Immunomodulatory Effect of Propolis on Foxp3 Gene Expression in Human Peripheral Blood Mononuclear Cells Stimulated in vitro with Pseudomonas Aeruginosa Ag.

Immune balance during infection is critical for both supporting the defense of the immune system of the body and preventing an overly aggressive immune response. Foxp3, a transcription factor of regulatory T cells, plays a critical role in balancing the immune system of the body. Propolis has been shown to affect Foxp3 expression. This study aimed to verify the effect of propolis extracts on in vitro Foxp3 gene expression in peripheral blood mononuclear cells (PBMCs) stimulated with Pseudomonas aeruginosa Ag. In this study, a total of 20 apparently healthy volunteers were included, with 10 males and 10 females within the age range of 20-40 years old. Five ml of blood were drawn from each participant to assess Foxp3 gene expression in PBMCs using density gradient lymphoprep and stimulated with P.aeruginosa lipopolysaccharide (LPS) in vitro. The samples were divided into four distinct groups as follows: LPS stimulated PBMCs, ethanol-extracted propolis (EEP) + LPS stimulated PBMCs, and water-extracted propolis (WEP) + LPS stimulated PBMCs and PBMCs as the control group. The Foxp3 gene expression level was estimated in all four groups following a period of 48 h of cultivation by real-time polymerase chain reaction technique using SYBR green dye. Results of the study indicated that propolis had a great effect on the mRNA Foxp3 expression. Both EEP and WEP had immunomodulatory effects through the Foxp3 mRNA expression, both the EEP and WEP could significantly inhibit Foxp3 mRNA gene expression by human PBMCs after stimulation with pseudomonas Ag in vitro. Propolis exhibited an immunoregulatory effect which was the same with ethanol and water extracts on Foxp3 mRNA gene expression.

Amotosalen and ultraviolet A light efficiently inactivate MERS-coronavirus in human platelet concentrates.

OBJECTIVE: This study aimed to assess the efficacy of the INTERCEPT™ Blood System [amotosalen/ultraviolet A (UVA) light] to reduce the risk of Middle East respiratory syndrome-Coronavirus (MERS-CoV) transmission by human platelet concentrates. BACKGROUND: Since 2012, more than 2425 MERS-CoV human cases have been reported in 27 countries. The infection causes acute respiratory disease, which was responsible for 838 deaths in these countries, mainly in Saudi Arabia. Viral genomic RNA was detected in whole blood, serum and plasma of infected patients, raising concerns of the safety of blood supplies, especially in endemic areas. METHODS: Four apheresis platelet units in 100% plasma were inoculated with a clinical MERS-CoV isolate. Spiked units were then treated with amotosalen/UVA to inactivate MERS-CoV. Infectious and genomic viral titres were quantified by plaque assay and quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR). Inactivated samples were successively passaged thrice on Vero E6 cells to exclude the presence of residual replication-competent viral particles in inactivated platelets. RESULTS: Complete inactivation of MERS-CoV in spiked platelet units was achieved by treatment with Amotosalen/UVA light with a mean log reduction of 4·48 ± 0·3. Passaging of the inactivated samples in Vero E6 showed no viral replication even after nine days of incubation and three passages. Viral genomic RNA titration in inactivated samples showed titres comparable to those in pre-treatment samples. CONCLUSION: Amotosalen and UVA light treatment of MERS-CoV-spiked platelet concentrates efficiently and completely inactivated MERS-CoV infectivity (>4 logs), suggesting that such treatment could minimise the risk of transfusion-related MERS-CoV transmission.