Focal Spot, Spring 2009

https://digitalcommons.wustl.edu/focal_spot_archives/1111/thumbnail.jp

In vitro expanded human CD4+CD25+ regulatory T cells suppress effector T cell proliferation.

Regulatory T cells (Tregs) have been shown to be critical in the balance between autoimmunity and tolerance and have been implicated in several human autoimmune diseases. However, the small number of Tregs in peripheral blood limits their therapeutic potential. Therefore, we developed a protocol that would allow for the expansion of Tregs while retaining their suppressive activity. We isolated CD4+CD25 hi cells from human peripheral blood and expanded them in vitro in the presence of anti-CD3 and anti-CD28 magnetic Xcyte Dynabeads and high concentrations of exogenous Interleukin (IL)-2. Tregs were effectively expanded up to 200-fold while maintaining surface expression of CD25 and other markers of Tregs: CD62L, HLA-DR, CCR6, and FOXP3. The expanded Tregs suppressed proliferation and cytokine secretion of responder PBMCs in co-cultures stimulated with anti-CD3 or alloantigen. Treg expansion is a critical first step before consideration of Tregs as a therapeutic intervention in patients with autoimmune or graft-versus-host disease

Association of FCGR3A and FCGR3B haplotypes with rheumatoid arthritis and primary Sjögren's syndrome [POSTER PRESENTATION]

Background Rheumatoid arthritis (RA) is an autoimmune disease that is thought to arise from a complex interaction between multiple genetic factors and environmental triggers. We have previously demonstrated an association between a Fc gamma receptor (FcγR) haplotype and RA in a cross-sectional cohort of RA patients. We have sought to confirm this association in an inception cohort of RA patients and matched controls. We also extended our study to investigate a second autoanti-body associated rheumatic disease, primary Sjögren's syndrome (PSS). Methods The FCGR3A-158F/V and FCGR3B-NA1/NA2 functional polymorphisms were examined for association in an inception cohort of RA patients (n = 448), and a well-characterised PSS cohort (n = 83) from the United Kingdom. Pairwise disequilibrium coefficients (D') were calculated in 267 Blood Service healthy controls. The EHPlus program was used to estimate haplotype frequencies for patients and controls and to determine whether significant linkage disequilibrium was present. A likelihood ratio test is performed to test for differences between the haplotype frequencies in cases and controls. A permutation procedure implemented in this program enabled 1000 permutations to be performed on all haplotype associations to assess significance. Results There was significant linkage disequilibrium between FCGR3A and FCGR3B (D' = -0.445, P = 0.001). There was no significant difference in the FCGR3A or FCGR3B allele or genotype frequencies in the RA or PSS patients compared with controls. However, there was a significant difference in the FCGR3A-FCGR3B haplotype distributions with increased homozygosity for the FCGR3A-FCGR3B 158V-NA2 haplotype in both our inception RA cohort (odds ratio = 2.15, 95% confidence interval = 1.1–4.2 P = 0.027) and PSS (odds ratio = 2.83, 95% confidence interval = 1.0–8.2, P = 0.047) compared with controls. The reference group for these analyses comprised individuals who did not possess a copy of the FCGR3A-FCGR3B 158V-NA2 haplotype. Conclusions We have confirmed our original findings of association between the FCGR3A-FCGR3B 158V-NA2 haplotype and RA in a new inception cohort of RA patients. This suggests that there may be an RA-susceptibility gene at this locus. The significant increased frequency of an identical haplotype in PSS suggests the FcγR genetic locus may contribute to the pathogenesis of diverse autoantibody-mediated rheumatic diseases

Depression as a risk factor for adverse outcomes in coronary heart disease

BACKGROUND: Depression is firmly established as an independent predictor of mortality and cardiac morbidity in patients with coronary heart disease (CHD). However, it has been difficult to determine whether it is a causal risk factor, and whether treatment of depression can improve cardiac outcomes. In addition, research on biobehavioral mechanisms has not yet produced a definitive causal model of the relationship between depression and cardiac outcomes. DISCUSSION: Key challenges in this line of research concern the measurement of depression, the definition and relevance of certain subtypes of depression, the temporal relationship between depression and CHD, underlying biobehavioral mechanisms, and depression treatment efficacy. SUMMARY: This article examines some of the methodological challenges that will have to be overcome in order to determine whether depression should be regarded as a key target of secondary prevention in CHD

Sclerodermatomyositis

The classification of rheumatic diseases is still challenging due to several reasons. First, those diseases have several differential clinical features, which giving overlap symptoms. Second, the etiopathogenesis of those diseases remains elusive.Diagnosis of overlap syndrome is made when there are more than one well-defined connective tissue diseases in one patient, which may develop simultaneously or sequentially.1,2 The prevalence of overlap syndrome among autoimmune diseases is25%.2 The term sclerodermatomyositis or scleromyositisis is used to describe an overlap syndrome in patients with scleroderma and dermatomyositis/polymyositis (DM/PM).2,3,4 Sclerodermatomyositis usually affects adults, and it is rarely found in children.4 The clinical features of this syndrome are myalgia or myositis, arthralgia, scleroderma-like skin changes, Raynaud\u27s phenomenon (RP),2,3 interstitial lung disease, calcinosis,3 mask-like facies, dysphagia or esophageal dysmotility,4 as well as the presence of specific antibody Pm/Scl.2 Skin manifestations as the part of dermatomyositis include periorbital erythema and Gottron\u27s papules.3 We report this case due to its very rare occurrence. According to medical records in the Department of Dermatology as well as Rheumatology at Hasan Sadikin Hospital, Bandung, this is the first case recorded in the last 10 years

Salivary biomarkers : diagnostic potential in oral and systemic diseases in epidemiological surveys

Human saliva is a fluid with many biological functions, and essential for the maintenance of oral health. Several studies report local and systemic biomarkers appearing in saliva, including electrolytes, blood products, enzymes and tissue destruction molecules, inflammatory markers as well as proteins putatively associated with different diseases. However, the clinical utility of salivary diagnostics for the assessment of oral and systemic disease remains elusive. The general aim of this project was to investigate the utility of salivary biomarkers for diagnostic potential of oral- and systemic diseases in large populations. This thesis consists of two different projects: (i). Skåne cohort - A cross-sectional study (Studies I and II): 451 individuals were randomly selected and enrolled for these investigations, including 51% women. All participants were asked to complete a questionnaire, a medical history was taken, a clinical examination was made and stimulated saliva samples were collected. (ii). PAROKRANK sub-cohort (Periodontal disease and the relation to myocardial infarction) (Studies III and IV): A case-control study comprising 400 subjects. In total 200 consecutive patients with a first acute myocardial infarction (AMI) admitted to coronary care units in Sweden from May 2010 to December 2011, and 200 controls without previous AMI, matched for age, gender, residential area were included during the same time period. Dental examinations were performed, blood and stimulated saliva samples were collected eight to ten weeks after the myocardial infarction (MI). Matched controls were examined within one to two weeks after the MI patients. Study I: The aim of this study was to investigate if selected salivary biomarkers could be used for epidemiological studies for detection of periodontitis. Our findings showed that patients with severe periodontitis had elevated salivary concentrations of interleukin (IL) -1β (IL-1β) and matrix metalloproteinase (MMP) -8 (MMP-8), as well as and increased ratio of MMP-8/ tissue inhibitor of metalloproteinase-1 (TIMP-1). Study II: The aim of this study was to investigate if certain salivary biomarkers could be used for detection of common systemic inflammatory diseases. The results of our study showed that salivary IL-8 concentrations were higher in patients with bowel disease and subjects who had experience of tumor diseases. MMP-8 levels were elevated in saliva from patients with diabetes, muscle and joint diseases or previously had undergone cardiac surgery. Study III: The aim of this study was to investigate whether salivary concentrations of selected cardiovascular biomarkers could be used to identify patients with a previous MI and whether such markers, in plasma and saliva, were related to periodontal status. There was no difference between participants with or without MI in regards of N-terminal prohormone of brain natriuretic peptide (NT- pro BNP) and growth differentiation factor-15 (GDF-15) levels in saliva. However, cystatin C, NT- pro BNP and GDC-15 levels were higher in MI patients with other co-morbidities. The levels of cystatin C were lower in saliva from patients with MI. GDF-15 levels correlated with periodontal status in both groups. Further, there was no correlation between plasma and saliva levels. Study IV: The aim of this study was to explore the levels of the inflammatory markers, MMP-8, MMP-9, TIMP-1 and myeloperoxidase (MPO) in saliva with regards to previous MI and periodontal disease. The analyzed biomarkers correlated significantly with each other and most of the periodontal parameters in both study groups. Salivary MMP-8 and MPO levels were significantly higher in non- MI subjects. In conclusion, the findings in this project indicates that certain salivary biomarkers have the potential to be used for screening purposes in epidemiological studies related to both oral and systemic diseases. In addition, the selected salivary biomarkers in our studies could be seen as markers for increased local and systemic inflammation

Immune-mediated mechanisms of (mal)adaptive renal tissue repair

The aim of this thesis is to increase our knowledge into the immunopathogenesis of renal sterile inflammatory diseases, with a focus on the innate immune response activation by the danger protein S100A8/A9 and the receptor TREM-1. We found that S100A8/A9 is a pivotal player in renal repair following hypoxic damage, through an immune-regulatory role resulting in the alternative activation of macrophages. By controlling excessive M2 polarization, S100A8/A9 fine-tunes the adaptive response of the kidney to IR-induced AKI ( Chapter 2 ). Conversely, in chronically inflamed kidneys, S100A8/A9 contributes to the development of fibrosis possibly through a direct effect on dedifferentiation and apoptosis in tubular epithelial cells. This effect was independent of S100A8/A9-induced inflammation and recruitment of leukocytes into the kidney ( Chapter 3 ). Instead, TREM-1 is not involved in the amplification of acute renal damage and inflammation in preclinical model of acute kidney injury. Additionally, renal transplanted patients carrying the TREM1 gene variant p.Thr25Ser do not show any association with pathological consequences after transplantation ( Chapter 5 ). TREM-1, however, limits the maladaptive repair post IR-induced AKI, through a direct effect on tubular epithelial mitochondrial homeostasis and energy production, empowering cell cycle progression. The metabolic advantage provided by TREM-1 is indispensable for tubular proliferation, which sustains renal repair and accelerates the recovery from IR ( Chapter 6 ). In the preclinical model of chronic kidney disease TREM-1 and its adapter molecule DAP12 are not involved in the development of renal fibrosis. DAP12, partly through TREM-1, modulates the renal inflammatory response following unilateral ureteral obstruction ( Chapter 7 )

SYSTEM AND METHOD FOR DETERMINING THE DEGREE OF ABNORMALITY OF A PATIENT'S VITAL SIGNS

A system and method for determining the degree of abnormality of a vital sign of a patient by obtaining the clinical profile of said patient and determining the statistical difference between the vital sign of the patient and the vital signs of previously evaluated patients having similar clinical profiles. The vital signs of previously evaluated patients having similar clinical profiles are determined based on matching the attributes of the patent's clinical profile to the clinical profiles of previously evaluated patients. The statistical difference, and the patent's clinical profile may be exported to an electronic medical record system or printed in hard copy for inclusion in the patient's medial file

Nitric oxide and peroxynitrite in health and disease

The discovery that mammalian cells have the ability to synthesize the free radical nitric oxide (NO) has stimulated an extraordinary impetus for scientific research in all the fields of biology and medicine. Since its early description as an endothelial-derived relaxing factor, NO has emerged as a fundamental signaling device regulating virtually every critical cellular function, as well as a potent mediator of cellular damage in a wide range of conditions. Recent evidence indicates that most of the cytotoxicity attributed to NO is rather due to peroxynitrite, produced from the diffusion-controlled reaction between NO and another free radical, the superoxide anion. Peroxynitrite interacts with lipids, DNA, and proteins via direct oxidative reactions or via indirect, radical-mediated mechanisms. These reactions trigger cellular responses ranging from subtle modulations of cell signaling to overwhelming oxidative injury, committing cells to necrosis or apoptosis. In vivo, peroxynitrite generation represents a crucial pathogenic mechanism in conditions such as stroke, myocardial infarction, chronic heart failure, diabetes, circulatory shock, chronic inflammatory diseases, cancer, and neurodegenerative disorders. Hence, novel pharmacological strategies aimed at removing peroxynitrite might represent powerful therapeutic tools in the future. Evidence supporting these novel roles of NO and peroxynitrite is presented in detail in this review