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Item Diverse Streptococcus pneumoniae Strains Drive a Mucosal-Associated Invariant T-Cell Response Through Major Histocompatibility Complex class I-Related Molecule-Dependent and Cytokine-Driven Pathways.(Oxford University Press, 2018-03-15) Kurioka A; van Wilgenburg B; Javan RR; Hoyle R; van Tonder AJ; Harrold CL; Leng T; Howson LJ; Shepherd D; Cerundolo V; Brueggemann AB; Klenerman PMucosal-associated invariant T (MAIT) cells represent an innate T-cell population that can recognize ligands generated by the microbial riboflavin synthesis pathway, presented via the major histocompatibility complex class I-related molecule (MR1). Streptococcus pneumoniae is a major human pathogen that is also associated with commensal carriage; thus, host control at the mucosal interface is critical. The recognition of pneumococci by MAIT cells has not been defined nor have the genomics and transcriptomics of the riboflavin operon. We observed robust recognition of pneumococci by MAIT cells, using both MR1-dependent and MR1-independent pathways. The pathway used was dependent on the antigen-presenting cell. The riboflavin operon was highly conserved across a range of 571 pneumococci from 39 countries, dating back to 1916, and different versions of the riboflavin operon were also identified in related Streptococcus species. These data indicate an important functional relationship between MAIT cells and pneumococci.Item Bacterial lipopolysaccharide modulates immune response in the colorectal tumor microenvironment.(Nature Portfolio, 2023-08-23) Sulit AK; Daigneault M; Allen-Vercoe E; Silander OK; Hock B; McKenzie J; Pearson J; Frizelle FA; Schmeier S; Purcell RImmune responses can have opposing effects in colorectal cancer (CRC), the balance of which may determine whether a cancer regresses, progresses, or potentially metastasizes. These effects are evident in CRC consensus molecular subtypes (CMS) where both CMS1 and CMS4 contain immune infiltrates yet have opposing prognoses. The microbiome has previously been associated with CRC and immune response in CRC but has largely been ignored in the CRC subtype discussion. We used CMS subtyping on surgical resections from patients and aimed to determine the contributions of the microbiome to the pleiotropic effects evident in immune-infiltrated subtypes. We integrated host gene-expression and meta-transcriptomic data to determine the link between immune characteristics and microbiome contributions in these subtypes and identified lipopolysaccharide (LPS) binding as a potential functional mechanism. We identified candidate bacteria with LPS properties that could affect immune response, and tested the effects of their LPS on cytokine production of peripheral blood mononuclear cells (PBMCs). We focused on Fusobacterium periodonticum and Bacteroides fragilis in CMS1, and Porphyromonas asaccharolytica in CMS4. Treatment of PBMCs with LPS isolated from these bacteria showed that F. periodonticum stimulates cytokine production in PBMCs while both B. fragilis and P. asaccharolytica had an inhibitory effect. Furthermore, LPS from the latter two species can inhibit the immunogenic properties of F. periodonticum LPS when co-incubated with PBMCs. We propose that different microbes in the CRC tumor microenvironment can alter the local immune activity, with important implications for prognosis and treatment response.Item Characterising the Cytokine and Circulating Immune Cell Response After a Single Bout of Eccentric Stepping Exercise in Healthy Untrained Males(Springer Nature, 2023-05-15) Lomiwes D; Barnes M; Shaw O; Ngametua N; Sawyer GM; Burr NS; Miller MRPurpose The mechanisms that underpin exercise-induced muscle damage and recovery are believed to be mediated, in part, by immune cells recruited to the site of injury. The aim of this study was to characterise the effects of muscle damage from bench-stepping on circulating cytokine and immune cell populations post-exercise and during recovery. Methods Ten untrained, healthy male volunteers completed 30 min of bench-stepping exercise to induce muscle damage to the eccentrically exercised leg. Muscle function, muscle pain and soreness were measured before, immediately after and 24, 48 and 72 h after exercise. Plasma creatine kinase, cartilage oligomeric matrix protein, cytokines and circulating immune cell phenotyping were also measured at these timepoints. Results Significant decreases occurred in eccentric, isometric and concentric (P = 0.018, 0.047 and 0.003, respectively) muscle function in eccentrically, but not concentrically, exercised quadriceps post-exercise. Plasma monocyte chemoattractant protein (MCP)-1 concentrations significantly increased immediately after exercise (69.0 ± 5.8 to 89.5 ± 10.0 pg/mL), then declined to below pre-exercise concentrations (58.8 ± 6.3 pg/mL) 72 h after exercise. These changes corresponded with the significant decrease of circulating CD45+ CD16− CD14+ monocytes (5.8% ± 1.5% to 1.9% ± 0.5%; Pre-exercise vs. 48 h) and increase of CD45+ CD3+ CD56− T-cells (60.5% ± 2.2% to 66.1% ± 2.1%; Pre-exercise vs. 72 h) during recovery. Conclusion Bench-stepping induced muscle damage to the quadriceps, which mediated systemic changes in MCP-1, monocytes and T-cells immediately post-exercise and during recovery. Further research is needed to clarify how modulations in immune subpopulations facilitate muscle recovery and adaptation following muscle damage.Item Associations between dietary patterns and an array of inflammation biomarkers and plasma lipid profile in postmenopausal women.(BioMed Central, 2023-05-12) Ilesanmi-Oyelere BL; Kruger MCOBJECTIVE AND DESIGN: In this cross-sectional study, evaluation of the association between four dietary patterns, nutrients and food intakes and an array of systemic inflammation biomarkers and lipid profile among 80 New Zealand postmenopausal women were conducted. MATERIALS: Eighty postmenopausal women participated in the study. A validated food frequency questionnaire was used to collect nutrients and food intake. Four dietary patterns were identified by principal component analysis (PCA) and plasma samples collected for inflammatory biomarkers and lipid profile measures. RESULTS: There were negative correlations between intake of dietary fibre, soluble and insoluble non-starch polysaccharides (NSP), vitamin C and niacin and with almost all the inflammatory markers for the whole group. Vegetables, tea/coffee and especially fruit intake were negatively correlated with the inflammatory biomarkers in the whole group. A high intake of Pattern 1 (potato, bread, and fruit pattern) was associated with a low risk of high interferon (IFN)-α2, IFN-λ, interleukin (IL)-6 and IL-8 levels while a high intake of Pattern 3 (fast-food pattern) was associated high risk of IFN-α2 levels. Multiple linear regression showed a negative correlation between Pattern 2 (soups and vegetables pattern) and levels of C-reactive protein (CRP) as well as ferritin. A positive association was observed between Pattern 3 (fast-food pattern) and CRP levels. Positive correlation was also observed between Pattern 2 and high-density lipoprotein (HDL) and total cholesterol (TC) levels, Pattern 4 (meat and vegetables pattern) was however negatively correlated with TC, low-density lipoprotein (LDL) and TC/HDL ratio. CONCLUSIONS: The result of this study reinforces the contribution and role of diet in modifying inflammation in postmenopausal women.Item Astragalosides Supplementation Enhances Intrinsic Muscle Repair Capacity Following Eccentric Exercise-Induced Injury(MDPI (Basel, Switzerland), 2022-10) Yeh T-S; Lei T-H; Barnes MJ; Zhang LAstragalosides have been shown to enhance endurance exercise capacity in vivo and promote muscular hypertrophy in vitro. However, it remains unknown whether astragalosides supplementation can alter inflammatory response and enhance muscle recovery after damage in humans. We therefore aimed to evaluate the effect of astragalosides supplementation on muscle's intrinsic capacity to regenerate and repair itself after exercise-induced damage. Using a randomized double-blind placebo-controlled cross-over design, eleven male participants underwent 7 days of astragalosides supplementation (in total containing 4 mg of astragalosides per day) or a placebo control, following an eccentric exercise protocol. Serum blood samples and variables related to muscle function were collected prior to and immediately following the muscle damage protocol and also at 2 h, and 1, 2, 3, 5, and 7 days of the recovery period, to assess the pro-inflammatory cytokine response, the secretion of muscle regenerative factors, and muscular strength. Astragalosides supplementation reduced biomarkers of skeletal muscle damage (serum CK, LDH, and Mb), when compared to the placebo, at 1, 2, and 3 days following the muscle damage protocol. Astragalosides supplementation suppressed the secretion of IL-6 and TNF-α, whilst increasing the release of IGF-1 during the initial stages of muscle recovery. Furthermore, following astragaloside supplementation, muscular strength returned to baseline 2 days earlier than the placebo. Astragalosides supplementation shortens the duration of inflammation, enhances the regeneration process and restores muscle strength following eccentric exercise-induced injury.Item Effect of curcumin supplementation on exercise-induced muscle damage: a narrative review(Springer-Verlag GmbH Germany, part of Springer Nature, 2022-07-13) Nanavati K; Rutherfurd-Markwick K; Lee SJ; Bishop NC; Ali ACurcumin, a natural polyphenol extracted from turmeric, is a potent antioxidant and anti-inflammatory agent. In the past few decades, curcumin's ability to impact chronic inflammatory conditions such as metabolic syndrome, arthritis, and cancer has been widely researched, along with growing interest in understanding its role in exercise-induced muscle damage (EIMD). EIMD impacts individuals differently depending on the type (resistance exercise, high-intensity interval training, and running), intensity, and duration of the exercise. Exercise disrupts the muscles' ultrastructure, raises inflammatory cytokine levels, and can cause swelling in the affected limb, a reduction in range of motion (ROM), and a reduction in muscular force-producing capacity. This review focuses on the metabolism, pharmacokinetics of various brands of curcumin supplements, and the effect of curcumin supplementation on EIMD regarding muscle soreness, activity of creatine kinase (CK), and production of inflammatory markers. Curcumin supplementation in the dose range of 90-5000 mg/day can decrease the subjective perception of muscle pain intensity, increase antioxidant capacity, and reduce CK activity, which reduces muscle damage when consumed close to exercise. Consumption of curcumin also improves muscle performance and has an anti-inflammatory effect, downregulating the production of pro-inflammatory cytokines, including TNF-α, IL-6, and IL-8. Curcumin may also improve oxidative capacity without hampering training adaptations in untrained and recreationally active individuals. The optimal curcumin dose to ameliorate EIMD is challenging to assess as its effect depends on the curcumin concentration in the supplement and its bioavailability.Item Serum biomarkers of neuroinflammation and blood-brain barrier leakage in amyotrophic lateral sclerosis(BioMed Central Ltd, 2022-12) Cao MC; Cawston EE; Chen G; Brooks C; Douwes J; McLean D; Graham ES; Dragunow M; Scotter ELAmyotrophic lateral sclerosis (ALS) is an incurable and rapidly progressive neurological disorder. Biomarkers are critical to understanding disease causation, monitoring disease progression and assessing the efficacy of treatments. However, robust peripheral biomarkers are yet to be identified. Neuroinflammation and breakdown of the blood-brain barrier (BBB) are common to familial and sporadic ALS and may produce a unique biomarker signature in peripheral blood. Using cytometric bead array (n = 15 participants per group (ALS or control)) and proteome profiling (n = 6 participants per group (ALS or control)), we assessed a total of 106 serum cytokines, growth factors, and BBB breakdown markers in the serum of control and ALS participants. Further, primary human brain pericytes, which maintain the BBB, were used as a biosensor of inflammation following pre-treatment with ALS serum. Principal components analysis of all proteome profile data showed no clustering of control or ALS sera, and no individual serum proteins met the threshold for statistical difference between ALS and controls (adjusted P values). However, the 20 most changed proteins between control and ALS sera showed a medium effect size (Cohen's d = 0.67) and cluster analysis of their levels together identified three sample subsets; control-only, mixed control-ALS, and ALS-only. These 20 proteins were predominantly pro-angiogenic and growth factors, including fractalkine, BDNF, EGF, PDGF, Dkk-1, MIF and angiopoietin-2. S100β, a protein highly concentrated in glial cells and therefore a marker of BBB leakage when found in blood, was unchanged in ALS serum, suggesting that serum protein profiles were reflective of peripheral rather than CNS biofluids. Finally, primary human brain pericytes remained proliferative and their secretome was unchanged by chronic exposure to ALS serum. Our exploratory study suggests that individual serum cytokine levels may not be robust biomarkers in small studies of ALS, but that larger studies using multiplexed analysis of pro-angiogenic and growth factors may identify a peripheral signature of ALS pathogenesis.Item The responsiveness of the bovine lactoferrin promoter to cytokines and glucocorticoids : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Biochemistry at Massey University(Massey University, 1998) Allen, Kirsty AnnLactoferrin is an iron-binding protein found in many bodily secretions and in the secondary granules of polymorphonuclear leukocytes. While there are many proposed functions for the lactoferrin protein - e.g. for iron storage, antibacterial properties, or a role in inflammation, the specific function(s) of lactoferrin have yet to be elucidated. Evidence that lactoferrin may be involved in inflammation was observed by Harmon et al. (1976) where after the induction of bovine mammary infections, a significant increase in secreted lactoferrin protein was seen during the early phase of the infection. As this increase was during the period of the acute phase response, this suggested that lactoferrin, as was the case with other proteins induced during this time, may have a role in the inflammatory response. The bovine lactoferrin (bLf) promoter contains many putative binding sites for inflammatory modulators, which suggests that the increases in lactoferrin seen during inflammation may be due to activation of lactoferrin gene transcription by these specifically-induced transcription factors. Substantiation of this suggestion would provide further evidence for a specific role for lactoferrin during inflammation. To investigate the cytokine-responsiveness of the bLf promoter, constructs corresponding to various lengths of the putative bLf promoter were linked to the luciferase reporter gene and introduced, by transient transfection, into RL95-2 human endometrial carcinoma cells. Cytokines, glucocorticoids or expression vectors for transcription factors were added to the cells, or potential 'masking' factors in the media such as phenol red or insulin were removed. The luciferase activity of the transfected cells was monitored for significant variation from the basal levels. The addition of cytokines with or without phenol red or insulin did not cause any significant changes in bLF promoter activity. In phenol red-free media, increases in luciferase reporter gene activity were observed after the co-transfection of an expression vector for NF-IL6, the addition of dexamethasone and also the addition of dexamethasone together with the co-transfection of a glucocorticoid receptor expression vector. These data provided evidence that lactoferrin transcription may be induced by inflammatory factors which support the suggestion that lactoferrin has a role in the inflammation process.Item The development of an assay for evaluating the expression of human interleukin-10 parameter region gene linked to inflammatory bowel disease and is application in turmeric assessment : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Human Nutrition at Massey University, Manawatu, New Zealand(Massey University, 2013) Men, XuejingInflammatory bowel disease (IBD) appears in two forms, Crohn's disease (CD) and ulcerative colitis (UC), which are debilitating diseases with less than satisfactory treatments. Despite years of study, the aetiology of this chronic inflammation remains unclear. Evidence from epidemiological and clinical studies supports that it is a complex interaction among environmental, genetic and immune-regulatory factors. Therefore, gene-nutrition based approaches are suggested to be an appropriate candidate in the future prevention and treatment of IBD. Different geographic and racial prevalence of IBD are observed in many epidemiological studies, with highest rates found in developed countries and in Caucasian populations. However, the prevalence has increased dramatically in traditional low-incidence areas during the last two decades, and the racial gap is also closing, indicating that both environmental factors such as diet and genetic predispositions contribute to the IBD susceptibility. The imbalance between pro- and anti-inflammatory cytokines is known to be the key contributor of IBD pathogenesis. Interleukin-10 (IL-10), an anti-inflammatory cytokine, is expressed in many different cells of the adaptive and innate immune system including T regulatory cells, activated macrophages, B regulatory lymphocytes and many other cell types. It plays important part in the regulation of immune response, as was demonstrated in spontaneous colitis in IL-10 deficient mice models, thereforeIL-10 is crucial in the IBD pathogenesis. Three single nucleotide polymorphisms (SNPs) in the promoter region of IL-10 gene, -1082 G/A, -819 C/T and -592 C/A, have been identified to related to IL-10 production and IBD susceptibility, with -1082 G/A as the most relevant SNP.In this research study, Ideveloped a cell-based luciferase reporter assay in which the reporter expression is investigated under the control of promoter containing the variants of interest. Turmeric has a long historical use in Asian medicine for treatment of various diseases. It was shown to exert strong anti-inflammatory effectthrough multiple molecular targets and mechanisms of action. In the second part of my research study, I tested turmeric samples for its ability to alter IL-10 production in the risk polymorphic variant, using the developed assay. The results suggest that curcumin, the bioactive component of turmeric, has the ability to increase IL-10 transcription in the low-producer (ACC)haplotype. The in vitro model of IL-10 promoter assay established in this studyis a novel and valuable tool in assessing IL-10 production at transcriptional level.Furthermore, it provides the possibility of high-throughput screening of food to overcome the functional change of SNPs that are important in human IBD.Item The development of RNA extraction protocols to examine the effects of early exercise on gene expression in the articular cartilage and subchondral bone of Perendale sheep : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science at Massey University, Palmerston North, New Zealand(Massey University, 2010) Pedley, Renée Marie; Pedley, Renée MarieTo examine gene expression in vivo, total RNA was extracted from articular cartilage and subchondral bone. As limited methodology existed for ovine, RNA extraction was performed by optimization of previously published protocols used for other species and in vitro studies (Chomczynski & Mackey, 1995; Chomczynski & Sacchi, 1987; Heinrichs et al., 1997). Reverse transcription polymerase chain reaction (RT-PCR) was used to amplify the extracted RNA to evaluate the gene expression of inflammatory cytokines, collagens, collagenase and housekeeping genes glyceraldehyde phosphate dehydrogenase and Beta-Actin. We observed changes in the expression of inflammatory cytokines, collagen and collagenase genes between exercised and unexercised sheep. The results of this research are consistent with clinical imaging and microscopy studies which suggest that moderate exercise during early life can stimulate an adaptive response in articular cartilage and subchondral bone (Brama, Tekoppele, Bank, Barneveld, & van Weeren, 2000; Firth, 2006; Firth & Rogers, 2005b; Lammi et al., 1993). These changes can have a chondro-protective effect (Jones, Bennell, & Cicuttini, 2003; Otterness et al., 1998) and may reduce susceptibility to athletic injury in later life. Future research using fluorescent probes and polymerase chain reaction may permit quantification of gene expression in real-time to determine the anabolic and catabolic response of articular cartilage with developmental age and exercise in vivo.