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Pre-existing SARS-CoV-2-specific T cells, but not antibodies, have been detected in some unexposed individuals. This may account for some of the diversity in clinical outcomes ranging from asymptomatic infection to severe COVID-19. Although age is a risk factor for COVID-19, how age affects SARS-CoV-2-specific T cell responses remains unknown. We found that some pre-existing T cell responses to specific SARS-CoV-2 proteins, Spike (S) and Nucleoprotein (N), were significantly lower in elderly donors (>70 years old) who were seronegative for S than in young donors. However, substantial pre-existing T cell responses to the viral membrane (M) protein were detected in some elderly donors. These responses likely compensate for loss of T cell responses specific to S and N. In contrast, young and elderly donors exhibited comparable T cell responses to S, N, and M proteins after infection with SARS-CoV-2. M-specific responses were mediated by CD4 T cells producing interferon-γ in both seronegative and seropositive individuals. T cells in seronegative elderly donors responded to various M-derived peptides, while the response after SARS-CoV-2 infection was apparently focused on a single peptide. These data suggest that diversity of target antigen repertoire for pre-existing SARS-CoV-2-specific T cells declines with age, but the magnitude of pre-existing T cell responses is maintained by T cells reactive to specific viral proteins such as M. A better understanding of the role of pre-existing SARS-CoV-2-specific T cells that are less susceptible to age-related loss may contribute to development of more effective vaccines for elderly people.

During August 2020, we carried out a serological survey among students and employees at the Okinawa Institute of Science and Technology Graduate University (OIST), Japan, testing for the presence of antibodies against SARS-CoV-2, the causative agent of COVID-19. We used a FDA-authorized 2-step ELISA protocol (1, 2) in combination with at-home self-collection of blood samples using a custom low-cost finger prick-based capillary blood collection kit. Although our survey did not find any COVID-19 seropositive individuals among the OIST cohort, it reliably detected all positive control samples obtained from a local hospital and excluded all negatives controls. We found that high serum antibody titers can persist for at least up to 6.5 months post infection. Among our controls, we found strong cross-reactivity of antibodies in samples from a serum pool from two MERS patients in the anti-SARS-CoV-2-S ELISA. Here we show that a centralized ELISA in combination with patient-based capillary blood collection using as little as one drop of blood can reliably assess the seroprevalence among communities. Anonymous sample tracking and an integrated website created a stream-lined procedure. Major parts of the workflow were automated on a liquid handler, demonstrating scalability. We anticipate this concept to serve as a prototype for reliable serological testing among larger populations.

Background Reducing the emissions of greenhouse gases (GHGs) is vital for mitigating climate change and meeting commitments to international agreements such as the COP 21 Paris Agreement of 2015. Agriculture is reported to account for approximately 11 percent of total global GHG emissions such that: the agricultural sector has an important role to play in meeting climate change mitigation objectives. However, there is currently little consensus on how farm management and interventions, along with interactions with in-field variability, such as soil type, affect the production and assimilation of GHGs in arable crop lands. Practical recommendations for farmers are often vague or generalised, and models (e.g. on the amount of nitrogen fertiliser applied) are used despite limited understanding of the influence of local conditions, such as the importance of soil type. Here, we report the findings of a systematic map of the evidence relating to the impact on GHG flux from the in-field management of arable land in temperate regions. Methods We searched for, collated and catalogued research relating to the effects of in-field arable farming practices in temperate systems on GHG emissions. Results from 6 bibliographic databases, a web-based search engine and organisational websites were combined with evidence from stakeholders. Duplicates were removed and the results were then screened for relevance at title and abstract, and full-text levels according to a predefined set of eligibility criteria (following consistency checking). Relevant studies were then coded and their meta-data extracted and used to populate a systematic map database describing each study’s settings, methods and measured outcomes. Results The mapping process identified 538 relevant studies from 351 articles. Nearly all of these (96%) were found from traditional research papers, with 42% from European countries and nearly half (203 studies) lasting for 12 months or less. Over half of all studies (55%) investigated multiple interventions with chemical fertiliser (n = 100), tillage (n = 70), and organic fertiliser (n = 30) the most frequently studied single intervention types. When combining individually studied and multiple interventions, the top three intervention types most frequently studied were: chemical fertiliser (n = 312); organic fertiliser (n = 176) and tillage (n = 158). Nitrous oxide was the most commonly studied outcome, with over double the number of studies compared to carbon dioxide, the next most studied outcome. Sandy loam and silty loam were the most commonly studied soils but there was a good distribution of studies across other types. However, studies predominately focused on humid sub-tropical (Cfa) and temperate oceanic (Cfb) climates, with hot summer Mediterranean (CSa) and warm summer Mediterranean (Csb) climate zones less represented. Conclusions The mapping process identified clusters of research for chemical and organic fertiliser especially in relation to nitrous oxide emissions and for both carbon dioxide and nitrous dioxide in relation to tillage. Therefore, there is potential for further synthesis for these interventions. The spread of research across soil textures and in the humid sub-tropical and temperate oceanic climates may enable further synthesis to provide tailored in-field advice for farmers and provide an evidence base to inform subsidies policy. However, smaller amounts of research relating to biochar, cover crops, crop rotation, and nitrogen inhibitors highlight gaps where further research would be beneficial.

There is mounting evidence that the deoxygenation of coastal marine ecosystems has been underestimated, particularly in the tropics. These physical conditions appear to have far‐reaching consequences for marine communities and have been associated with mass mortalities. Yet little is known about hypoxia in tropical habitats or about the effects it has on reef‐associated benthic organisms. We explored patterns of dissolved oxygen (DO) throughout Almirante Bay, Panama and found a hypoxic gradient, with areas closest to the mainland having the largest diel variation in DO, as well as more frequent persistent hypoxia. We then designed a laboratory experiment replicating the most extreme in situ DO regime found on shallow patch reefs (3 m) to assess the response of the corallivorous fireworm, Hermodice carnaculata to hypoxia. Worms were exposed to hypoxic conditions (8 hr ~ 1 mg/L or 3.2 kPa) 16 times over an 8‐week period, and at 4 and 8 weeks, their oxygen consumption (respiration rates) was measured upon reoxygenation, along with regrowth of severed gills. Exposure to low DO resulted in worms regenerating significantly larger gills compared to worms under normoxia. This response to low DO was coupled with an ability to maintain elevated oxygen consumption/respiration rates after low DO exposure. In contrast, worms from the normoxic treatment had significantly depressed respiration rates after being exposed to low DO (week 8). This indicates that oxygen‐mediated plasticity in both gill morphology and physiology may confer tolerance to increasingly frequent and severe hypoxia in one important coral predator associated with reef decline., There is mounting evidence that the deoxygenation of coastal marine ecosystems has been underestimated, particularly in the tropics. We provide clear evidence that severe diurnal hypoxia on shallow coral reefs is associated with continuous hypoxia trends found in deeper bay water. Additionally, we experimentally show that under hypoxia, the fireworm Hermodice carunculata is able to regenerate significantly larger gills compared to worms under normoxia, as well as rapidly recover metabolically after hypoxia exposure.

We estimated the seroprevalence of anti-SARS-COV-2 IgG in different island groups in Okinawa and described its changes over time. A cross-sectional sero-survey was repeated in three distinct periods between July 2020 and February 2021. A total of 2683 serum samples were collected from six referral medical centers, each covering a separate region in Okinawa. Patients who visited the emergency department for any reason and underwent blood collection were eligible for the study. Samples were analyzed using an FDA-authorized two-step enzyme-linked immunosorbent assay (ELISA) protocol. The case detection ratio was computed by dividing the seroprevalence by the attack rate obtained from publicly available surveillance data. In the main island, the seroprevalence was 0.0% (0/392, 95% CI: 0.0-0.9), 0.6% (8/1448, 0.2-1.1), and 1.4% (8/582, 0.6-2.7) at the 1st, 2nd, and 3rd sero-survey, respectively. In the remote islands, the seroprevalence was 0.0% (0/144, 95% CI: 0.0-2.5) and 1.6% (2/123, 0.2-5.8) at the 2nd and 3rd survey, respectively. The overall case detection ratios at the 3rd survey were 2.7 (95% CI: 1.3-5.3) in the main island and 2.8 (0.7-11.1) in the remote islands. The highest age-specific case detection ratio was observed in people aged 20-29 years (8.3, 95% CI: 3.3-21.4) in the main island and in those aged 50-59 years (14.1, 2.1-92.7) in the remote islands. The low seroprevalence at the latest survey suggested that a large-scale epidemic had not yet occurred in Okinawa by February 2021. The case detection ratios imply that the cumulative number of incident cases in Okinawa should be 2-3 times higher than that reported by routine surveillance. The ratio was particularly high in young people probably due to a frequent asymptomatic/mild COVID-19 disease in this age group. To accurately measure the scale of the COVID-19 epidemic, it is crucially important to conduct a sero-survey targeting the young.

Climate warming threatens marine life by increasing metabolic oxygen demand while decreasing oxygen availability. Tropical species living in warm, low oxygen environments may be most at risk, but their tolerances and exposures to these stressors remain poorly documented. We evaluated habitat restrictions for two brittle star species from Caribbean coral reefs by integrating field observations, laboratory experiments and an ecophysiological model. The absence of one species from the warmest reefs results from vital activity restrictions during episodic low oxygen extremes, even though average conditions are well within physiological tolerance limits. Over the past decade, warmer temperatures have been significantly correlated with a greater frequency and intensity of hypoxic events. Continued warming will progressively exclude hypoxia-tolerant species, even if average oxygen remains constant. A warming-driven increase in frequency or intensity of low oxygen extremes could similarly accelerate habitat loss across other marine ecosystems.

Marine Protected Areas (MPAs) are widely used as place-based protective measures for restoring and safeguarding marine biodiversity. When ecological connectivity is taken into account during design and management, the results can lead to more effective and resilient MPAs and MPA networks. This publication provides 13 ‘Rules of Thumb’ to support more consistent efforts by MPA managers and marine conservation professionals to implement connectivity conservation and better measure progress towards global conservation targets. These purpose-built tools are intended to inform more effective management and protection of oceans and coasts by covering a diversity of science and policy issues. They can also be used to progress system-based marine conservation as an essential component of national, transboundary, and global policies that establish greater connectivity across borders and at larger scales.

Vesicular stomatitis virus Indiana strain glycoprotein (VSVind.G) mediates broad tissue tropism and efficient cellular uptake. Lentiviral vectors (LVs) are particularly promising, as they can efficiently transduce non-dividing cells and facilitate stable genomic transgene integration; therefore, LVs have an enormous untapped potential for gene therapy applications, but the development of humoral and cell-mediated anti-vector responses may restrict their efficacy. We hypothesized that G proteins from different members of the vesiculovirus genus might allow the generation of a panel of serotypically distinct LV pseudotypes with potential for repeated in vivo administration. We found that mice hyperimmunized with VSVind.G were not transduced to any significant degree following intravenous injection of LVs with VSVind.G envelopes, consistent with the thesis that multiple LV administrations would likely be blunted by an adaptive immune response. Excitingly, bioluminescence imaging studies demonstrated that the VSVind-neutralizing response could be evaded by LV pseudotyped with Piry and, to a lesser extent, Cocal virus glycoproteins. Heterologous dosing regimens using viral vectors and oncolytic viruses with Piry and Cocal envelopes could represent a novel strategy to achieve repeated vector-based interventions, unfettered by pre-existing anti-envelope antibodies. Keywords: VSV, vesiculovirus G, lentivector, immunity, gene delivery, in vivo

Lentiviral vectors (LVs) have been successfully used in clinical trials showing long term therapeutic benefits. Studying the role of cellular proteins in lentivirus HIV-1 life cycle can help understand virus assembly and budding, leading to improvement of LV production for gene therapy. Lentiviral vectors were purified using size exclusion chromatography (SEC). The cellular protein composition of LVs produced by two different methods was compared: the transient transfection system pseudotyped with the VSV-G envelope, currently used in clinical trials, and a stable producer cell system using a non-toxic envelope derived from cat endogenous retrovirus RD114, RDpro. Proteins of LVs purified by size exclusion chromatography were identified by tandem mass spectrometry (MS/MS). A smaller number of cellular protein species were detected in stably produced vectors compared to transiently produced vector samples. This may be due to the presence of co-purified VSV-G vesicles in transiently produced vectors. AHNAK (Desmoyokin) was unique to RDpro-Env vectors. The potential role in LV particle production of selected proteins identified by MS analysis including AHNAK was assessed using shRNA gene knockdown technique. Down-regulation of the selected host proteins AHNAK, ALIX, and TSG101 in vector producer cells did not result in a significant difference in vector production.

We report the first comprehensive review of the industrial process for the catalytic partial oxidation of methanol to formaldehyde using silver catalyst. Formaldehyde remains a key, large volume industrial chemical which is ubiquitous to household, commercial, aviation, pharmaceutical and automotive products due to its high reactivity and versatility. Critical analysis of the various technological approaches used to manufacture formaldehyde, characteristics of silver catalysts employed, process conditions, and performance values has been presented. The three common plant types are almost complete conversion (BASF type); incomplete conversion followed by distillation (ICI type); and the off-gas recycle process which dominates world production due to its extensive use in China. Polycrystalline silver remains the most popular catalyst, with only minimal use of alternate supported catalysts or gauze materials. Catalyst physical properties such as bulk density, purity, mechanical strength, size fraction and sh...

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The evolution of classical ballet from its accepted origins as one method of displaying status and aristocratic power in Renaissance Italy to its Romantic form, featuring professional ballerinas in white costumes dancing en pointe, took place largely during the long eighteenth century. This article discusses this transformation from the dual perspectives of choreography and costume by using the premise that these two vital elements in the presentation of ballet were co-dependent, each prompting the other to develop and evolve. Concentrating on Paris and London, it examines the relationship between court dress, fashion and theatre costume, and how this affected both the choreography and the style of dance throughout the long eighteenth century.

TMEM16F is a Ca2+ -gated ion channel that is required for Ca2+ -activated phosphatidylserine exposure on the surface of many eukaryotic cells. TMEM16F is widely expressed and has roles in platelet activation during blood clotting, bone formation and T cell activation. By combining microscopy and patch clamp recording we demonstrate that activation of TMEM16F by Ca2+ ionophores in Jurkat T cells triggers large-scale surface membrane expansion in parallel with phospholipid scrambling. With continued ionophore application,TMEM16F-expressing cells then undergo extensive shedding of ectosomes. The T cell co-receptor PD-1 is selectively incorporated into ectosomes. This selectivity depends on its transmembrane sequence. Surprisingly, cells lacking TMEM16F not only fail to expand surface membrane in response to elevated cytoplasmic Ca2+, but instead undergo rapid massive endocytosis with PD-1 internalisation. These results establish a new role for TMEM16F as a regulator of Ca2+ activated membrane trafficking.

VSVind.G is currently regarded as the gold-standard envelope glycoprotein to pseudotype lentiviral vectors. However, recently other G proteins derived from vesiculoviruses have been proposed as alternative envelopes. Here, we investigated two commercially available anti-VSVind.G monoclonal antibodies for their ability to cross-react with other vesiculovirus G proteins, identified the epitopes they recognize, and explored their neutralization activity. We have identified 8G5F11, for the first time, as a cross-neutralizing antibody against several vesiculovirus G proteins. Furthermore, we elucidated the two different neutralization mechanisms employed by these two monoclonal antibodies. Understanding how cross-neutralizing antibodies interact with other G proteins may be of interest in the context of host-pathogen interaction and coevolution, as well as providing the opportunity to modify the G proteins and improve G protein-containing medicinal products and vaccine vectors., Vesicular stomatitis virus Indiana strain G protein (VSVind.G) is the most commonly used envelope glycoprotein to pseudotype lentiviral vectors (LV) for experimental and clinical applications. Recently, G proteins derived from other vesiculoviruses (VesG), for example, Cocal virus, have been proposed as alternative LV envelopes with possible advantages over VSVind.G. Well-characterized antibodies that recognize VesG will be useful for vesiculovirus research, development of G protein-containing advanced therapy medicinal products (ATMPs), and deployment of VSVind-based vaccine vectors. Here, we show that one commercially available monoclonal antibody, 8G5F11, binds to and neutralizes G proteins from three strains of VSV, as well as Cocal and Maraba viruses, whereas the other commercially available monoclonal anti-VSVind.G antibody, IE9F9, binds to and neutralizes only VSVind.G. Using a combination of G protein chimeras and site-directed mutations, we mapped the binding epitopes of IE9F9 and 8G5F11 on VSVind.G. IE9F9 binds close to the receptor binding site and competes with soluble low-density lipoprotein receptor (LDLR) for binding to VSVind.G, explaining its mechanism of neutralization. In contrast, 8G5F11 binds close to a region known to undergo conformational changes when the G protein moves to its postfusion structure, and we propose that 8G5F11 cross-neutralizes VesGs by inhibiting this. IMPORTANCE VSVind.G is currently regarded as the gold-standard envelope glycoprotein to pseudotype lentiviral vectors. However, recently other G proteins derived from vesiculoviruses have been proposed as alternative envelopes. Here, we investigated two commercially available anti-VSVind.G monoclonal antibodies for their ability to cross-react with other vesiculovirus G proteins, identified the epitopes they recognize, and explored their neutralization activity. We have identified 8G5F11, for the first time, as a cross-neutralizing antibody against several vesiculovirus G proteins. Furthermore, we elucidated the two different neutralization mechanisms employed by these two monoclonal antibodies. Understanding how cross-neutralizing antibodies interact with other G proteins may be of interest in the context of host-pathogen interaction and coevolution, as well as providing the opportunity to modify the G proteins and improve G protein-containing medicinal products and vaccine vectors.

Retroviral and lentiviral vectors often use the envelope G protein from the vesicular stomatitis virus Indiana strain (VSVind.G). However, lentivector producer cell lines that stably express VSVind.G have not been reported, presumably because of its cytotoxicity, preventing simple scale-up of vector production. Interestingly, we showed that VSVind.G and other vesiculovirus G from the VSV New Jersey strain (VSVnj), Cocal virus (COCV), and Piry virus (PIRYV) could be constitutively expressed and supported lentivector production for up to 10 weeks. All G-enveloped particles were robust, allowing concentration and freeze-thawing. COCV.G and PIRYV.G were resistant to complement inactivation, and, using chimeras between VSVind.G and COCV.G, the determinant for complement inactivation of VSVind.G was mapped to amino acid residues 136–370. Clonal packaging cell lines using COCV.G could be generated; however, during attempts to establish LV producer cells, vector superinfection was observed following the introduction of a lentivector genome. This could be prevented by culturing the cells with the antiviral drug nevirapine. As an alternative countermeasure, we demonstrated that functional lentivectors could be reconstituted by admixing supernatant from stable cells producing unenveloped virus with supernatant containing envelopes harvested from cells stably expressing VSVind.G, COCV.G, or PIRYV.G. Keywords: lentivector, vesiculovirus G, stable packaging

Background: The ability to manipulate the activity of interneurons with optogenetic tools offers the possibility of interfering with diseases caused by altered neuronal inhibition and synchrony, including epilepsy and schizophrenia. To develop vectors for therapeutic approaches, targeting optogenetic constructs to interneurons is therefore a key requirement. We investigated whether the interneuron‐specific promoters glutamic acid decarboxylase (GAD)67 and cholecystokinin (CCK) allowed targeted lentiviral delivery of opsins to interneurons as a whole, or specifically CCK+ interneurons. Methods: We generated lentiviral (LV) plasmids encoding channelrhodopsin (ChR2) and halorhodopsin (NpHR) tagged with fluorophores and driven by GAD67 or CCK promoters. Adeno‐associated virus (AAV) and LV vectors carrying opsins driven by pyramidal cell promoters were used as controls. We transduced neuronal cultures and rodent brain in vivo, immunostained specimens 6–8 weeks after in vivo injection and 7–14 days after in vitro transduction, and evaluated volume and specificity of expression by confocal microscopy. Results: In vitro, 90% (19/21) of LV‐CCK‐NpHR2.0‐EYFP expressing neurons were CCK+. In vivo, LV‐GAD67‐ChR2‐mCherry was expressed in 2.6% (5/193), LV‐GAD67‐NpHR2.0‐EYFP in approximately 15% (43/279) and LV‐CCK‐NpHR2.0‐EYFP in 47% (9/19) of hippocampal GABA+ interneurons. GAD67 vectors expressed in larger volumes than CCK‐driven constructs. AAV vector controls achieved the largest expression volumes. Conclusions: LV‐CCK‐NpHR2.0‐EYFP may be useful for targeting CCK+ interneurons in culture. GAD67/CCK‐driven lentiviral constructs are expressed in vivo, although expression is not specific for interneurons. Overall, expression levels are low compared to opsins driven by pyramidal cell promoters. A better understanding of GAD67 and CCK promoter structure or alternative techniques is required to reliably target opsins to interneurons using viral vectors.

TMEM16F, an ion channel gated by high cytoplasmic Ca2+, is required for cell surface phosphatidylserine exposure during platelet aggregation and T cell activation. Here we demonstrate in Jurkat T cells and HEK293 cells that TMEM16F activation triggers large-scale surface membrane expansion in parallel with lipid scrambling. Following TMEM16F mediated scrambling and surface expansion, cells undergo extensive membrane shedding. The membrane compartment that expands the cell surface does not recycle, nor does it involve endoplasmic reticulum or acidified lysosomes. Surprisingly, T cells lacking TMEM16F expression not only fail to scramble lipid and expand surface membrane, but instead undergo a rapid membrane internalization known as massive endocytosis (MEND) in response to the same Ca2+ stimulus as their parental cells. TMEM16F dependent membrane shedding and MEND traffic similar sets of membrane receptors from the cell surface. The selective participation of the T cell co-receptor PD-1 in this trafficking is determined by its single transmembrane domain, establishing a fundamental role for TMEM16F as a regulator of Ca2+-activated membrane trafficking.

Primary effusion lymphoma (PEL) is a lymphogenic disorder associated with Kaposi's sarcoma-associated herpesvirus (KSHV) infection. Key to the survival and proliferation of PEL is the canonical NF-κB pathway, which becomes constitutively activated following overexpression of the viral oncoprotein KSHV vFLIP (ks-vFLIP). This arises from its capacity to form a complex with the modulatory subunit of the IκB kinase (IKK) kinase, IKKγ (or NEMO), resulting in the overproduction of proteins that promote cellular survival and prevent apoptosis, both of which are important drivers of tumorigenesis. Using a combination of cell-based and biophysical assays together with structural techniques, we showed that the observed resistance to cell death is largely independent of autophagy or major death receptor signaling pathways and demonstrated that direct targeting of the ks-vFLIP–IKKγ interaction both in cells and in vitro can be achieved using IKKγ-mimetic peptides. Our results further reveal that these peptides not only induce cell killing but also potently sensitize PEL to the proapoptotic agents tumor necrosis factor alpha and etoposide and are the first to confirm ks-vFLIP as a tractable target for the treatment of PEL and related disorders. IMPORTANCE KSHV vFLIP (ks-vFLIP) has been shown to have a crucial role in cellular transformation, in which it is vital for the survival and proliferation of primary effusion lymphoma (PEL), an aggressive malignancy associated with infection that is resistant to the majority of chemotherapeutic drugs. It operates via subversion of the canonical NF-κB pathway, which requires a physical interaction between ks-vFLIP and the IKK kinase modulatory subunit IKKγ. While this interaction has been directly linked to protection against apoptosis, it is unclear whether the suppression of other cell death pathways implicated in ks-vFLIP pathogenesis is an additional contributor. We demonstrate that the interaction between ks-vFLIP and IKKγ is pivotal in conferring resistance to apoptosis. Additionally, we show that the ks-vFLIP–IKKγ complex can be disrupted using peptides leading to direct killing and the sensitization of PEL cells to proapoptotic agents. Our studies thus provide a framework for future therapeutic interventions.

Towards (more) complex corpora annotation using a game with a purpose : the case of scientific language. We previously showed that participants in a game with a purpose can produce quality dependency syntax annotations. We present here the results of an experiment aiming at evaluating their production on a more complex corpus of scientific texts on DNA. We precisely describe the complexity of the corpus, then we evaluate the annotations produced by the players as compared to a gold corpus, annotated and adjudicated by experts of the domain., Nous avons précédemment montré qu'il est possible de faire produire des annotations syntaxiques de qualité par des participants à un jeu ayant un but. Nous présentons ici les résultats d'une expérience visant à évaluer leur production sur un corpus plus complexe, en langue de spécialité, en l'occurrence un corpus de textes scientifiques sur l'ADN. Nous déterminons précisément la complexité de ce corpus, puis nous évaluons les annotations en syntaxe de dépendances produites par les joueurs par rapport à une référence mise au point par des experts du domaine.

Expression of the costimulatory receptor 4-1BB is induced by TCR recognition of Ag, whereas 4-1BB ligand (4-1BBL) is highly expressed on activated APC. 4-1BB signaling is particularly important for survival of activated and memory CD8+ T cells. We wished to test whether coexpression of Ag and 4-1BBL by dendritic cells (DC) would be an effective vaccine strategy. Therefore, we constructed lentiviral vectors (LV) coexpressing 4-1BBL and influenza nucleoprotein (NP). Following s.c. immunization of mice, which targets DC, we found superior CD8+ T cell responses against NP and protection from influenza when 4-1BBL was expressed. However, functionally superior CD8+ T cell responses were obtained when two LV were coinjected: one expressing 4-1BBL and the other expressing NP. This surprising result suggested that 4-1BBL is more effective when expressed in trans, acting on adjacent DC. Therefore, we investigated the effect of LV expression of 4-1BBL in mouse DC cultures and observed induced maturation of bystander, untransduced cells. Maturation was blocked by anti–4-1BBL Ab, required cell–cell contact, and did not require the cytoplasmic signaling domain of 4-1BBL. Greater maturation of untransduced cells could be explained by LV expression of 4-1BBL, causing downregulation of 4-1BB. These data suggest that coexpression of 4-1BBL and Ag by vaccine vectors that target DC may not be an optimal strategy. However, 4-1BBL LV immunization activates significant numbers of bystander DC in the draining lymph nodes. Therefore, transactivation by 4-1BBL/4-1BB interaction following DC–DC contact may play a role in the immune response to infection or vaccination.

Purpose/objectives To explore nurses' practice patterns, knowledge, and barriers related to chemotherapy-induced peripheral neuropathy (CIPN). Design Descriptive, cross-sectional. Setting The United States. Sample 408 oncology nurses. Methods A team of eight experts met and developed the CIPN nurse knowledge and preferences survey, which was electronically sent to randomly selected nurses. Main research variables The survey assessed nurses' knowledge and practice patterns regarding assessment strategies and barriers, evidence-based interventions, preferences for education, and perceived gaps in scientific knowledge. Findings Nurses in the survey lacked knowledge regarding neurotoxicity of specific agents and evidence-based treatments. CIPN-focused physical examinations and standardized measurement tools were infrequently used during assessment. The most frequently reported barriers to CIPN assessment included lack of access to measurement tools, lack of specialized skills needed for assessment, lack of confidence, and lack of time. Recommendations for future research included CIPN prevention research, exploration of CIPN-related effects on quality of life, and alternative treatments of CIPN. The majority of participants preferred online educational opportunities. Conclusions Nurses do not consistently integrate evaluation and management of CIPN in their practices. Implications for nursing Educational offerings should incorporate web-based CIPN assessment and management content.

TLR4 interactor with leucine-rich repeats (TRIL) is a brain-enriched accessory protein that is important in TLR3 and TLR4 signaling. In this study, we generated Tril−/− mice and examined TLR responses in vitro and in vivo. We found a role for TRIL in both TLR4 and TLR3 signaling in mixed glial cells, consistent with the high level of expression of TRIL in these cells. We also found that TRIL is a modulator of the innate immune response to LPS challenge and Escherichia coli infection in vivo. Tril−/− mice produce lower levels of multiple proinflammatory cytokines and chemokines specifically within the brain after E. coli and LPS challenge. Collectively, these data uncover TRIL as a mediator of innate immune responses within the brain, where it enhances neuronal cytokine responses to infection.