Tumor-Derived Microvesicles Induce, Expand and Up-Regulate Biological Activities of Human Regulatory T Cells (Treg)

Background: Tumor-derived microvesicles (TMV) or exosomes are present in body fluids of patients with cancer and might be involved in tumor progression. The frequency and suppressor functions of peripheral blood CD4 + CD25 high FOXP3 + Treg are higher in patients with cancer than normal controls. The hypothesis is tested that TMV contribute to induction/ expansion/and activation of human Treg. Methodology/Principal Findings: TMV isolated from supernatants of tumor cells but not normal cells induced the generation and enhanced expansion of human Treg. TMV also mediated conversion of CD4 + CD25 neg T cells into CD4 + CD25 high FOXP3 + Treg. Upon co-incubation with TMV, Treg showed an increased FasL, IL-10, TGF-b1, CTLA-4, granzyme B and perforin expression (p,0.05) and mediated stronger suppression of responder cell (RC) proliferation (p,0.01). Purified Treg were resistant to TMV-mediated apoptosis relative to other T cells. TMV also increased phospho-SMAD2/3 and phospho-STAT3 expression in Treg. Neutralizing Abs specific for TGF-b1 and/or IL-10 significantly inhibited TMV ability to expand Treg. Conclusions/Significance: This study suggests that TMV have immunoregulatory properties. They induce Treg, promote Treg expansion, up-regulate Treg suppressor function and enhance Treg resistance to apoptosis. Interactions of TMV wit

Interaction and uptake of exosomes by ovarian cancer cells

<p>Abstract</p> <p>Background</p> <p>Exosomes consist of membrane vesicles that are secreted by several cell types, including tumors and have been found in biological fluids. Exosomes interact with other cells and may serve as vehicles for the transfer of protein and RNA among cells.</p> <p>Methods</p> <p>SKOV3 exosomes were labelled with carboxyfluoresceine diacetate succinimidyl-ester and collected by ultracentrifugation. Uptake of these vesicles, under different conditions, by the same cells from where they originated was monitored by immunofluorescence microscopy and flow cytometry analysis. Lectin analysis was performed to investigate the glycosylation properties of proteins from exosomes and cellular extracts.</p> <p>Results</p> <p>In this work, the ovarian carcinoma SKOV3 cell line has been shown to internalize exosomes from the same cells via several endocytic pathways that were strongly inhibited at 4°C, indicating their energy dependence. Partial colocalization with the endosome marker EEA1 and inhibition by chlorpromazine suggested the involvement of clathrin-dependent endocytosis. Furthermore, uptake inhibition in the presence of 5-ethyl-N-isopropyl amiloride, cytochalasin D and methyl-beta-cyclodextrin suggested the involvement of additional endocytic pathways. The uptake required proteins from the exosomes and from the cells since it was inhibited after proteinase K treatments. The exosomes were found to be enriched in specific mannose- and sialic acid-containing glycoproteins. Sialic acid removal caused a small but non-significant increase in uptake. Furthermore, the monosaccharides D-galactose, α-L-fucose, α-D-mannose, D-N-acetylglucosamine and the disaccharide β-lactose reduced exosomes uptake to a comparable extent as the control D-glucose.</p> <p>Conclusions</p> <p>In conclusion, exosomes are internalized by ovarian tumor cells via various endocytic pathways and proteins from exosomes and cells are required for uptake. On the other hand, exosomes are enriched in specific glycoproteins that may constitute exosome markers. This work contributes to the knowledge about the properties and dynamics of exosomes in cancer.</p

Membrane vesicles, current state-of-the-art: emerging role of extracellular vesicles

Release of membrane vesicles, a process conserved in both prokaryotes and eukaryotes, represents an evolutionary link, and suggests essential functions of a dynamic extracellular vesicular compartment (including exosomes, microparticles or microvesicles and apoptotic bodies). Compelling evidence supports the significance of this compartment in a broad range of physiological and pathological processes. However, classification of membrane vesicles, protocols of their isolation and detection, molecular details of vesicular release, clearance and biological functions are still under intense investigation. Here, we give a comprehensive overview of extracellular vesicles. After discussing the technical pitfalls and potential artifacts of the rapidly emerging field, we compare results from meta-analyses of published proteomic studies on membrane vesicles. We also summarize clinical implications of membrane vesicles. Lessons from this compartment challenge current paradigms concerning the mechanisms of intercellular communication and immune regulation. Furthermore, its clinical implementation may open new perspectives in translational medicine both in diagnostics and therapy

Cmos rf transmitters with on-chip antenna for passive RFID and iot nodes

The performances of two RF transmitters, monolithically integrated with their antennas on a single CMOS microchip fabricated in a standard 0.35 µm process, are presented. The usage of these architectures in the Internet of Things (IoT) paradigm is envisioned, as part of a custom conceived data transmission system. The implemented circuits use two different directly on–off keying (OOK) modulated oscillator topologies whose outputs are employed to feed two loop antennas. The powering of both transmitters is duty-cycled for reducing the average power consumption to a few tenths of a microwatt, allowing the usage as low-power transmitters for IoT nodes. The integrated loop antennas radiate sufficient power for a few meters’ communication range. The OOK transmitted signal can be easily detected using a commercial receiver

Reconfigurable UHF RFID tag with sensing capabilities

The pervasiveness of RFID in application fields commonly prerogative of other technologies is related to the addition of sensor and computational capabilities to the systems. This paper presents a smart RFID tag based on a custom designed microchip, able to transmit wirelessly the information coming from a series of integrated sensors and complying with part of the RFID UHF EPC-Gen2 standard, developed implementing the state machine on a Field Programmable Gate Array (FPGA) board. The solution provided aims to demonstrate the feasibility to create augmented RFID tags useful in several applications such as a safer and more manageable food supply chain of perishable comestibles, or biomedical sensor data transmitters. The solution conceived, thanks to the possible miniaturization of the FPGA IC, is a practical solution to obtain compact, small size and low cost devices for short range applications. The FPGA-based implementation ensures a low-cost alternative to dedicated ASIC design for achieving high-end features such as bi-directional communication, anti-collision and on-board computational power

An Indoor Ultrasonic System for Autonomous 3-D Positioning

Indoor positioning is an emerging technology with wide applications. Augmented and mixed reality need accurate and real-time positioning of user's limbs or direction of sight in real time, and reference points within the real environment. Positioning is also required for mall navigation, elderly people movements monitoring, and many others contexts. Indoor positioning of mobile units (MUs) can be provided by multilateration techniques that compute the location of an MU starting from distance measurements between the MU and a set of beacons. In this paper, a nonlinear closed-form solution for the trilateration problem is employed to avoid heavy numerical iterative algorithms. The closed-form solution is allowed by a particular arrangement of the beacons, placed at the vertex of a square. A positioning system is presented where MUs, exploiting the light algorithm of the closed-form positioning, are able to autonomously and privately calculate their own positions. A prototype of the positioning system has been designed, realized, and characterized for an average 4× 4 ×3m3 home or office room. It includes four beacons that emit a sequence of ultrasonic chirp signals, several MUs, and a master unit that provides time synchronization via ANT transceivers onboard each MU and the master unit. Thanks to the adopted closed form solution, each tiny battery-operated MU is able to carry out all the computations onboard, including analog and digital signal processing, beacon-MU distance estimation, and finally MU positioning at a rate of 2 Hz