Es present on EVs between cell lines. Summary/Conclusion: EVQuant is really a rapid, robust, extensively accessible assay with all the following benefits; the ability to detect vesicles down to 50 nm in size, no EV isolation/purification needed, and also the possibility to execute multicolor imaging. The capability to detect EV sub-populations according to particular biomarkers plus the possibility to analyse EV samples in high-throughput, makes EVQuant a suitable candidate for implementation inside a clinical setting. Funding: This project is funded by Prostate Cancer UK (G2012-36)OS26.Extracellular vesicle and miRNA profiling in the primate cervicovaginal compartment reveal possible anti-HIV defences Zezhou Zhao, Dillon Muth, Kathleen Mulka, Bonita Powell, Kelly Metcalf Pate, Zhaohao Liao and Kenneth Witwer The Johns Hopkins University College of Medicine, MD, USALBO.Higher sensitivity, quantitative epitope evaluation of plasma EVs by flow cytometry Aizea Morales-Kastresana1, Xiaomei Yan2, Shaobin Zhu3, Katherine McKinnon4, William Telford5, Veena Kapoor5, Jay A. Berzofsky4 and Jennifer C. JonesNational Cancer Institute, National Institutes of Overall health; 2Xiamen University; Orthopoxvirus site nanoFCM, Inc., Fujian, Chyina; 4National Cancer Institute, Vaccine ADC Linker Chemical Biological Activity Branch; five National Institutes of HealthIntroduction: We previously observed alterations in all round concentration of extracellular particles such as extracellular vesicles (EVs) recovered from cervicovaginal lavage (CVL) and also other fluids in endometriosis and in HIV/SIV infection. Right here, we further characterise EVs released through the menstrual cycle and retroviral infection and create a reference profile of smaller RNAs from the primate cervicovaginal compartment. Solutions: CVL of rhesus macaques, previously collected over the course of five weeks, were subjected to differential centrifugation to enrich EVs. Characterisation was performed by NTA and WB for EV markers. A medium-throughput qPCR platform was applied to profile miRNAs in CVL, swabbed secretions, enriched EVs and biopsy samples, and results were validated by person qPCR. miRNA function in relation to HIV infection was assessed in monocyte-derived macrophages using HIV-1 BaL or green fluorescent protein-encoding HIV. Outcomes: EVs with typical EV markers have been effectively enriched from CVL. On the other hand, miRNAs were present predominantly in EV-depleted fractions of CVL, not in EV-enriched centrifuge pellets. Essentially the most abundant miRNAs across fractions have been miRs-223, -203, -24, -150, -146a, -21, -222, -92a, -17 and -106a, with only a couple of miRNAs enriched in EVs. Surprisingly, couple of miRNAs profile changes had been observed through the menstrual cycle or throughout SIV infection, in either CVL or EVs. However, quite a few abundant CVL miRNAs, like miR-223, might be generally protective against retroviral infection, as recommended by in vitro infection assays. Conclusions: We have established a miRNA profile of CVL fractions and probed the overlap of CVL and EV miRNAs with those identified in secretions and epithelial tissues. Though menstrual cycle and SIV infection have only minor effects on CVL miRNA profiles, CVL miRNAs may well contribute to antiviral defences. Additional studies are underway to elucidate the function of EV modest RNAs in defending against retroviral reproductive tract infection.Introduction: Most conventional flow cytometers are unable to resolve person 30 200 nm extracellular vesicles (EVs), which are most likely to carry fewer than 100 copies of any particular epitope. Commonly, these EVs are usually not only.