Integration of a virus membrane protein into the lipid bilayer of target cells as a prerequisite for immune cytolysis

Structural requirements for membrane antigens on target cells to mediate immune cytolysis were studied in a model system with purified membrane proteins from Semliki Forest virus (SFV). These SFV spike proteins were isolated in the form of detergent- and lipid-free protein micelles (29S complexes) or, after reconstitution into lipid vesicles, in the form of virosomes. Both the 29S complexes and the virosomes were found to bind well to murine tumor cells (P815 or Eb). When these cells, however, were used as target cells in complement-dependent lysis or in antibody-dependent cell- mediated cytotoxicity assays in the presence of anti-SFV serum, they were not lysed, although they effectively bound the antibody and consumed complement. The same tumor cells infected with SFV served as positive controls in both assays. Different results were obtained when inactivated Sendai virus was added as a fusion reagent to the cells coated with either virosomes or 29S complexes. Under these conditions the virosome-coated cells became susceptible to SFV- specific lysis, whereas the 29S complex-coated cells remained resistant. Evidence that the susceptibility to lysis ofvirosome-coated cells was dependent on active fusion and, therefore, integration of the viral antigens into the lipid bilayer of the target cells was derived from control experiments with enzyme-treated Sendai virus preparations. The 29S complexes and the virosomes partially and selectively blocked the target cell lysis by anti-H-2 sera but not by anti-non-H-2 sera confirming our previous finding that major histocompatibility antigens serve as receptors for SFV. The general significance of these findings for mechanisms of immune cytolysis is dicussed

WebSOS: An Overlay-based System For Protecting Web Servers From Denial of Service Attacks

We present WebSOS, a novel overlay-based architecture that provides guaranteed access to a web server that is targeted by a denial of service (DoS) attack. Our approach exploits two key characteristics of the web environment: its design around a human–centric interface, and the extensibility inherent in many browsers through downloadable "applets." We guarantee access to a web server for a large number of previously unknown users, without requiring pre-existing trust relationships between users and the system, by using reverse Graphic Turing Tests. Furthermore, our system makes it easy for service providers to charge users, providing incentives to a commercial offering of the service. Users can dynamically decide whether to use the WebSOS overlay, based on the prevailing network conditions. Our prototype requires no modifications to either servers or browsers, and makes use of Graphical Turing Tests, web proxies, and client authentication using the SSL/TLS protocol, all readily supported by modern browsers. We then extend this system with a credential-based micropayment scheme that combines access control and payment authorization in one operation. Turing tests ensure that malicious code, such as a worm, cannot abuse a user's micropayment wallet. We use the WebSOS prototype to conduct a performance evaluation over the Internet using PlanetLab, a testbed for experimentation with network overlays. We determine the end-to-end latency using both a chord-based approach and our shortcut extension. Our evaluation shows the latency increase by a factor of 7 and 2 respectively, confirming our simulation results

Comparing the immune response to a novel intranasal nanoparticle PLGA vaccine and a commercial BPI3V vaccine in dairy calves

peer-reviewedBackground There is a need to improve vaccination against respiratory pathogens in calves by stimulation of local immunity at the site of pathogen entry at an early stage in life. Ideally such a vaccine preparation would not be inhibited by the maternally derived antibodies. Additionally, localized immune response at the site of infection is also crucial to control infection at the site of entry of virus. The present study investigated the response to an intranasal bovine parainfluenza 3 virus (BPI3V) antigen preparation encapsulated in PLGA (poly dl-lactic-co-glycolide) nanoparticles in the presence of pre-existing anti-BPI3V antibodies in young calves and comparing it to a commercially available BPI3V respiratory vaccine. Results There was a significant (P < 0.05) increase in BPI3V-specific IgA in the nasal mucus of the BPI3V nanoparticle vaccine group alone. Following administration of the nanoparticle vaccine an early immune response was induced that continued to grow until the end of study and was not observed in the other treatment groups. Virus specific serum IgG response to both the nanoparticle vaccine and commercial live attenuated vaccine showed a significant (P < 0.05) rise over the period of study. However, the cell mediated immune response observed didn’t show any significant rise in any of the treatment groups. Conclusion Calves administered the intranasal nanoparticle vaccine induced significantly greater mucosal IgA responses, compared to the other treatment groups. This suggests an enhanced, sustained mucosal-based immunological response to the BPI3V nanoparticle vaccine in the face of pre-existing antibodies to BPI3V, which are encouraging and potentially useful characteristics of a candidate vaccine. However, ability of nanoparticle vaccine in eliciting cell mediated immune response needs further investigation. More sustained local mucosal immunity induced by nanoparticle vaccine has obvious potential if it translates into enhanced protective immunity in the face of virus outbreak

Interaction of perceptual grouping and crossmodal temporal capture in tactile apparent-motion

Previous studies have shown that in tasks requiring participants to report the direction of apparent motion, task-irrelevant mono-beeps can &quot;capture'' visual motion perception when the beeps occur temporally close to the visual stimuli. However, the contributions of the relative timing of multimodal events and the event structure, modulating uni- and/or crossmodal perceptual grouping, remain unclear. To examine this question and extend the investigation to the tactile modality, the current experiments presented tactile two-tap apparent-motion streams, with an SOA of 400 ms between successive, left-/right-hand middle-finger taps, accompanied by task-irrelevant, non-spatial auditory stimuli. The streams were shown for 90 seconds, and participants' task was to continuously report the perceived (left-or rightward) direction of tactile motion. In Experiment 1, each tactile stimulus was paired with an auditory beep, though odd-numbered taps were paired with an asynchronous beep, with audiotactile SOAs ranging from -75 ms to 75 ms. Perceived direction of tactile motion varied systematically with audiotactile SOA, indicative of a temporal-capture effect. In Experiment 2, two audiotactile SOAs-one short (75 ms), one long (325 ms)-were compared. The long-SOA condition preserved the crossmodal event structure (so the temporal-capture dynamics should have been similar to that in Experiment 1), but both beeps now occurred temporally close to the taps on one side (even-numbered taps). The two SOAs were found to produce opposite modulations of apparent motion, indicative of an influence of crossmodal grouping. In Experiment 3, only odd-numbered, but not even-numbered, taps were paired with auditory beeps. This abolished the temporal-capture effect and, instead, a dominant percept of apparent motion from the audiotactile side to the tactile-only side was observed independently of the SOA variation. These findings suggest that asymmetric crossmodal grouping leads to an attentional modulation of apparent motion, which inhibits crossmodal temporal-capture effects

Integration of a virus membrane protein intothe lipid bilayer of target cells as a prerequisite for immune cytolysis. Specific cytolysis after virosome- target cell fusion

Structural requirements for membrane antigens on target cells to mediate immune cytolysis were studied in a model system with purified membrane proteins from Semliki Forest virus (SFV). These SFV spike proteins were isolated in the form of detergent- and lipid-free protein micelles (29S complexes) or, after reconstitution into lipid vesicles, in the form of virosomes. Both the 29S complexes and the virosomes were found to bind well to murine tumor cells (P815 or Eb). When these cells, however, were used as target cells in complement-dependent lysis or in antibody-dependent cell- mediated cytotoxicity assays in the presence of anti-SFV serum, they were not lysed, although they effectively bound the antibody and consumed complement. The same tumor cells infected with SFV served as positive controls in both assays. Different results were obtained when inactivated Sendai virus was added as a fusion reagent to the cells coated with either virosomes or 29S complexes. Under these conditions the virosome-coated cells became susceptible to SFV- specific lysis, whereas the 29S complex-coated cells remained resistant. Evidence that the susceptibility to lysis ofvirosome-coated cells was dependent on active fusion and, therefore, integration of the viral antigens into the lipid bilayer of the target cells was derived from control experiments with enzyme-treated Sendai virus preparations. The 29S complexes and the virosomes partially and selectively blocked the target cell lysis by anti-H-2 sera but not by anti-non-H-2 sera confirming our previous finding that major histocompatibility antigens serve as receptors for SFV. The general significance of these findings for mechanisms of immune cytolysis is dicussed

Grouping by feature of cross-modal flankers in temporal ventriloquism

Signals in one sensory modality can influence perception of another, for example the bias of visual timing by audition: temporal ventriloquism. Strong accounts of temporal ventriloquism hold that the sensory representation of visual signal timing changes to that of the nearby sound. Alternatively, underlying sensory representations do not change. Rather, perceptual grouping processes based on spatial, temporal, and featural information produce best-estimates of global event properties. In support of this interpretation, when feature-based perceptual grouping conflicts with temporal information-based in scenarios that reveal temporal ventriloquism, the effect is abolished. However, previous demonstrations of this disruption used long-range visual apparent-motion stimuli. We investigated whether similar manipulations of feature grouping could also disrupt the classical temporal ventriloquism demonstration, which occurs over a short temporal range. We estimated the precision of participants’ reports of which of two visual bars occurred first. The bars were accompanied by different cross-modal signals that onset synchronously or asynchronously with each bar. Participants’ performance improved with asynchronous presentation relative to synchronous - temporal ventriloquism - however, unlike the long-range apparent motion paradigm, this was unaffected by different combinations of cross-modal feature, suggesting that featural similarity of cross-modal signals may not modulate cross-modal temporal influences in short time scales

WebSOS: Protecting Web Servers From DDoS Attacks

We present the WebSOS architecture, a mechanism for countering denial of service (DoS) attacks against web servers. WebSOS uses a combination of overlay networking, content-based routing, and aggressive packet filtering to guarantee access to a service that is targeted by a DoS attack. Our approach requires no modifications to servers or browsers, and makes use of the web proxy feature and TLS client authentication supported by modern browsers. We use a WebSOS prototype to conduct a preliminary performance evaluation both on the local area network and over the Internet using PlanetLab, a testbed for experimentation with network overlays. We determine the end-to-end latency imposed by the architecture to increase by a factor of 5 on average. We conclude that this overhead is reasonable in the context of a determined DoS attack

Using Graphic Turing Tests To Counter Automated DDoS Attacks Against Web Servers

We present WebSOS, a novel overlay-based architecture that provides guaranteed access to a web server that is targeted by a denial of service (DoS) attack. Our approach exploits two key characteristics of the web environment: its design around a human-centric interface, and the extensibility inherent in many browsers through downloadable "applets." We guarantee access to a web server for a large number of previously unknown users, without requiring pre-existing trust relationships between users and the system.Our prototype requires no modifications to either servers or browsers, and makes use of graphical Turing tests, web proxies, and client authentication using the SSL/TLS protocol, all readily supported by modern browsers. We use the WebSOS prototype to conduct a performance evaluation over the Internet using PlanetLab, a testbed for experimentation with network overlays. We determine the end-to-end latency using both a Chord-based approach and our shortcut extension. Our evaluation shows the latency increase by a factor of 7 and 2 respectively, confirming our simulation results