Early replication in pulmonary B cells after infection with marek's disease herpesvirus by the respiratory route

Natural infection with Marek's disease virus occurs through the respiratory mucosa after chickens inhale dander shed from infected chickens. The early events in the lung following exposure to the feather and squamous epithelial cell debris containing the viral particles remain unclear. In order to elucidate the virological and immunological consequences of MDV infection for the respiratory tract, chickens were infected by intratracheal administration of infective dander. Differences between susceptible and resistant chickens were immediately apparent, with delayed viral replication and earlier onset of interferon (IFN)-γ production in the latter. CD4+ and CD8 + T cells surrounded infected cells in the lung. Although viral replication was evident in macrophages, pulmonary B cells were the main target cell type in susceptible chickens following intratracheal infection with MDV. In accordance, depletion of B cells curtailed viremia and substantially affected pathogenesis in susceptible chickens. Together the data described here demonstrate the role of pulmonary B cells as the primary and predominant target cells and their importance for MDV pathogenesis. © 2009, Mary Ann Liebert, Inc.

Chicken TREM-B1, an Inhibitory Ig-Like Receptor Expressed on Chicken Thrombocytes

Triggering receptors expressed on myeloid cells (TREM) form a multigene family of immunoregulatory Ig-like receptors and play important roles in the regulation of innate and adaptive immunity. In chickens, three members of the TREM family have been identified on chromosome 26. One of them is TREM-B1 which possesses two V-set Ig-domains, an uncharged transmembrane region and a long cytoplasmic tail with one ITSM and two ITIMs indicating an inhibitory function. We generated specific monoclonal antibodies by immunizing a Balb/c mouse with a TREM-B1-FLAG transfected BWZ.36 cell line and tested the hybridoma supernatants on TREM-B1-FLAG transfected 2D8 cells. We obtained two different antibodies specific for TREM-B1, mab 7E8 (mouse IgG1) and mab 1E9 (mouse IgG2a) which were used for cell surface staining. Single and double staining of different tissues, including whole blood preparations, revealed expression on thrombocytes. Next we investigated the biochemical properties of TREM-B1 by using the specific mab 1E9 for immunoprecipitation of either lysates of surface biotinylated peripheral blood cells or stably transfected 2D8 cells. Staining with streptavidin coupled horse radish peroxidase revealed a glycosylated monomeric protein of about 50 kDa. Furthermore we used the stably transfected 2D8 cell line for analyzing the cytoplasmic tyrosine based signaling motifs. After pervanadate treatment, we detected phosphorylation of the tyrosine residues and subsequent recruitment of the tyrosine specific protein phosphatase SHP-2, indicating an inhibitory potential for TREM-B1. We also showed the inhibitory effect of TREM-B1 in chicken thrombocytes using a CD107 degranulation assay. Crosslinking of TREM-B1 on activated primary thrombocytes resulted in decreased CD107 surface expression of about 50-70%

Chicken CRTAM Binds Nectin-Like 2 Ligand and Is Upregulated on CD8⁺ αβ and γδ T Lymphocytes with Different Kinetics

During a search for immunomodulatory receptors in the chicken genome, we identified a previously cloned chicken sequence as CRTAM homologue by its overall identity and several conserved sequence features. For further characterization, we generated a CRTAM specific mab. No staining was detectable in freshly isolated cell preparations from thymus, bursa, caecal tonsils, spleen, blood and intestine. Activation of splenocytes with recombinant IL-2 increased rapid CRTAM expression within a 2 h period on about 30% of the cells. These CRTAM+ cells were identified as CD8+ γδ T lymphocytes. In contrast, CRTAM expression could not be stimulated on PBL with IL-2, even within a 48 h stimulation period. As a second means of activation, T cell receptor (TCR) crosslinking using an anti-αβ-TCR induced CRTAM on both PBL and splenocytes. While CRTAM expression was again rapidly upregulated on splenocytes within 2 h, it took 48 h to reach maximum levels of CRTAM expression in PBL. Strikingly, albeit the stimulation of splenocytes was performed with anti-αβ-TCR, CRTAM expression after 2 h was mainly restricted to CD8+ γδ T lymphocytes, however, the longer anti-TCR stimulation of peripheral blood lymphocytes (PBL) resulted in CRTAM expression on αβ T lymphocytes. In order to characterize the potential ligand we cloned and expressed chicken Necl-2, a member of the nectin and nectin-like family which is highly homologous to its mammalian counterpart. Three independent assays including a reporter assay, staining with a CRTAM-Ig fusion protein and a cell conjugate assay confirmed the interaction of CRTAM with Necl-2 which could also be blocked by a soluble CRTAM-Ig fusion protein or a CRTAM specific mab. These results suggest that chicken CRTAM represents an early activation antigen on CD8+ T cells which binds to Necl-2 and is upregulated with distinct kinetics on αβ versus γδ T lymphocytes

Structural and regulatory diversity shape HLA-C protein expression levels

Expression of HLA-C varies widely across individuals in an allele-specific manner. This variation in expression can influence efficacy of the immune response, as shown for infectious and autoimmune diseases. MicroRNA binding partially influences differential HLA-C expression, but the additional contributing factors have remained undetermined. Here we use functional and structural analyses to demonstrate that HLA-C expression is modulated not just at the RNA level, but also at the protein level. Specifically, we show that variation in exons 2 and 3, which encode the α1/α2 domains, drives differential expression of HLA-C allomorphs at the cell surface by influencing the structure of the peptide-binding cleft and the diversity of peptides bound by the HLA-C molecules. Together with a phylogenetic analysis, these results highlight the diversity and long-term balancing selection of regulatory factors that modulate HLA-C expression

Role of Position 627 of PB2 and the Multibasic Cleavage Site of the Hemagglutinin in the Virulence of H5N1 Avian Influenza Virus in Chickens and Ducks

Highly pathogenic H5N1 avian influenza viruses have caused major disease outbreaks in domestic and free-living birds with transmission to humans resulting in 59% mortality amongst 564 cases. The mutation of the amino acid at position 627 of the viral polymerase basic-2 protein (PB2) from glutamic acid (E) in avian isolates to lysine (K) in human isolates is frequently found, but it is not known if this change affects the fitness and pathogenicity of the virus in birds. We show here that horizontal transmission of A/Vietnam/1203/2004 H5N1 (VN/1203) virus in chickens and ducks was not affected by the change of K to E at PB2-627. All chickens died between 21 to 48 hours post infection (pi), while 70% of the ducks survived infection. Virus replication was detected in chickens within 12 hours pi and reached peak titers in spleen, lung and brain between 18 to 24 hours for both viruses. Viral antigen in chickens was predominantly in the endothelium, while in ducks it was present in multiple cell types, including neurons, myocardium, skeletal muscle and connective tissues. Virus replicated to a high titer in chicken thrombocytes and caused upregulation of TLR3 and several cell adhesion molecules, which may explain the rapid virus dissemination and location of viral antigen in endothelium. Virus replication in ducks reached peak values between 2 and 4 days pi in spleen, lung and brain tissues and in contrast to infection in chickens, thrombocytes were not involved. In addition, infection of chickens with low pathogenic VN/1203 caused neuropathology, with E at position PB2-627 causing significantly higher infection rates than K, indicating that it enhances virulence in chickens

ES2009-90209 WHAT'S LEFT OVER: PROCESS LOADING IN HIGH PERFORMANCE BUILDINGS ES2009-90209

INTRODUCTION As an increasing number of buildings aspire to significant energy reduction and higher Leadership in Energy and Environmental Design (LEED®) certifications, it is clear that non-regulated process loads are the single greatest remaining opportunity in high performance design. Historically outside the purview of state and national energy design standards, the performance of transformers, elevators, escalators, process systems, and consumer information technology underscore a significant challenge and rising opportunity. This paper examines the impact of nonregulated loads on high performance buildings, ranging from first cost to energy performance and occupant behavior. Drawing on their experiences and research, the authors examine how non-regulated loads can be successfully addressed by team members with diverse skills brought together early in collaborative discussions across their traditional disciplines. By cross-connecting strategies for heating, ventilation, and air-conditioning (HVAC) design, electrical engineering, and information technology, the authors will identify why synergy is often more important than efficiency and why the best solution often integrates function, policy, and practice. In addition to discussing examples of non-regulated load solutions that make better use of their resources and lower cost, indirect impacts and benefits that are less tangible in the short term, but of great value in the long term arise. Improved process load performance often means greater resilience to catastrophic events such as earthquakes, terrorist attacks, and hurricanes and reduced economic risks from fuel volatility, carbon tariffs, and cultural creative losses. As building efficiency exceeds 50%, 60% and greater thresholds, our successful response will depend not on reducing traditional end uses such as lighting and air conditioning, but as well on an aggressive, collaborative and integrated approach to process loads. CONTEXT History of Energy Standards and Relationship to Building Performance To understand the significance of process loads in building energy performance, it is important to provide a concise history of energy standards and how the energy standards have been designed for use. National energy standards were first adopted in the United States in the mid 1970's as a response to the oil price shocks and ensuing national outcry for improved energy efficiency. The most recognized national model energy standard American Society of Heating, Refrigerating and Air-Conditioning Engineers/ Illuminating Engineering Society of North America (ASHRAE/IESNA) Standard 90.1 was first published in 1975(now in its 6 th revision period). As an energy standard, Standard 90.1 was used for years by authorities to establish building's compliance with the accepted minimum standard of expectation. The intended use was as a pass/fail model and the notion of percentage improvement above and beyond the minimum was not addressed. In the late 1990's as the US Green Building Council formed and sought a reference benchmark for the sliding scale award of points in it's LEED rating system, standard 90.1 was chosen. Arguably, it was not a difficult choice as it was only in 1998 that the second model national energy standard was published by the International Code Council (ICC) as the International Energy Conservation Code (IECC). The option for non-residential energy codes in the United States (US). Since their adoption, the US energy codes (standards codified and amended by individual states) have saved a great deal of energy. Although excellent documents, there has in hindsight been a gap in their application; that gap is "non-regulated loads". Standard 90.1 separates energy end uses into two categories, those that are "regulated" and those that are "non-regulated". Only regulated loads are subject to the provisions of the energy standard. Regulated energy includes those building energy impacts primarily related to the building envelope, its lighting systems, and its HVAC systems. Non-regulated loads include those impacts from building systems that use energy primarily for process or industrial purposes. Such energy use as results from elevators, building transformers, data centers, consumer electronics, plant and animal life support, and a great deal of laboratory systems. Whereas regulated energy use is typically considered to be under the control of designers and building owners, nonregulated energy use is typically considered to be a constant unavoidable and uncontrollable load, and hence has in many instances largely been ignored. In 2001, energy performance reporting started to shift with the publication of Addendum E to Standard 90.1-1999. This was followed in 2004 by a formal inclusion of Appendix G -Performance Rating Method -in the Standard 90.1 update of the same year. For the first time, energy reporting on building performance was required to include all energy uses, both regulated and non-regulated. However, even with this advance, an energy code for non-regulated energy uses was not applied. Those reporting building performance were required only to include the non-regulated "process" loads, not improve on them. Process Load Fraction in High Performance Buildings As energy saving strategies have continued to focus on envelope optimization, lighting efficiency and progressive HVAC design, not only do non-regulated loads often become a design after-thought, they effectively become the largest remaining slice of energy consumption in high performance buildings, representing the greatest remaining savings opportunity. The U.S. department of energy estimates that nonregulated loads account for 28% of energy consumption in all commercial U.S. buildings OPPORTUNITIES Technology Introduction Technological progress and growing environmental concerns have led manufacturers to develop new transformer technologies intended to reduce inefficiencies inherent in transformer design. But in the last 30 years, the average efficiency of low-voltage, drytype distribution transformers in use has actually decreased, despite technological improvements in efficient transformer design. This is due in part to the fact that increases in efficiency come with tradeoffs such as increases in first cost. Because equipment specifiers are usually not responsible for building utility costs, transformer selection is largely first-cost driven. However, transformers typically run 24 hours per day, every day of the year, for the entire lifetime of a building; furthermore, transformers exhibit a typical lifetime of over 30 years. Because of this, efficiency increases as low as 0.1% can result in significant reductions in the lifetime energy costs of a building, and can lead to significant long-term cost paybacks far exceeding initial cost premiums. Careful examination of the costs and benefits of transformer technologies is required in order to maximize gains in transformer selection. Recently, the National Electrical Manufacturers Association (NEMA) TP-1 efficiency levels have been set as the legally-mandated minimum efficiency levels for low-voltage, dry-type transformers. Minimum efficiency standards for each transformer type governed by TP-1 are set at an indexed load, measured as a percentage of nameplate load capacity. In the case of low-voltage, dry-type transformers, minimum efficiency is specified at 35% of total load. This is in recognition of the fact that, as most industry experts agree, transformers are typically lightly loaded. But a notable study subsequently performed by the Cadmus Group With baseline levels now set at NEMA TP-1 standards, specifiers now must look to transformers that offer further levels of efficiency in order to qualify for environmental credits and incentives. In Arup's study, transformer technologies compared to this baseline design include higher-grade transformers, fan-assisted transformers, and amorphous-core transformers. Technology Overview Fan Assisted: Fan-assisted transformers are a transformer technology that differs from standard transformers by providing fans that assist in cooling during elevated load conditions, essentially boosting the Copyright © 2009 by ASME 2 Downloaded From: https://proceedings.asmedigitalcollection.asme.org on 06/29/2019 Terms of Use: http://www.asme.org/about-asme/terms-of-use transformer capacity. Due to the low average loading of transformers, the load on a given conventional transformer size can typically be accommodated by a smaller fan-assisted transformer size. Analysis of fanassisted transformers reveals efficiency gains over baseline TP-1 levels at loads falling at or below 35% load levels. However, these transformers exhibit a reduced lifespan due to the non-replacement ability of the fan (the shortest life component in the system), leading to greater long-term cost that is not typically cost-effective (with the exception of very lightly-loaded transformers) Fan-assisted transformers do show benefits due to reduced size, but can also exhibit increased heat output and acoustic noise. Due to the very specific scenarios for ideal usage, fan-assisted transformers should be carefully screened prior to project specification. Amorphous Core: Amorphous-core transformers utilize amorphous alloys as the core material instead of the typical silicon steel core of conventional transformers. At low load levels (in the 10-25% range), amorphous core transformers are among the most efficient transformers available. Despite higher first cost, larger size, and increased noise, the long-term cost savings resulting from efficiency increases makes amorphous-core technology an attractive choice for transformer specification. The most significant barrier to their use is lack of widespread availability in the United States. Nonetheless, availability of amorphouscore transformers should be evaluated on a project-byproject basis for potential incorporation now and in the future. CSL 3+: Higher-grade transformers represent transformers designed to operate at efficiency levels specified by the Department of Energy (DOE) that are beyond those of baseline TP-1 levels. These DOE efficiency levels are noted as Class Standard Levels (CSL). TP-1 efficiencies correspond to CSL 1, while higher-grade transformers are usually CSL 3 or higher. They typically represent overall efficiency improvements over NEMA TP-1 design, although efficiency curves can be selected for specific design criteria. Furthermore, typical transformers in applications involving a high proportion of non-linear loads typical of many modern buildings experience a loss in efficiency beyond rated efficiency levels. Standard higher-grade transformers are K-rated, and perform better when compared to typical transformers, minimizing efficiency losses due to significant amounts of non-linear loading. Higher first cost and larger size are disadvantages, while benefits include long-term cost savings and improved harmonic-handling capability. In the majority of typical uses, higher-grade transformers represent costbeneficial selections. Due to the financial and environmental advantages of this technology, highergrade transformers can typically be recommended for project-wide specification. Benefits Both economic and environmental interests present compelling reasons for maximizing transformer energy efficiency. Improved transformer efficiency can result in significant long-term savings due to reduced energy usage, particularly in areas with high energy prices. Utilization of transformers with higher efficiencies can assist in the successful pursuit of LEED, Performance Criteria goals, Savings by Design, and other mandates and incentives, as these are dependent in part on energy efficiency. Improved transformer efficiency can represent a sizeable portion of the energy performance achievement of the building (on the order of 1-2%). Furthermore, intangible benefits can include prestige/demonstration benefits from adoption of new technologies to reduce energy use. Costs The principal barrier to widespread use of transformers with increased efficiency is increased first cost. For higher-grade transformers, however, cost analysis shows simple paybacks of these increased costs in 2-10 years. Given the average lifetime of transformers of over 30 years, high performance transformers can yield substantial return on investment in most applications. Despite verifiable cost analyses, in many cases the equipment specifier is not the same party that will be responsible for utility payments. In these instances, driving down the first cost to a minimum continues to be a priority. Realizing the long-term costsavings involved in the use of higher-grade transformers requires backing from project Owners throughout the lifetime of the project. Other costs are specific to the drawbacks of the type of transformer technology used. Both amorphous core and higher-grade transformers are physically larger than typical TP-1 transformers; for higher-grade transformers, this is typically on the order of a 6-7% size increase. In most designs this is negligible, but if space is at a premium, the increase may represent a cost due to slightly increased electrical room sizes. This cost may sometimes be offset by a level of HVAC reduction due to reduced heat output. Arup has performed cost analysis designed to highlight simple payback periods for (3) common loading scenarios of low-voltage, dry-type transformers. These scenarios have used Powersmiths brand transformers as examples of higher-grade technology, and compared these to conventional TP-1, 150°C rise units as a baseline. Using these assumptions, Arup developed the following payback periods for laboratory, data center, and office scenarios in a project with electrical cost of 11cents/kWh. . The consolidated IT solution centralizes maintenance and results in the shared investment in and operation of applications, storage, and processing. As a result of the annual 12% growth in consolidated IT energy demand, the Environmental Protection Agency (EPA) estimates that the peak demand from data centers in 2008 was equivalent to approximately 15 power plants. Technology Overview The easy win that has had the most publicity is the shift towards virtualization of services where multiple physical servers are migrated to a single server that appears to the network and users as multiple servers. There are in addition a number of less well publicized options for reducing energy consumption in a consolidated technology space such as a data center. The first of these is to run the data center at higher temperatures. Data centers are often run at temperatures of 70 degrees or lower, as a holdover from the days of less reliable cooling infrastructure, allowing more time to respond following mechanical systems failure. ASHRAE has changed their recommendation so that data centers should now run between 68 degrees and 77 degrees, a view also supported by major server Original Equipment Manufacturers (OEM's). Each rise of 1 degree in temperature saves about 4% in cooling costs. The operating temperatures of servers are commonly above 90 degrees, so the increased temperature should not cause performance or warranty issues. The second option is not to try and cool the exhausted air in the data center. Air ducts can be fixed to the back of racks so that the exhausted air goes directly to heat exchangers or is vented externally. This will mean that energy is either partially recovered via the heat exchange process, or no energy is consumed cooling the exhaust air, just venting it. Particularly in temperate climates external air can be used as make up air as it will be cooler than the air being exhausted. The third option is to provide DC powered hardware. All electronics consumes DC power, so the removal of the AC to DC conversion of power supplies removes inefficiencies to result in a more efficient utilization of the power provided to the servers. Most major OEM's have an option for DC power supplies to their equipment. Benefits Each rise of 1 degree in temperature saves about 4% in cooling costs. The operating temperatures of servers are commonly above 90 degrees, so the increased 1 The cloud environment is simply a shift of applications, storage and processing power to an offsite provider rather than hosting the services within an organization and thus the same energy reduction principles apply. temperature should not cause performance or warranty issues. Costs There is no cost to increasing the set point for cooling in a data center type environment. A change to DC power supply to racks is more expensive than a traditional AC power supply. This centers around the lack of a standardized internationally acceptable connector like an IEC plug and the additional costs and risks associated with high current busbars to supply power, something that the telecom industry has moved away from. Edge IT Introduction In addition to the centralized high energy, high availability components of IT infrastructure, there are energy savings to be made in edge devices such as personal computers, cellular phones and chargers, and task lighting. Technology Overview A significant proportion of IT related power consumption occurs in many distributed devices such as PC's, laptops or IP telephones all consume power. While desktop PC's or laptops have visible power cords they also have the inherent inefficiencies of the AC to DC conversion. IP telephony uses Power over Ethernet (IEEE 802.3af) to power the telephones and there are now low power computing devices, normally with very limited processing power than can be powered by this same method. The Power over Ethernet Plus standard (IEEE P802.3at), currently awaiting ratification will increase the power that can be delivered along an Ethernet cable from 13W to 30W and in some cases up to 60W of power. This is more than enough for a more conventional low power laptop type device. As well as a tool for making more efficient use of server hardware, virtualization can be used to allow low powered devices to be the front end for centralized processing. The Cell phone industry has agreed to move towards a standardized mini-USB connector for all cell phones by 2012. Benefits The benefit of moving towards Power over Ethernet for edge connected devices allows heat generating network switches to be located as part of a consolidated centralized IT infrastructure and means less individual power transformers are required, providing that an excess of switch ports is not provided. This greater standardization will allow more efficient cell phone chargers to be developed as economies of scale result. Costs The cost of a power over Ethernet enabled network switch is about 30% more expensive than a non power Copyright © 2009 by ASME 4 Downloaded From: https://proceedings.asmedigitalcollection.asme.org on 06/29/2019 Terms of Use: http://www.asme.org/about-asme/terms-of-use over Ethernet switch. The cost saving comes from less individual discrete power transformers and receptacles required to support them. Vertical Circulation Introduction When considering vertical circulation systems such as elevators and escalators, the most common topic of concern for manufacturers, operators and passengers, is safety. This is reflected in escalator and elevator codes and standards such as the American Society of Mechanical Engineers (ASME) A17 series, escalator and elevator signage, and the general awareness that passengers have about safe escalator and elevator use. With very little emphasis placed on energy efficiency, these systems therefore represent a tremendous savings potential for most multi-level buildings. This section explores the topics of energy efficiency through advances in technology, and energy savings through optimal operation of these systems. Technology Overview Elevators Elevators typically account for 3 -10% of building energy use [3]. However, it can be argued that the real energy fraction is even higher since elevators also account for the following energy end uses and losses within buildings: • The component of HVAC consumption associated with cooling elevator penthouses • Significant heat loss due to stack effect (especially in cold climates) resulting from poor air-tightness of elevator shafts • Elevator lighting, display and passenger entertainment systems The approach to energy efficient design for elevators has therefore not only targeted the minimization of energy use in actually moving elevator cars, but also minimizing heat gain from components such as gears and brake-discs (which otherwise impose a constant cooling load requirement), optimizing elevator lighting, and improving controls to optimize elevator travel logic and sequences. The KONE Machine Room-Less (MRL) elevator [4] is an example of industry targeting elevator inefficiency and HVAC loads due to heat gains. The elevator eliminates the requirement for an elevator penthouse (and subsequent cooling load) altogether by introducing a low friction, gearless and compact hoisting machine installed directly onto the guide rail. Several manufacturers have also started to incorporate energy recovery features such as regenerative braking whereby electric energy is fed back to a buildings electrical system, rather than being dissipated entirely as heat. This results in an HVAC load reduction as well as an energy recovery component and typically results in a 30% reduction in energy use compared to base case geared traction systems Fuzzy logic-based controls are also being employed more commonly in order to optimize elevator travel and avoid unnecessary car movement [6]. These advanced

DEC-205 expression on migrating dendritic cells in afferent lymph

Previous studies have identified a 210 000-molecular weight molecule expressed at a high level on the surface of dendritic cells (DCs) in afferent lymph of cattle and evident on cells with the morphology of DCs in lymphoid tissues. Expression is either absent from other immune cells or is present at a lower level. The molecular weight and cellular distribution suggested that the molecule, called bovine WC6 antigen (workshop cluster), might be an orthologue of human DEC-205 (CD205). To establish whether this was the case, the open reading frame of bovine DEC-205 was amplified, by polymerase chain reaction, from thymic cDNA (accession no. AY264845). The cDNA sequence of bovine DEC-205 had 86% and 78% nucleic acid identity with human and mouse molecules, respectively. COS-7 cells transfected with a plasmid containing the cattle DEC-205 coding region expressed a molecule that stained with WC6-specific monoclonal antibody, showing that ruminant WC6 is an orthologue of DEC-205. Two-colour flow cytometry of mononuclear cells from afferent lymph draining cattle skin, and from blood, confirmed the high level of expression on large cells in lymph that were uniformly DC-LAMP positive and major histocompatibility complex class II positive. Within this DEC-205(+) DC-LAMP(+) population were subpopulations of cells that expressed the mannose receptor or SIRPα. The observations imply that DCs in afferent lymph are all DEC-205(high), but not a uniform population of homogeneous mature DCs