oral squamous cell carcinoma (OSCC) (SCC15 and HSC3) on endothelial cell tubulogenesis. Methods: OSCC-derived EVs were isolated with a polymer-based precipitation method, quantified using nanoparticle tracking analysis and verified for EV markers by dot-blot. Functional assays were performed to assess the angiogenic potential of the OSCC-derived EVs. Results: The results showed that EVs derived from both cell lines displayed typical spherical-shaped morphology and expressed the EV markers CD63 and Annexin II. Although the average particle concentration and size were quite similar, SCC15-derived EVs promoted a pronounced tubular formation associated with significant migration and apoptosis rates of the endothelial cells, whereas EVs derived from HSC3 cells inhibited significantly endothelial cell tubulogenesis and proliferation. Conclusions: The findings of this study reveal that EVs derived from different OSCC cell lines by a polymer-based precipitation method promote pro- or antiangiogenic effects

Influence of previous Zika virus infection on acute dengue episode.

BackgroundThe co-circulation of flaviviruses in tropical regions has led to the hypothesis that immunity generated by a previous dengue infection could promote severe disease outcomes in subsequent infections by heterologous serotypes. This study investigated the influence of antibodies generated by previous Zika infection on the clinical outcomes of dengue infection.Methodology/principal findingsWe enrolled 1,043 laboratory confirmed dengue patients and investigated their prior infection to Zika or dengue. Severe forms of dengue disease were more frequent in patients with previous Zika infection, but not in those previously exposed to dengue.Conclusions/significanceOur findings suggest that previous Zika infection may represent a risk factor for subsequent severe dengue disease, but we did not find evidence of antibody-dependent enhancement (higher viral titer or pro-inflammatory cytokine overexpression) contributing to exacerbation of the subsequent dengue infection

Screening of reactive peptides using monoclonal antibodies.

(A) Heatmap matrix of peptides assessed with monoclonal specific antibodies against ZIKV or DENV in an indirect IgG ELISA platform. Highest scores were selected for soluble synthesis. Peptide ZV-53 was not stable in soluble form and excluded from analysis. (B) IgG pepELISA for validation of soluble peptides. Serum samples from patients validated with PRNT were evaluated on plates with DV-15, DV-20, ZV-54, and ZV-107.</p

Codebook.

BackgroundThe co-circulation of flaviviruses in tropical regions has led to the hypothesis that immunity generated by a previous dengue infection could promote severe disease outcomes in subsequent infections by heterologous serotypes. This study investigated the influence of antibodies generated by previous Zika infection on the clinical outcomes of dengue infection.Methodology/Principal findingsWe enrolled 1,043 laboratory confirmed dengue patients and investigated their prior infection to Zika or dengue. Severe forms of dengue disease were more frequent in patients with previous Zika infection, but not in those previously exposed to dengue.Conclusions/SignificanceOur findings suggest that previous Zika infection may represent a risk factor for subsequent severe dengue disease, but we did not find evidence of antibody-dependent enhancement (higher viral titer or pro-inflammatory cytokine overexpression) contributing to exacerbation of the subsequent dengue infection.</div

Distribution of dengue-confirmed cases in São José do Rio Preto, São Paulo State, during 2019 epidemic, according to months and epidemiological weeks.

Distribution of dengue-confirmed cases in São José do Rio Preto, São Paulo State, during 2019 epidemic, according to months and epidemiological weeks.</p

Differential diagnostic performance of the pepELISA and standard NS1 ELISA using validated samples.

Receiver Operating Characteristic (ROC) curves of pepELISA for the peptides (A) DV-15; (B) DV-20; (C) ZV-54; and standard ELISA against (D) DENV NS1 mix, and (E) ZIKV NS1. ‘Control’ identifies samples considered true negatives, and ‘Patients’ identifies samples considered true positives. Performance is demonstrated in True Positive Rate (Sensitivity %) versus False Positive Rate (100%—Specificity %). (TIF)</p

Characteristics of 1,043 dengue-confirmed cases eligible between December 2018 and November 2019, in São José do Rio Preto, São Paulo State.

Characteristics of 1,043 dengue-confirmed cases eligible between December 2018 and November 2019, in São José do Rio Preto, São Paulo State.</p

Measure of differential diagnostic performance for DENV NS1 protein, ZIKV NS1 protein, and for peptides DV-15, DV-20, and ZV-54.

Measure of differential diagnostic performance for DENV NS1 protein, ZIKV NS1 protein, and for peptides DV-15, DV-20, and ZV-54.</p

Flow of sample selection for dengue-suspected cases with up to 7 days of symptoms onset.

Flow of sample selection for dengue-suspected cases with up to 7 days of symptoms onset.</p

Dataset.

Deidentified data from samples included in the study. (XLSX)</p