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《Vaccine》2021,39(23):3152-3160
PurposePseudomonas aeruginosa (P. aeruginosa) infection is one of the major causes of keratitis. However, effective prophylactic and therapeutic vaccines against P. aeruginosa keratitis have yet to be developed. In this study, we explored the use of P. aeruginosa membrane vesicles (MVs) as a prophylactic vaccine as well as the use of immune sera derived from P. aeruginosa MV-immunized animals as a treatment for P. aeruginosa corneal infections in C57BL/6 mice.MethodsC57BL/6 mice were intramuscularly immunized with P. aeruginosa MVs; the mouse corneas were then scarified and topically infected with several P. aeruginosa strains, followed by determination of corneal clinical score and corneal bacterial load. Next, immune sera derived from P. aeruginosa MV-immunized ICR mice were administered intraperitoneally to naïve C57BL/6 mice, followed by topical P. aeruginosa challenge. Finally, the immune sera were also used as a topical treatment in the mice with established P. aeruginosa corneal infections.ResultsP. aeruginosa-specific IgG and IgA antibodies induced by intramuscular immunization were detected not only in the sera but also in the eye-wash solution. Both active and passive immunization significantly inhibited P. aeruginosa corneal infection. Finally, topical treatment with immune sera in the mice with established P. aeruginosa corneal infections notably decreased the corneal clinical score and corneal bacterial load.ConclusionsP. aeruginosa keratitis can be attenuated by vaccination of P. aeruginosa MVs and topical application of P. aeruginosa MV-specific immune sera.  相似文献   
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Extracellular vesicles (EVs), including microvesicles and exosomes, are small phospholipid vesicles (≤1 μm in diameter) that are present in blood products, accumulate during storage, and have a potential transfusion-related immunomodulatory role. Knowledge of EVs in stored blood products is limited due to the challenges and difficulties in detecting these heterogeneous submicron-sized vesicles. The aim of this study was to assess the impact of different approaches to characterize EVs in stored RBC products. Quantification and size-profiling of EVs in leukoreduced red cell concentrates (RCCs) were examined on day 3, 7, 21, and 42 of storage using tunable resistive plus sensing (TRPS), flow cytometer (FC), and dynamic light scatting (DLS) methods. Using the TRPS method, the concentration of EVs < 200 nm significantly increased throughout storage (p < 0.05). This change in exosome concentration was not detectable with FC or DLS due to limitations in their ability to resolve particles <200 nm and/or accurately determine EV concentration. Both the TRPS and FC demonstrate that the concentration of EVs  200 nm significantly increases in RCCs by day 42/43 compared to EVs present on day 3 (p < 0.001). As the DLS measures the average size of particles in suspension, only an increase in the zeta-average size was observed during storage. EV size and concentration in RBC products is significantly influenced by the length of storage. Overall, this study shows that combining technologies may be important to improve the characterization and study of EVs in stored RCCs.  相似文献   
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Background: Immunotherapy is one promising therapeutic strategy against glioma, an aggressive form of braincancer. Previous studies have demonstrated that multiple tumor antigens exist and can be used to induce tumor specificT cell responses. Furthermore, recently it was shown that TLR4-primed mesenchymal stem cells (MSCs), also knownas MSC1, mostly elaborate pro-inflammatory mediators. Compared to MSCs, MSC-derived microvesicles (MVs) haveadvantageous properties that present them as stable, long lasting effectors with no risk of immune rejection. Therefore,peripheral blood monocyte derived dendritic cells (MoDCs) have been used to load tumor antigens and stimulate Tcell mediated responses in the presence of MSC1-derived MVs in vitro. Methods: The B92 tumor cell line was heatedto 43°C for 90 min prior to preparation of tumor cell lysates. MVs were purified by differential ultracentrifugationafter isolation, stimulation of proliferation and treatment of MSCs. Autologous T cells isolated from non-adherentcells were harvested during the procedure to generate MoDCs and then incubated with heat stressed tumor cell lysatepulsed DCs in the presence of MSC1-derived MVs. T cells were then co-cultured with tumor cells in 96-well plates ata final volume of 200 μl CM at an effector: target ratio of 100:1 to determine their specific cytotoxic activity. Results:Flow cytometric analysis, T cell mediated cytotoxicity showed that heat stressed tumor antigen pulsed MoDCs andMSC1-derived MVs primed T cells elicited non-significantly enhanced cytotoxic activity toward B92 tumor cells(P≥0.05). Conclusion: These findings may offer new insights into tumor antigen presenting technology involvingdendritic cells and MSC1-derived MVs. Further exploration of the potential of such nanoscale particles in immunotherapyand in novel cancer vaccine settings appears warranted.  相似文献   
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近年来, 随着对微囊泡研究的深入, 如何有效获取足量、高纯度的微囊泡成为该领域研究的挑战。微囊泡又称微粒, 是细胞激活、损伤或凋亡后从细胞膜脱落的小囊泡, 直径为100 ~1000 nm。微囊泡可通过不同机制与细胞相互作用, 发挥与来源细胞相似的生物学功能。与此同时, 微囊泡又因其含有天然的细胞膜成分, 具有良好的生物传递性, 成为仿生药物载体研究的新宠。从原理、应用和关键技术等几个方面综述该领域的研究进展, 将目前已有的微囊泡提取及鉴定关键技术做进一步论述, 并对微囊泡应用前景进行展望。  相似文献   
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Background

Cerebral atherosclerosis is the most important mechanism for ischemic stroke. However, specific plasma biomarkers to assess atherosclerosis susceptibility are still lacking. Circulating miRNAs have been shown to be promising biomarkers for various pathologic conditions. We investigated whether plasma miR-126 and miR-143 could be used as biomarkers for identifying and evaluating cerebral atherosclerosis. Results showed that miR-143 and miR-126 might participate in the process of atherosclerosis and were minimally affected by cerebral infarction. Using Pearson correlation analysis, we showed that miR-126 and miR-143 were correlated with the presence and severity of cerebral atherosclerosis. The ability of miR-126 and miR-143 to differentiate atherosclerosis patients from healthy controls was demonstrated via a receiving operating characteristic curve with high specificity and sensitivity. Our data thus indicate that miR-126 and miR-143 may be potential specific biomarkers for atherosclerosis.  相似文献   
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