Investigation into the role of La/SS-B in Interferon regulation and its relevance in health and human disease

The La/SS-B protein is well documented as an autoantigen for autoimmune conditions such as systemic lupus erythematosus (SLE). Recently, a link between La and viruses has been reported, an interesting observation given the fact that viral infection is a significant risk factor in SLE. However, the direct mechanism by which La functions in the innate immune response to viral challenge remains elusive. As such, we sought to investigate whether La directly regulates the induction of the anti-viral cytokines, type I Interferons, (IFNs) by direct modulation of innate immune toll-like receptor (TLR) or RIG-I-like receptor (RLR) signalling pathways. Our findings have highlighted a dual and complex role for La in the regulation of IFN production. Initial studies demonstrated a role for La in the specific attenuation of IFN promoter transcriptional activity downstream of RLR activation. In confirmation of this, elevated IFN levels were observed following La knockdown in resting cells, compared with controls. Studies in a cohort of SLE patients supported these findings, with decreased La expression observed in SLE patients compared with healthy controls in basal cells, both at an mRNA and protein level. However upon activation of the anti-viral RIG-I receptor, La was shown to promote anti-viral responses by direct interaction with RIG-I receptor, enhancing complex formation between RIG-I and its ligand as well as downstream adaptor protein IPS-1, the overall consequence of which was increased IFN induction. Sendai virus infection experiments supported these findings with depletion of La leading to increased viral infection efficiency and decreased IFN and IFN stimulated gene (ISG) expression. Thus a dual role exists for La in the context of RIG-I-mediated IFN production, with the protein capable of both inhibiting basal type I IFN in resting cells, most likely in an attempt to prevent inappropriate IFN induction and maintain innate immune balance. In the context of infection, however, La is a potent activator of IFN responses, promoting RIG-I binding to its ligand and enhanced type I and type III IFN production, in an effort to protect the host by limiting viral replication and promoting the clearance of the pathogen. Interestingly, we have observed that phosphorylation of La by casein kinase 2 (CK2) may be the regulatory switch that turns La from being active to inactive, and that in cells treated with a CK2-specific inhibitor, La has a higher affinity for RIG-I. With respect to a potential involvement of this protein in SLE we have observed that a higher ratio of expression of a potentially active form of La (neo-La) is observed in SLE patients, suggesting that this imbalance between full length and neo-La in SLE patients contributes to the dysregulated IFN expression observed in patients. Our work has important implications for our understanding of pathways regulating IFN production in the context of SLE

Characterisation of the Upper Respiratory Tract Virome of Feedlot Cattle and its Association with Bovine Respiratory Disease

Bovine respiratory disease (BRD) is a major health problem within the global cattle industry. This disease has a complex aetiology, with viruses playing an integral role. In this study, metagenomics was used to sequence viral nucleic acids in the nasal swabs of BRD affected cattle. Viruses detected included those well known for their association with BRD in Australia (bovine viral diarrhea virus 1), as well as viruses known to be present but not fully characterised (bovine coronavirus) and viruses that have not been reported in BRD affect cattle in Australia (bovine rhinitis, bovine influenza D, and bovine nidovirus). Nasal swabs from a case control study were subsequently tested for 10 viruses and the presence of at least one virus was found to be significantly associated with BRD. Some of the more recently detected viruses had inconsistent association with BRD. Full genome sequences for bovine coronavirus, a virus increasingly associated with BRD, and bovine nidovirus were complete. Both viruses belong to the Coronaviridae family, which are frequently associated with disease in mammals. This study has provided greater insights into the viral pathogens associated with BRD and highlighted the need for further studies to elucidate more precisely the roles viruses play in BRD

Characterisation of the Upper Respiratory Tract Virome of Feedlot Cattle and its Association with Bovine Respiratory Disease

Bovine respiratory disease (BRD) is a major health problem within the global cattle industry. This disease has a complex aetiology, with viruses playing an integral role. In this study, metagenomics was used to sequence viral nucleic acids in the nasal swabs of BRD affected cattle. Viruses detected included those well known for their association with BRD in Australia (bovine viral diarrhea virus 1), as well as viruses known to be present but not fully characterised (bovine coronavirus) and viruses that have not been reported in BRD affect cattle in Australia (bovine rhinitis, bovine influenza D, and bovine nidovirus). Nasal swabs from a case control study were subsequently tested for 10 viruses and the presence of at least one virus was found to be significantly associated with BRD. Some of the more recently detected viruses had inconsistent association with BRD. Full genome sequences for bovine coronavirus, a virus increasingly associated with BRD, and bovine nidovirus were complete. Both viruses belong to the Coronaviridae family, which are frequently associated with disease in mammals. This study has provided greater insights into the viral pathogens associated with BRD and highlighted the need for further studies to elucidate more precisely the roles viruses play in BRD

Epidemiological Risk Factors and Modelling Approaches for Risk Assessment of Lumpy Skin Disease Virus Introduction and Spread: Methodological Review and Implications for Risk-Based Surveillance in Australia

Lumpy skin disease (LSD) is a vector-borne infection caused by the poxvirus lumpy skin disease virus (LSDV) and is a serious disease of cattle, water buffalo, and banteng. While the disease has never occurred in Australia, it is regarded as a growing threat to the Australian cattle industry as there is on-going spread of the disease throughout Asia. The development of geospatial decision support tools, such as spatial epidemiological modelling, may assist in assessing areas at greater risk of this threat. To guide the design of disease modelling approaches to support future risk-based surveillance, existing LSDV epidemiological models need to be evaluated. In this study, we performed a literature review to evaluate existing LSDV epidemiological models, identify key risk factors for introduction and spread of LSDV, and consider previously adopted control strategies. The PRISMA guidelines were used to establish the processes for article selection and information extraction, and the PICO process was used to formulate search terms. From studies that met our inclusion criteria, we extracted information on LSDV epidemiological model structure and parameterisation, risk factors for LSDV transmission and spread, and biosecurity control strategies. The literature search retrieved a total of 402 articles from four databases, of which 68 were identified for inclusion in this review following screening. Of the 68 articles reviewed, 47 explored risk factors associated with LSDV transmission and spread, four explored risk factors of LSDV introduction, four explored existing surveillance strategies in LSD-free countries, and 14 presented epidemiological models. Our findings indicate that there are various risk factors for LSDV transmission in LSD endemic countries, including long-distance airborne movement of infected vectors such as stable flies and cattle movement between countries over land borders. Key risk factors for LSDV spread in LSD endemic countries include physical environmental characteristics, weather conditions, and population distributions of livestock and vectors. Our results indicate that while a variety of modelling studies have been conducted, the majority of studies experimentally explored LSD transmission mechanisms in vectors and cattle. Spatial and spatio-temporal models have primarily been developed for LSD endemic countries and focus on the spread of the disease in terms of environmental factors in relation to previous LSD events. There were very few studies on LSD-free countries, and these only focussed on risk of LSD introduction through specific entry pathways. This review did not identify any literature exploring the risk of spread of LSDV following introduction in LSD-free countries or geospatial modelling of the suitability of LSD-free countries for LSDV incursions. In conjunction with the risk parameters and models described in the identified literature, there is need to consider a wide range of risk factors specific to Australia to inform the design of risk-based surveillance for LSD in Australia

Characterisation of the Upper Respiratory Tract Virome of Feedlot Cattle and Its Association with Bovine Respiratory Disease

Bovine respiratory disease (BRD) is a major health problem within the global cattle industry. This disease has a complex aetiology, with viruses playing an integral role. In this study, metagenomics was used to sequence viral nucleic acids in the nasal swabs of BRD-affected cattle. The viruses detected included those that are well known for their association with BRD in Australia (bovine viral diarrhoea virus 1), as well as viruses known to be present but not fully characterised (bovine coronavirus) and viruses that have not been reported in BRD-affected cattle in Australia (bovine rhinitis, bovine influenza D, and bovine nidovirus). The nasal swabs from a case–control study were subsequently tested for 10 viruses, and the presence of at least one virus was found to be significantly associated with BRD. Some of the more recently detected viruses had inconsistent associations with BRD. Full genome sequences for bovine coronavirus, a virus increasingly associated with BRD, and bovine nidovirus were completed. Both viruses belong to the Coronaviridae family, which are frequently associated with disease in mammals. This study has provided greater insights into the viral pathogens associated with BRD and highlighted the need for further studies to more precisely elucidate the roles viruses play in BRD

Detection methods for common problems in solar home systems

Gemstone Team SHINE (Students Helping to Implement Natural Energy)Solar power is a valuable source of electricity for users in the developing world, yet many solar home systems are working at marginal capacity or not functioning at all. This study has two purposes: (1) to determine how common problems afflicting these systems affect the voltage output, and (2) to use patterns in voltage as a means of detecting these problems, via diagnostic tools. Team SHINE simulated common problems on experimental systems, collecting voltage data from the batteries and panels. Using these data, we created computer algorithms to detect the problem affecting the system. After testing several detection methods, it was found that the most successful performed at 86.5% accuracy. The algorithms can be used in future research to create a device for detecting these problems, allowing them to be addressed earlier

Genetics of SLE: Functional Relevance for Monocytes/Macrophages in Disease

Genetic studies in the last 5 years have greatly facilitated our understanding of how the dysregulation of diverse components of the innate immune system contributes to pathophysiology of SLE. A role for macrophages in the pathogenesis of SLE was first proposed as early as the 1980s following the discovery that SLE macrophages were defective in their ability to clear apoptotic cell debris, thus prolonging exposure of potential autoantigens to the adaptive immune response. More recently, there is an emerging appreciation of the contribution both monocytes and macrophages play in orchestrating immune responses with perturbations in their activation or regulation leading to immune dysregulation. This paper will focus on understanding the relevance of genes identified as being associated with innate immune function of monocytes and macrophages and development of SLE, particularly with respect to their role in (1) immune complex (IC) recognition and clearance, (2) nucleic acid recognition via toll-like receptors (TLRs) and downstream signalling, and (3) interferon signalling. Particular attention will be paid to the functional consequences these genetic associations have for disease susceptibility or pathogenesis

Human Bone Marrow Organoids for Disease Modeling, Discovery, and Validation of Therapeutic Targets in Hematologic Malignancies

A lack of models that recapitulate the complexity of human bone marrow has hampered mechanistic studies of normal and malignant hematopoiesis and the validation of novel therapies. Here, we describe a step-wise, directed-differentiation protocol in which organoids are generated from induced pluripotent stem cells committed to mesenchymal, endothelial, and hematopoietic lineages. These 3D structures capture key features of human bone marrow— stroma, lumen-forming sinusoids, and myeloid cells including proplatelet-forming megakaryocytes. The organoids supported the engraftment and survival of cells from patients with blood malignancies, including cancer types notoriously difficult to maintain ex vivo. Fibrosis of the organoid occurred following TGFβ stimulation and engraftment with myelofibrosis but not healthy donor–derived cells, validating this platform as a powerful tool for studies of malignant cells and their interactions within a human bone marrow–like milieu. This enabling technology is likely to accelerate the discovery and prioritization of novel targets for bone marrow disorders and blood cancers. SIGNIFICANCE: We present a human bone marrow organoid that supports the growth of primary cells from patients with myeloid and lymphoid blood cancers. This model allows for mechanistic studies of blood cancers in the context of their microenvironment and provides a much-needed ex vivo tool for the prioritization of new therapeutics.</p

Interferon Regulatory Factor 8 Regulates Pathways for Antigen Presentation in Myeloid Cells and during Tuberculosis

IRF8 (Interferon Regulatory Factor 8) plays an important role in defenses against intracellular pathogens, including several aspects of myeloid cells function. It is required for ontogeny and maturation of macrophages and dendritic cells, for activation of anti-microbial defenses, and for production of the Th1-polarizing cytokine interleukin-12 (IL-12) in response to interferon gamma (IFNγ) and protection against infection with Mycobacterium tuberculosis. The transcriptional programs and cellular pathways that are regulated by IRF8 in response to IFNγ and that are important for defenses against M. tuberculosis are poorly understood. These were investigated by transcript profiling and chromatin immunoprecipitation on microarrays (ChIP-chip). Studies in primary macrophages identified 368 genes that are regulated by IRF8 in response to IFNγ/CpG and that behave as stably segregating expression signatures (eQTLs) in F2 mice fixed for a wild-type or mutant allele at IRF8. A total of 319 IRF8 binding sites were identified on promoters genome-wide (ChIP-chip) in macrophages treated with IFNγ/CpG, defining a functional G/AGAAnTGAAA motif. An analysis of the genes bearing a functional IRF8 binding site, and showing regulation by IFNγ/CpG in macrophages and/or in M. tuberculosis-infected lungs, revealed a striking enrichment for the pathways of antigen processing and presentation, including multiple structural and enzymatic components of the Class I and Class II MHC (major histocompatibility complex) antigen presentation machinery. Also significantly enriched as IRF8 targets are the group of endomembrane- and phagosome-associated small GTPases of the IRG (immunity-related GTPases) and GBP (guanylate binding proteins) families. These results identify IRF8 as a key regulator of early response pathways in myeloid cells, including phagosome maturation, antigen processing, and antigen presentation by myeloid cells

I care by...

The Care research group at the Royal College of Art (RCA) was conceived in the last week of June 2020, a month after the killing of George Floyd by police in Minnesota, an act which catalysed global protests on systemic racism and police brutality. In the UK, tens of thousands of protesters took to the streets to show solidarity with demonstrators in the US. Coinciding with the easing of the lockdown restrictions imposed to manage the coronavirus, the marches shone a light on the government’s failure to protect Black, Asian and Minority Ethnic people from the disproportionate risk posed by COVID- 19, and on the police’s increased use of stop and search in areas with large BAME populations. The pandemic has shone the harshest of lights on the question of care in the age of neoliberalism: who gets it; who needs it; who does it; who controls it. The Care research group, comprising staff and postgraduate researchers within the School of Arts and Humanities at the RCA, works in this light. Over the course of a year, as the inequalities of the virus were becoming all too clear, the group regularly came together via Zoom to reflect on: the question of how to care for the human body in the technical-patriarchal societies the virus has re-inscribed; the ‘un-doing’ of what Judith Butler describes as the binary of vulnerability and resistance; the politically-transformative potential of prioritising care (rooted in empathy, solidarity, kinship) over capitalist gain; the activation of creative research practices (including but by no means limited to writing, looking, painting, drawing, filming, performing, collecting, assembling, curating, making public) as means of caring/transforming. The group’s activities through the year of trying, failing, and trying again to care for its work and members are gathered in a co-authored Declaration of Care, published here, and expanded upon with attention to some of the methods group members developed in their research through practice. The Declaration was recited in a participatory performance with invited artist Jade Montserrat on 10 March 2021. Over the course of a two-hour webinar, participants including members of the public were invited to draw alongside Montserrat with whatever materials they had to hand as they listened to texts on the vulnerabilities of bodies, the structuring of care within institutions, and the tactile, sensory, healing qualities of creative practice. This book includes a selection of the participants’ drawings, a Reader comprising the texts that were shared, and Montserrat’s drawings created through the performance. Ahead of the performance, Montserrat delivered an address to the Care research group which looked back on a lifetime of calling for a kind of care that was never provided. Excerpts from Montserrat’s address are included here too, alongside a text and image which reflect on the group’s affective reactions to the experience of listening to it, titled Episode. The Declaration is a list of methods (approaches, processes, techniques), an enumeration of how Care research group members have worked, and would like to work: ‘I care by…’. This is a statement which has reverberated throughout the year, which bears repeating, which resounds still. Gemma Blackshaw, Care research group convenor, 2020–202