Viral Infection Results in Massive CD8+ T Cell Expansion and Mortality in Vaccinated Perforin-Deficient Mice

AbstractPerforin-mediated cytotoxicity is essential for clearance of primary LCMV infection. BALB/c-perforin-deficient (PKO) mice survived LCMV infection by deleting NP118-specific CD8+ T cells whereas vaccination of PKO mice with Listeria expressing NP118 generated a stable memory CD8+ T cell population. However, >85% of vaccinated BALB/c-PKO mice died after LCMV infection. Mortality was associated with enormous expansion of NP118-specific CD8+ T cells in both lymphoid and nonlymphoid tissues and aberrant CD8+ T cell cytokine production. Depletion of CD8+ T cells or treatment with anti-IFNγ antibody rescued vaccinated mice from mortality. Thus, perforin was essential for resistance to secondary LCMV infection, and, in the absence of perforin, vaccination resulted in lethal disease mediated by dysregulated CD8+ T cell expansion and cytokine production

Design, assembly, and validation of a nose-only inhalation exposure system for studies of aerosolized viable influenza H5N1 virus in ferrets

<p>Abstract</p> <p>Background</p> <p>The routes by which humans acquire influenza H5N1 infections have not been fully elucidated. Based on the known biology of influenza viruses, four modes of transmission are most likely in humans: aerosol transmission, ingestion of undercooked contaminated infected poultry, transmission by large droplets and self-inoculation of the nasal mucosa by contaminated hands. In preparation of a study to resolve whether H5N1 viruses are transmissible by aerosol in an animal model that is a surrogate for humans, an inhalation exposure system for studies of aerosolized H5N1 viruses in ferrets was designed, assembled, and validated. Particular attention was paid towards system safety, efficacy of dissemination, the viability of aerosolized virus, and sampling methodology.</p> <p>Results</p> <p>An aerosol generation and delivery system, referred to as a Nose-Only Bioaerosol Exposure System (NBIES), was assembled and function tested. The NBIES passed all safety tests, met expected engineering parameters, required relatively small quantities of material to obtain the desired aerosol concentrations of influenza virus, and delivered doses with high-efficacy. Ferrets withstood a mock exposure trial without signs of stress.</p> <p>Conclusions</p> <p>The NBIES delivers doses of aerosolized influenza viruses with high efficacy, and uses less starting material than other similar designs. Influenza H5N1 and H3N2 viruses remain stable under the conditions used for aerosol generation and sample collection. The NBIES is qualified for studies of aerosolized H5N1 virus.</p

Feeding dynamics of Northwest Atlantic small pelagic fishes

Author Posting. © The Author(s), 2018. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in Progress in Oceanography 165 (2018): 52-62, doi:10.1016/j.pocean.2018.04.014.Small pelagic fishes represent a critical link between zooplankton and large predators. Yet, the taxonomic resolution of the diets of these important fishes is often limited, especially in the Northwest Atlantic. We examined the diets, along with stable isotope signatures, of five dominant small pelagic species of the Northeast US continental shelf ecosystem (Atlantic mackerel Scomber scombrus, Atlantic herring Clupea harengus, alewife Alosa pseudoharengus, blueback herring Alosa aestivalis, and Atlantic butterfish Peprilus triacanthus). Diet analyses revealed strong seasonal differences in most species. Small pelagic fishes predominantly consumed Calanus copepods, small copepod genera (Pseudocalanus/Paracalanus/Clausocalanus), and Centropages copepods in the spring, with appendicularians also important by number for most species. Krill, primarily Meganyctiphanes norvegica, and hyperiid amphipods of the genera Hyperia and Parathemisto were common in the stomach contents of four of the five species in the fall, with hyperiids common in the stomach contents of butterfish in both seasons and krill common in the stomach contents of alewife in both seasons. Depth and region were also found to be sources of variability in the diets of Atlantic mackerel, Atlantic herring, and alewife (region but not depth) with krill being more often in the diet of alewife in more northerly locations, primarily the Gulf of Maine. Stable isotope data corroborate the seasonal differences in diet but overlap of isotopic niche space contrasts that of dietary overlap, highlighting the differences in the two methods. Overall, the seasonal variability and consumer-specific diets of small pelagic fishes are important for understanding how changes in the zooplankton community could influence higher trophic levels.Funding for this work was primarily through a US National Science Foundation (NSF) OCE-RIG grant (OCE 1325451) to JKL, with additional support from NOAA through the Cooperative Institute for the North Atlantic Region (CINAR) under Cooperative Agreement NA14OAR4320158 in the form a CINAR Fellow Award (JKL), an NSF Long-term Ecological Research grant for the Northeast US Shelf Ecosystem (OCE 1655686; JKL), a Hendrix College summer research award (ZRK), and an NSF REU-supported Woods Hole Oceanographic Institution Summer Student Fellowship (SLH)

IL-4 sensitivity shapes the peripheral CD8\u3csup\u3e+\u3c/sup\u3e T cell pool and response to infection

Previous studies have revealed that a population of innate memory CD8+ T cells is generated in response to IL-4, first appearing in the thymus and bearing high expression levels of Eomesodermin (Eomes) but not T-bet. However, the antigen specificity and functional properties of these cells is poorly defined. In this study, we show that IL-4 regulates not only the frequency and function of innate memory CD8+ T cells, but also regulates Eomes expression levels and functional reactivity of naive CD8+ T cells. Lack of IL-4 responsiveness attenuates the capacity of CD8+ T cells to mount a robust response to lymphocytic choriomeningitis virus infection, with both quantitative and qualitative effects on effector and memory antigen-specific CD8+ T cells. Unexpectedly, we found that, although numerically rare, memory phenotype CD8+ T cells in IL-4Rα–deficient mice exhibited enhanced reactivity after in vitro and in vivo stimulation. Importantly, our data revealed that these effects of IL-4 exposure occur before, not during, infection. Together, these data show that IL-4 influences the entire peripheral CD8+ T cell pool, influencing expression of T-box transcription factors, functional reactivity, and the capacity to respond to infection. These findings indicate that IL-4, a canonical Th2 cell cytokine, can sometimes promote rather than impair Th1 cell–type immune responses

Defining the role of CD69 in the formation of resident memory CD8+ T cells

Resident memory CD8+ T cells (TRM) reside in nonlymphoid tissues. There, they play a key role in preventing reinfection by exerting cytotoxic and inflammatory functions upon exposure to previously encountered pathogens. CD69 is often used as a definitive marker of TRM cells. CD69’s interaction with the G-protein-coupled receptor S1PR1 has been identified as one mechanism by which CD69 can regulate tissue residency. However, the functional requirement for CD69 in promoting the generation and maintenance of CD8+ TRM under a wide variety of circumstances remains unclear. We explored the role of CD69 in tissue residency using co-transfer of antigen specific CD69 sufficient and deficient CD8+ T cells in the context of acute LCMV, Influenza, and VSV infections. Strikingly, we found that CD69 was not necessary for TRM establishment in most tissues, although it can promote TRM localization under some circumstances. This seems to be influenced by the focal point of infection. Interestingly, the kidney appears to rely on CD69 for tissue residency with every model pathogen examined. We propose that the requirement for CD69 is context dependent rather than absolute, and that a combination of factors, including tissue microenvironment and infectious agent, dictate CD69’s influence on development of CD8+ resident memory

Gas-permeable ethylene bags for the small scale cultivation of highly pathogenic avian influenza H5N1 and other viruses in embryonated chicken eggs

<p>Abstract</p> <p>Background</p> <p>Embryonated chicken eggs (ECE) are sometimes used for the primary isolation or passage of influenza viruses, other viruses, and certain bacteria. For small-scale experiments with pathogens that must be studied in biosafety level three (BSL3) facilities, inoculated ECE are sometimes manipulated and maintained in small egg incubators within a biosafety cabinet (BSC). To simplify the clean up and decontamination of an egg incubator in case of egg breakage, we explored whether ethylene breather bags could be used to encase ECE inoculated with pathogens. This concept was tested by determining embryo survival and examining virus yields in bagged ECE.</p> <p>Results</p> <p>Virus yields acceptable for many applications were attained when influenza-, alpha-, flavi-, canine distemper-, and mousepox viruses were propagated in ECE sealed within ethylene breather bags.</p> <p>Conclusions</p> <p>For many small-scale applications, ethylene breather bags can be used to encase ECE inoculated with various viruses.</p

Medication administration errors for older people in long-term residential care

Background Older people in long-term residential care are at increased risk of medication errors. The purpose of this study was to evaluate a computerised barcode medication management system designed to improve drug administrations in residential and nursing homes, including comparison of error rates and staff awareness in both settings. Methods All medication administrations were recorded prospectively for 345 older residents in thirteen care homes during a 3-month period using the computerised system. Staff were surveyed to identify their awareness of administration errors prior to system introduction. Overall, 188,249 attempts to administer medication were analysed to determine the prevalence of potential medication administration errors (MAEs). Error classifications included attempts to administer medication at the wrong time, to the wrong person or discontinued medication. Analysis compared data at residential and nursing home level and care and nursing staff groups. Results Typically each resident was exposed to 206 medication administration episodes every month and received nine different drugs. Administration episodes were more numerous (p < 0.01) in nursing homes (226.7 per resident) than in residential homes (198.7). Prior to technology introduction, only 12% of staff administering drugs reported they were aware of administration errors being averted in their care home. Following technology introduction, 2,289 potential MAEs were recorded over three months. The most common MAE was attempting to give medication at the wrong time. On average each resident was exposed to 6.6 potential errors. In total, 90% of residents were exposed to at least one MAE with over half (52%) exposed to serious errors such as attempts to give medication to the wrong resident. MAEs rates were significantly lower (p < 0.01) in residential homes than nursing homes. The level of non-compliance with system alerts was low in both settings (0.075% of administrations) demonstrating virtually complete error avoidance. Conclusion Potentially inappropriate administration of medication is a serious problem in long-term residential care. A computerised barcode system can accurately and automatically detect inappropriate attempts to administer drugs to residents. This tool can reliably be used by care staff as well as nurses to improve quality of care and patient safety