NIHRDataInsights/HRCSData huggingface dataset:
Title: Dietary polyphenols as modulators of redox signalling pathways to reduce chronic inflammation in the elderly
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The major thiol/disulphide couples: glutathione, thioredoxins (TRx) and cysteine/cystine are key regulators of redox-signalling pathways. These become oxidised during ageing and cause activation of redox-sensitive transcription factors leading to increased pro-inflammatory cytokines. Polyphenols from fruit and vegetables protect against age-related disorders and gene array studies indicate that they modify cellular thiols. The project will determine whether polyphenols and metabolites affect key thiol couples in cell culture and in intervention studies examining subjects of different ages. Lymphocytes from subjects aged 20, 40, 60 and 80+ will be analysed for reduced and oxidised glutathione, cysteine and cystine, nuclear and cytosolic TRx1 and mitochondrial TRx2 redox status, activation of NFkappa-B and cytokine production. Plasma cytokines, reduced and oxidised glutathione, cysteine and cystine will be analysed. Jurkat cells transfected with glutaredoxin1 (Grx1) fused to redox-sensitive green fluorescent protein (roGFP) to monitor sub-cellular reduced/oxidised glutathione couples will be used as a model to identify polyphenols that modify thiol couples. TRx1 and 2 redox couples and cellular cysteine/cystine will also be determined. Activation of redox-sensitive transcription factors and release of cytokines will be examined for those polyphenols found to affect thiol couples. Foodstuffs containing the polyphenols and metabolites effective in the cell model will be studied in 2 day intervention studies in healthy elderly (80+ years) subjects. Urinary metabolites of polyphenols will be measured and lymphocytes analysed for thiol couples, redox-sensitive transcription factors and cytokine production. Subsequently the most effective foodstuff, or combination of foodstuffs, will be studied in a 10 week intervention examining chronic effects on circulating pro-inflammatory cytokines in subjects of different ages.


Research activity codes: 3.3 NUTRITION AND CHEMOPREVENTION
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Understanding and manipulating chemokines and their receptors in the context of adjuvants
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The reasons why certain vaccine adjuvants and/or delivery systems are more or less effective at inducing immune responses or promoting the preferential induction of particular types of response are unknown. However, identifying the factors that determine the induction, magnitude, phenotype and persistence of lymphocyte responses to antigenic challenge will clearly make a significant contribution to the rational design of vaccines. Central to the function of vaccines is the role of adjuvants. These agents are believed to enhance the uptake, transport and availability of antigen to the immune system. We propose that the molecules regulating the movement of the cells of the immune system, namely chemokines and their receptors, orchestrate these processes and that their modulation could enhance adjuvant and consequently vaccine efficacy. Thus understanding how chemokines impact on the induction of adaptive immune responses is a fundamental immunological objective that has significant implications in the management of animal and human health. In particular, this has relevance to use of vaccines in commercial animals and also the ageing human population where vaccines have reduced efficacy.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: RNA processing mechanisms control lymphocyte development and activation
Funder: Biotechnology and Biological Sciences Research Council

Abstract: We propose that post-transcriptional control mediated by RNA binding proteins (RBPs) is a dominant force in the regulation of lymphocyte development and activation. As such it will be important during the ageing process. We will examine this hypothesis using lines of mice with conditional alleles of three zfp36 family members which are inactivated by Cre during T lymphocyte development. Furthermore, we will create mice in which ROD1 and/or PTB function can be regulated at different stages of B and T cell development. We will seek to understand how these RBP are regulated by signal transduction pathways downstream of antigen receptors. For studies of immunity we will employ model antigens and infectious organisms to study the responses of young and old mice. This will be complemented by in vitro studies of lymphocyte differentiation. To identify the specific targets of RBPs, purified naïve and memory lymphocytes will be exposed to UV light to crosslink RNA to protein and specific antibodies used to purify RNA-RBP complexes. The recovered RNA will be identified by deep-sequencing of cDNA. This will be combined with the study of specific splicing patterns using splice junction arrays; the turnover of mRNA using thiouridine labelling methods that allow the labelling of newly synthesized RNA; and the analysis of mRNA association with translating ribosomes. Using informatics approaches we will integrate the knowledge obtained from these experiments on normal lymphocytes with data obtained on RNA handling in lymphocytes in the absence of specific RBPs. These large datasets will be used to generate regulatory maps describing post-transcriptional control. Together, these studies will provide a detailed picture of the global regulation of mRNA handling at the post-transcriptional level in primary cells from animals of different ages.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Reversing senescence and exhaustion signalling pathways in primary human T lymphocytes during ageing
Funder: Biotechnology and Biological Sciences Research Council

Abstract: This study involves the use of freshly isolated leucocyte populations from the peripheral blood of older humans (>70 years). We have specific recruitment programmes for these individuals in place and two research nurses who perform the blood taking. Using cell surface markers, we either investgate specific T cells within mixed populations of leucocytes by flow cytometry or isolate specific T cell subsets by FACS sorting. The functional analyses that will be performed include proliferative assays involving staining with the cell cycle marker ki67, cell death by incorporating propidium iodide and annexin V staining, cytokine production by intracellular cytokine staining and telomerase activity using the telomere repeat amplification protocol (TRAP). We will also introduce specific signalling proteins into primary human T cells by transducing them with VSV vectors to examine directly the role of specific signalling components on functional activity. Our laboratory specializes in the investigation of small populations of T cells. We will investigate varicella zoster specific CD4+ and CD8+ T cell populations by 16 hour stimulation of PBMC with VZV antigens in the presence of brefeldin A, followed by intracellular cytokine staining with IFN-gamma. In individuals who are HLA-DR15 positive, we have access to a class II tetramer and donors who are HLA-A2 we will use class I tetramer both directed against immediate-early 1 protein. We also have MHC class 1 tetramers to immunodominat epitopes of the CMV pp65 protein. We will block senescence signalling using the specific p38 inhibitor BIRB 796, that blocks all isoforms of this kinase. We will block PD-1 signalling by blocking its ligands using a combination of anti-PDL-1 and anti-PDL-2 antibodies.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Identification of endogenous ligands for the Retinoid-related Orphan Receptor gamma
Funder: Biotechnology and Biological Sciences Research Council

Abstract: RORgammat, a nuclear hormone receptor, is important for the development and function of TH17 cells. Although there is some evidence to suggest that oxysterols are natural ligands for RORgammat, the exact endogenous ligands are still unknown. Here, we employ chemical derivatisation combined with liquid chromatography-mass spectrometry (LC-MS) to identify the endogenous ligands of RORgammat. To achieve this we will need to meet three objectives. Objective 1. Quantitative screening of oxysterols during TH cell differentiation under TH1, TH2 and TH17 polarizing conditions to identify the specific oxysterols associated with TH17 cells. This will involve purifying human naïve CD4+ T cells from umbilical cord blood and differentiating the cells in vitro using various combinations of cytokines. The quantitative profile of the oxysterols will be analysed by LC-ms Objective 2. Investigate the interaction of identified oxysterols with RORgammat and their effect on TH17 cell development. The activities of identified oxysterols towards RORgammat will be first tested using biochemical assays. The effect on TH17 cell development and cytokine production will be examined using flow cytometry, RT-PCR and ELISA. The binding of endogenous ligands to RORgammat will be confirmed by immunoprecipation and detection by LC-ms Objective 3. Pharmaceutically and genetically manipulating ligand abundance and investigating their effects on TH17 cell development and function. We will use inhibitors to specifically block the enzymes required to produce ligands to find out their effects on suppression on TH17 cell development and TH17-cytokine production. We will also explore the deactivation pathway of the ligands by over-expressing HSD3B7 and analysing the resultant downstream effects.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: The functional and migratory characteristics of low avidity virus-specific T cells during ageing
Funder: Biotechnology and Biological Sciences Research Council

Abstract: We will investigate freshly isolated leucocyte populations from the blood of young (<40 years) and older humans (>70) as well as lymph node and bone marrow samples paired with blood from healthy individuals. We will either investigate specific T cells within mixed populations of leukocytes by flow cytometry or isolate specific T cell subsets by FACS sorting. Proliferative assays involving staining with the cell cycle marker ki67, cell death by incorporating propidium iodide and annexin V staining, cytotoxicity by degranulation and cytokine production by intracellular cytokine staining will be performed. Fluorochrome labelled MHC class I tetrameric complexes (tetramers) will be used to recognize specific T cells for viral antigens such as cytomegalovirus and Epstein-Barr virus. Furthermore, we will use tetramers that have been engineered to have an altered alpha3 domain in the conserved binding site for CD8 so that only T cells with receptors that bind very avidly to the viral peptide will bind the mutated tetramer. We will use conventional (binds low and high avidity T cells) and mutated tetramers (only binds high avidity cells) containing the same viral peptide to test virus specific cells in the same individual. The clonal diversity of low and high avidity T cells will be compared in freshly isolated T cells and following repeated antigen stimulations by TCR sequence analysis. We will investigate telomere length by fluorescence in situ hybridization coupled to flow cytometry (flow-FISH). We have developed extended our multi-parameter flow-FISH technology to enable the detection of surface and cytoplasmic markers together with telomeres in the same tetramer positive virus-specific T cell. We will use time-lapse microscopy as part of an existing collaboration with Prof. John Greenwood to investigate the migration of T cells across endothelial monolayers. We will also induce mild stress in volunteers using a defined speaking task that was used in dr Riddell's PhD.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Understanding how aggregation influences the immune response to recombinant protein therapeutic drugs
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The overall aim of the proposal is to study the quality and vigour of the immune response to selected model biotherapeutic proteins, and the aggregates generated from them. Our experimental approach will combine studies using a well characterised (BALB/c strain) mouse model of protein immunogenicity and allergenicity, cultured human and mouse dendritic cells (DCs) and human peripheral blood mononuclear cells (PBMC). The programme will comprise the following tasks: Task 1: Production and characterisation of recombinant mouse and human protein aggregates for use in immunological investigations. Task 2: To determine whether aggregation influences the stimulation of adaptive immune responses in vivo. A mouse (BALB/c) model will be used to monitor whether aggregation of recombinant mouse proteins impacts on: a) IgG isotype distribution b) IgE antibody production c) cytokine production (Interleukin [IL]-2, IL-4, IL-6, IL-10, IL-13, IL-17 and IL-22) d) the balance between antigen-driven CD4+ and CD8+ T lymphocyte responses e) the frequency of regulatory T cells (Tregs) f) the number and phenotype of CD11c+ dendritic cells (DC) in the spleen. Task 3: To determine whether aggregation influences the stimulation of primary immune responses in vitro. To address this question, the elicitation of human T and B cell responses in vitro by aggregated and non-aggregated human/humanised proteins will be characterised. Task 4: Characterisation of the mechanistic basis for differential effects of aggregated and non-aggregated proteins to cause immune stimulation. The investigation will determine whether aggregated and non-aggregated proteins display a differential potential to a) provoke cytokine production by peripheral blood mononuclear leukocytes (PBMC). b) drive the activation of mouse and human DC in vitro.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Determining the effects of aging on the splenic marginal zone and the implications for host immunity
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The splenic marginal zone (MZ) is a specialised microenvironment that is important for the the capture and clearance of blood-borne pathogens/antigens. In the MZ a specialized subset of B cells (MZ B cells) is situated to capture blood-borne immune complexes and delivery them to follicular dendritic cells (FDC) in the B-cell follicles. MZ B cells also play vital roles in mounting rapid T-cell-independent (TI) antibody responses to polysaccharide antigens such as those in bacterial capsules, bridging the gap before the induction of high-affinity antibody responses. Thus, the splenic MZ is important for the sampling of blood-borne pathogens/Ag and immunity to systemic infections. We have shown that the MZ microarchitecture is grossly disturbed in aged mice impairing the ability of MZ B cells to deliver immune complexes to FDC. These data reveal an important aging-related defect in the host's ability to sample blood-borne antigens. However, nothing is known of the aging-related factors responsible for the disruption to the MZ microarchitecture, or importantly, how this impacts on immune function, especially TI-antibody responses. Invasive pneumococcal disease is a leading cause of mortality in elderly people, and the efficacy of vaccines against this disease is also decreased. As MZ B-cell deficiencies are associated with elevated risk of pneumococcal disease and poor Ab responses, this suggests that effects of aging on the MZ may likewise impact on the ability of the elderly to mount strong TI-Ab responses. A thorough analysis of the mechanisms that underpin the dramatic aging-related decline in the status of the MZ is crucial to aid our understanding of the factors that influence susceptibility to blood-borne pathogens/antigens, and to improve immunity in the elderly. Therefore, with particular emphasis on MZ B cells, this study will fully characterize the effects of aging on the splenic MZ and identify the molecular and cellular factors which underlie these defects.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Computational systems biology of ageing in haematopoiesis and the immune system
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Ageing is a multidimensional process associated with decline in many physical and psychological functions. These life changes have their origin in our genomes, and are influenced by diet, life-style and the environment. One of the key changes associated with ageing that seriously affects the health of the elderly is a decrease in the functioning of the immune system. The immune system normally works to fight infections to keep us healthy. However, the immune system in the elderly is less active, leaving them much more prone to infection and resulting in deterioration in the quality of life in later years and in many cases early death. In this project we will apply powerful new genome-wide technologies to measure many of the effects of ageing in mouse blood cells, with a particular focus on B cells, the blood cells that make antibodies and form a major part of the immune system. We will use this information to identify specific genes whose expression or function is altered in cells from aged mice, and which may be contributing to a deterioration in immune cell function in the elderly. We will then attempt to change the expression of some of these key ageing-related genes experimentally, to determine their role in acceleration or delay of the ageing process and immune system function. A potential impact of this work is an extension of the healthy life span, by providing new information and understanding of the processes of ageing in immune cells and other cells of the blood.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: New dietary strategies addressing the specific needs of elderly population for an healthy ageing in Europe
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The NU-AGE project will start by designing a new food pyramid for those over 65 years old. This will be developed from food based dietary guidelines used in Europe, illustrating the proportions of different foods that should be included in a balanced diet. The NU-AGE 65+ food pyramid will be designed to meet the nutritional needs of the elderly by emphasising nutrient-density, water, dietary fibre, vitamin D and vitamin B12. To study the effects of the NU-AGE food pyramid on health and ageing factors, elderly citizens across Europe will receive advice, fortified foods and other support to adjust their diets to match the pyramid. Food intake data, blood, urine and other samples will be collected and the results will be compared to those of elderly people not taking part in the dietary intervention. Alongside the dietary intervention, socio-economic determinants for food choice in the elderly will be investigated. In particular at IFR we will investigate the effects of a specific type of diet on adaptive and acquired immunity in elderly individuals. Pre and post intervention analysis of circulating cytokines will be monitored along with more in depth immunological analysis that will give us information of the ability of the diet to modify and possibly potentiate the ability of the aged immune system to cope of infections.


Research activity codes: 3.3 NUTRITION AND CHEMOPREVENTION
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Multifactorial pathways contributing to the development and impact of foot problems in systemic sclerosis (scleroderma)
Funder: Department of Health (England)

Abstract: AIM: The overall aim of this studentship, hosted by the UK's leading clinical rheumatology research unit, is to a) identify and b) address the factors that affect the severity of foot problems in SSc. OBJECTIVES:To achieve this aim we will support the student to achieve three sequential objectives. 1. the impact of foot problems in people with SSc will be quantified. 2. candidate factors that may potentially impact on foot problems will be identified through detailed clinical investigation of clinical findings, combined with state-of-the-art statistical modelling. 3 potentially modifiable factors will be isolated, a pilot intervention will be developed and a proof of concept study undertaken to explore the potential for targeted treatment. METHODS. 1 & 2. A cross-sectional case-control study will explore the impact of foot problems in people with SSc and underpin capture of detailed data representing candidate factors contributing to foot problems in SSc. These will include patient reported outcomes, and clinician assessment of vascular, neurological and musculoskeletal systems in addition to disease-specific measures. The contribution of candidate factors to severity of foot problems will be explored through the development of a Structural Equation Model (SEM).3. Following SEM modelling, potentially modifiable factors will be selected and a simple before and after study will be used to investigate the proof of concept for a targeted therapy addressing those factors making the greatest contribution to the model.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Does use of a prehospital sepsis screening tool by ambulance clinicians increase the number of patients correctly identified as having sepsis in the prehospital environment?
Funder: Department of Health (England)

Abstract: "Sepsis is a life threatening condition claiming 36800 lives each year. Research shows that early diagnosis of sepsis and rapid treatment can significantly reduce the chance of dying. Ambulance clinicians are often the first health care professionals that sepsis patients encounter when they seek help. This project will develop a screening tool to enable ambulance clinicians to make an early diagnosis of sepsis, allowing them to pre-alert the receiving hospital of their imminent arrival. Consequently sepsis patients will receive antibiotic therapy sooner after arriving at hospital potentially leading to improved outcomes and reduced mortality."


Research activity codes: 4.3 INFLUENCES AND IMPACT;4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: INFLAMMATORY AND IMMUNE SYSTEM