Tumor necrosis factor alpha (TNF-alpha), a proinflammatory/proatherogenic cytokine, has been implicated as the cause hyperlipidemia in patients with chronic inflammatory joint disease including rheumatoid arthritis, psoriatic arthritis and ankylosing spondylitis. In this study, we have measured the effect of long-term moderate intensity exercise training on blood lipid levels and on the ex vivo production of TNF-alpha by mitogen-stimulated peripheral blood mononuclear cells. Study participants underwent six-months of tri-weekly supervised training sessions, each consisted of a combination of aerobic (52%), resistance (40%), and flexibility (8%) exercises. Forty-three subjects, 25 women average age 49.7 years and 18 men average age 48.1 years, successfully completed the study. Both men and women exercised an average of 2.5 hours per week. Exercise training resulted in a 10% decrease in total cholesterol levels (P=0.0136), an 18% decrease in low density lipoprotein cholesterol levels (P=0.0043), a 20.1% decrease in total cholesterol/high density lipoprotein cholesterol ratios (P=0.0030), and a 51.5% decrease in the production of TNF-alpha (P

In most suppressor T-cell-mediated phenomenon studied so far in the chicken, the suppressor cells appear to be CT8 + CT4 -. This chapter demonstrates the presence of both antigen-specific helper and suppressor T cells in both normal and A+¦ chickens after a single i.v. antigen injection. The effect of cimetidine, type II histamine receptors (H2R) antagonist, on suppression of the secondary anti- sheep red blood cell (SRBC) response, induced by addition of antigen on day 0 of culture, was also investigated. Transfer of bovine serum albumin (BSA)-tolerant spleen cells without or with specifically primed adult spleen cells into normal recipients, followed by BSA challenge, prevented any significant primary or secondary anti-BSA response. The suppression was antigen specific, since BSA-tolerant spleen cells were unable to suppress either primary or secondary responses of B cells to SRBC. Since suppressor cells could not be demonstrated, the T-cell tolerance in these chickens appeared to be primarily due to deletion of antigen-responsive cells.

Although it is recognized that the mechanical stresses associated with physical activity augment bone mineral density and improve bone quality, our understanding of how exercise modulates bone homeostasis at the molecular level is lacking. In a before and after trial involving 43 healthy adults, we measured the effect of six months of supervised exercise training on the spontaneous and phytohemagglutinin-induced production of osteoclastogenic cytokines (interleukin-1α, tumor necrosis factor-α), antiosteoclastogenic cytokines (transforming growth factor-β1 and interleukins 4 and 10), pleiotropic cytokines with variable effects on osteoclastogenesis (interferon-γ, interleukin-6), and T cell growth and differentiation factors (interleukins 2 and 12) by peripheral blood mononuclear cells. We also measured lymphocyte phenotypes and serum markers of bone formation (osteocalcin), bone resorption (C-terminal telopeptides of Type I collagen), and bone homeostasis (25 (OH) vitamin D, estradiol, testosterone, parathyroid hormone, and insulin-like growth factor 1). A combination of aerobic, resistance, and flexibility exercises done on average of 2.5 hours a week attenuated the production of osteoclastogenic cytokines and enhanced the production of antiosteoclastogenic cytokines. These changes were accompanied by a 16% reduction in collagen degradation products and a 9.8% increase in osteocalcin levels. We conclude that long-term moderate intensity exercise exerts a favorable effect on bone resorption by changing the balance between blood mononuclear cells producing osteoclastogenic cytokines and those producing antiosteoclastogenic cytokines. This trial is registered with Clinical Trials.gov Identifier: NCT02765945.

Chronic granulomatous disease (CGD) is associated with defective function of the NADPH-oxidase system in conjunction with phagocytic defects which leads to granuloma formation and serious infectious complications. This is often associated with significant morbidity and mortality. The association of defective phagocyte function with other coincidental immune defects is unknown. Defects in innate pathways seen with CGD, including complement systems, and toll-like and dectin receptor pathways, have not been described before. We present the case of a 2-year old male patient hospitalized with recurrent pneumonia, a non-healing skin ulcer, necrotizing lung granulomas, and epididymo-orchitis. Defective neutrophil chemiluminescence was detected by dihydrorhodamine (DHR) testing. Further evaluation demonstrated characteristic molecular mutations of CYBB consistent with CGD. Immune evaluation demonstrated polyclonal hyperglobulinemia, but a greatly reduced mannose binding lectin (MBL) level. Six biallelic polymorphisms in MBL gene and its promoter were analyzed using Light CyclerTM Real-time PCR assay. The LXPA/LYPB haplotype of MBL was detected in our patient; the latter is the defective haplotype associated with low MBL levels. Due to the implications for innate immunity and the protection against bacterial, viral, and fungal infections provided by MBL, a deficiency of this protein may have disastrous consequences on the long term outcomes of CGD. MBL deficiency can also complicate other disorders affecting the immune system, significantly increasing the risk of infection in such patients. Further studies looking at the frequency and implications of MBL deficiency in CGD are needed.

BACKGROUND:Human mast cells are capable of a wide variety of inflammatory responses and play a vital role in the pathogenesis of inflammatory diseases such as allergy, asthma, and atherosclerosis. We have reported that cigarette smoke extract (CSE) significantly increased IL-6 and IL-8 production in IL-1β-activated human mast cell line (HMC-1). Baicalein (BAI) has anti-inflammatory properties and inhibits IL-1β- and TNF-α-induced inflammatory cytokine production from HMC-1. The goal of the present study was to examine the effect of BAI on IL-6 and IL-8 production from CSE-treated and IL-1β-activated HMC-1.METHODS:Main-stream (Ms) and Side-stream (Ss) cigarette smoke were collected onto fiber filters and extracted in RPMI-1640 medium. Two ml of HMC-1 at 1 × 106 cells/mL were cultured with CSE in the presence or absence of IL-1β (10 ng/mL) for 24 hrs. A group of HMC-1 cells stimulated with both IL-1β (10 ng/ml) and CSE was also treated with BAI. The expression of IL-6 and IL-8 was assessed by ELISA and RT-PCR. NF-κB activation was measured by electrophoretic mobility shift assay (EMSA) and IκBα degradation by Western blot.RESULTS:Both Ms and Ss CSE significantly increased IL-6 and IL-8 production (p < 0.001) in IL-1β-activated HMC-1. CSE increased NF-κB activation and decreased cytoplasmic IκBα proteins in IL-1β-activated HMC-1. BAI (1.8 to 30 μM) significantly inhibited production of IL-6 and IL-8 in a dose-dependent manner in IL-1β-activated HMC-1 with the optimal inhibition concentration at 30 μM, which also significantly inhibited the enhancing effect of CSE on IL-6 and IL-8 production in IL-1β-activated HMC-1. BAI inhibited NF-κB activation and increased cytoplasmic IκBα proteins in CSE-treated and IL-1β-activated HMC-1.CONCLUSIONS:Our results showed that CSE significantly increased inflammatory cytokines IL-6 and IL-8 production in IL-1β-activated HMC-1. It may partially explain why cigarette smoke contributes to lung and cardiovascular diseases. BAI inhibited the production of inflammatory cytokines through inhibition of NF-κB activation and IκBα phosphorylation and degradation. This inhibitory effect of BAI on the expression of inflammatory cytokines induced by CSE suggests its usefulness in the development of novel anti-inflammatory therapies.

Henoch-Schonlein Purpura (HSP) is a small vessel vasculitis mediated by IgA-immune complex deposition. It is characterized by the clinical tetrad of non-thrombocytopenic palpable purpura, abdominal pain, arthritis and renal involvement. Pathologically, it can be considered a form of immune complex-mediated leukocytoclastic vasculitis (LCV) involving the skin and other organs. Though it primarily affects children (over 90% of cases), the occurrence in adults has been rarely reported. Management often involves the use of immunomodulatory or immune-suppressive regimens.

Atherosclerosis is now considered an inflammatory disease, representing the paradigm shift over the last several decades. More intriguing recent data suggest that human inflammatory vasculopathy can by triggered by infectious agents. Even minor infections such as gingivitis have been linked to accelerated vascular disease, while profoundly inflammatory disorders such as rheumatoid arthritis have shown a propensity to increased vascular complications. Human mast cells are often found perivascularly and at mucosal sites and and are of pivotal importance to host defense and the inflammatory response. Recent studies demonstrate that mast cells also play roles in the genesis of atherosclerotic vascular disease. Mast cells express toll-like receptors (TLRs) and also express other cell surface receptors such as the receptor for interleukin-1 (IL-1r). Interactions between bacterial pathogens and mast cell TLR and/or related receptors can lead to the elaboration of inflammatory cytokines. These can recruit/activate other inflammatory cells that further accelerate interactions with adaptive immunity (T and B cells). The elaboration of IL-6 by mast cells can contribute to the hepatic production of C reactive protein (CRP), an established biomarker for atherosclerosis. The elaboration of IL-1 from the mast cell (autocrine) or from adventitial cells (paracrine) can lead to further mast cell activation, resulting in an inflammatory loop characterized by further endocrine IL-6 and inflammatory cytokine production, with resultant recruitment and/or activation of other inflammatory cell types and progression of inflammatory vasculopathy. These effects may be mediated by mast cell expression of nuclear factor kappaB and mitogen activated protein kinases (MAPK). Inhibition of mast cell activation and mediator synthesis may play adjunctive futuristic roles in the management of cardiovascular disease.

Human endothelial cells are multifunctional cells that line blood vessels and are capable of secreting a variety of biologically active mediators. They normally maintain vascular hemostasis and prevent thrombotic complications. When affected by infection, stress, hypertension, dyslipidemia, or high homocysteine levels, endothelial cells undergo changes resulting in "dysfunction," characterized typically by decreased endothelial expression of nitric oxide, enhanced expression of cell adhesion molecules, and associated increased binding of circulating leukocytes to these cells. There is accompanying cytokine and chemokine elaboration, resulting in cellular recruitment and the orchestration of an acute inflammatory response that can culminate in chronic inflammation if reparative mechanisms are not operative. This review will address the basic biology of endothelial cells; the expression and regulation of endothelial-derived cytokines, chemokines, and growth factors; the transcriptional regulation of these genes in endothelial cells; and the role played by these fascinating cells in human disease.

Exercise affects various components of the immune system that could be clinically beneficial or deleterious. Accordingly, moderate exercise stimulates the immune system, while intense exercise suppresses certain immune cell activities. Some of the immune-regulation observed may be due to the regulatory T cells. This could also explain the difference between moderate and strenuous exercisers and the probable reason for lower incidence of autoimmunity in active individuals. Intense exercisers (Trained -regular) and Moderate exercisers (untrained- occasional) were recruited as volunteers from of our university community after IRB approval. Blood samples were collected. PBMCs were used for immune cell profile analysis and the plasma was used for cytokine estimation. Results suggest recruitment of all lymphocyte subpopulations to the vascular compartment: CD4, CD8, B, and Natural killer (NK) cells. Depending on intensity of exercise, the CD4-to-CD8 ratio decreased, reflecting the greater increase in CD8s than CD4s. Both memory and naive phenotype were found, while memory type prevailed. The number of Tregs was increased in all subjects and inversely correlated with IL-6 concentration. Resistant training associated initiation of inflammatory cytokine suggest metabolic link between skeletal muscles and the lymphoid system. It is therefore to be expected that moderate exercise protects against malignancy or infection whereas exhaustive exercise is linked to increased pathology.

Human mast cells play a role in the pathogenesis of inflammatory airway diseases. We have reported that CSE increased IL-6 and IL-8 production in IL-1beta activated human mast cells (HMC-1). Baicalein (BAI) has been shown to have anti-inflammatory properties. This study aimed to examine the effect of BAI on IL-6 and IL-8 production in HMC-1. Two ml of HMC-1 at 1 x 106 cells/ml were cultured either with or without IL-1beta (10 ng/ml), and in the presence or absence of mainstream (Ms) or sidestream (Ss) CSE (0.25 mg/ml or 0.125 mg/ml), and BAI (30 μM). Treated HMC-1 were incubated for 24 hours and culture supernatants were collected and analyzed for the levels of IL-6 and IL-8 by ELISA. Both concentrations of Ms and Ss CSE significantly increased production of both IL-6 (p < 0.0001) and IL-8 (p < 0.0005) in IL-1beta activated HMC-1. BAI (30 μm) decreased IL-6 and IL-8 production in the Ms CSE (0.25 mg/ml) + IL-1beta treated HMC-1 culture from 2615 ± 38 to 251 ± 19 pg/ml (p < 0.0000001) and from 583 ± 37 to 179 ± 7 pg/ml (p < 0.0005), respectively. A similar inhibitory effect of BAI was also observed in the Ss CSE + IL-1beta treated HMC-1 culture. Our results show that BAI inhibits the enhancing effects of CSE on inflammatory cytokine production in IL-1beta activated mast cells. It suggests that BAI may be a novel candidate for treatment of smoking related inflammatory airway disease (Supported by the Ruth R. Harris Endowment and RDC of ETSU).

The paradigm shift in cardiovascular biology has been the understanding that atherosclerosis involves not just a mechanical deposition of lipids in the vessel wall, but a dynamic process involving the inflammatory response with cellular infiltration and inflammatory mediator expression. Typical cellular elements that have been studied include endothelial cells, vascular smooth muscle, T lymphocyte and the macrophage. Recent data suggests a role for the human mast cell. The human mast cell is a tissue-dwelling cell, typically perivascular in distribution. This multifunctional cell responds rapidly to challenge with the release of inflammatory mediators that can orchestrate an immune response and may have relevance to atherogenesis. Mast cells have been shown to modulate various aspects of cardiovascular disease such as atherogenesis (endothelial activation, cytokine generation and foam cell formation) as well as rupture of an unstable atheromatous plaque. Mast cell activation in the context of cardiovascular disease may occurby cognate cell-cell interactions (interactions with macrophages, T cells, endothelial cells or smooth muscle) or by non-cognate means (such as lipoproteins and other pro-atherogenic components). More studies are required in order to better understand the molecular role of mast cells in vascular inflammatory disease.

Mannose-binding lectin (MBL) and the Mannose-binding lectin-associated serine proteases (MASPs) are an essential aspect of innate immune responses that probably play an important but understudied role in cutaneous function. The MBL-MASP pathway appears to exert its primary role by assisting in the clearance of apoptotic skin cells (thus preventing accumulation and a subsequent autoimmune response) and promoting opsonophagocytosis of invading pathogens, limiting their dissemination. Deficiencies of the pathway have been described and are associated with infectious, autoimmune and vascular complications. However, the role of this pathway in dermatological disease is essentially unexplored. We describe 6 patients presenting with recurrent inflammatory and/or infectious skin conditions who also demonstrated severely low MBL levels. One patient also had a defect in the MASP2 gene. Genotype analysis revealed specific point mutations in the MBL2 promoter in all 6 patients and a variant MASP-2 gene in one patient. Five patients presented recurrent pustular skin infections (cellulitis, folliculitis and cutaneous abscess). A case of Grover's disease and one forme fruste of Behcet's syndrome (orogenital ulcers) were also observed. The patients responded to antimicrobial therapy, although in some, recurrence of infection was the rule. It appears that MBL deficiency may contribute to recurrent skin infections and to certain forms of inflammatory skin disease. The mechanisms may relate to the role of this pathway in innate immunity, removal of apoptotic cells and in immune complexes. Further study of MBL pathway defects in dermatological disease is required.

Human endothelial cells are multifunctional cells that line blood vessels and are capable of secreting a variety of biologically active mediators. They normally maintain vascular hemostasis and prevent thrombotic complications. When affected by infection, stress, hypertension, dyslipidemia, or high homocysteine levels, endothelial cells undergo changes resulting in "dysfunction," characterized typically by decreased endothelial expression of nitric oxide, enhanced expression of cell adhesion molecules, and associated increased binding of circulating leukocytes to these cells. There is accompanying cytokine and chemokine elaboration, resulting in cellular recruitment and the orchestration of an acute inflammatory response that can culminate in chronic inflammation if reparative mechanisms are not operative. This review will address the basic biology of endothelial cells; the expression and regulation of endothelial-derived cytokines, chemokines, and growth factors; the transcriptional regulation of these genes in endothelial cells; and the role played by these fascinating cells in human disease.

BACKGROUND:Altered levels of Immunoglobulin E (IgE) represent a dysregulation of IgE synthesis and may be seen in a variety of immunological disorders. The object of this review is to summarize the historical and molecular aspects of IgE synthesis and the disorders associated with dysregulation of IgE production.METHODS:Articles published in Medline/PubMed were searched with the keyword Immunoglobulin E and specific terms such as class switch recombination, deficiency and/or specific disease conditions (atopy, neoplasia, renal disease, myeloma, etc.). The selected papers included reviews, case reports, retrospective reviews and molecular mechanisms. Studies involving both sexes and all ages were included in the analysis.RESULTS:Both very low and elevated levels of IgE may be seen in clinical practice. Major advancements have been made in our understanding of the molecular basis of IgE class switching including roles for T cells, cytokines and T regulatory (or Treg) cells in this process. Dysregulation of this process may result in either elevated IgE levels or IgE deficiency.CONCLUSION:Evaluation of a patient with elevated IgE must involve a detailed differential diagnosis and consideration of various immunological and non-immunological disorders. The use of appropriate tests will allow the correct diagnosis to be made. This can often assist in the development of tailored treatments.

Abstract Human mast cells are multifunctional cells capable of inflammatory responses and associated with allergy, asthma, and atherosclerosis. This study aims to examine effects and mechanisms of CSE on the expression of inflammatory cytokines in mast cells. Main- (Ms) and Side-stream (Ss) cigarette smoke were collected onto fiber filters and extracted in RPMI-1640 medium. Two ml of HMC-1 at 1 x 106 cells / mL were cultured with CSE in the presence or absence of IL-1β (10 ng / mL) for 24 hrs. The expression of IL-6 and IL-8 was assessed by ELISA and RT-PCR. NF-κB activation was measured by electrophoretic mobility shift assay (EMSA) and IκBα degradation by Western blot. Both Ms and Ss CSE significantly increased IL-6 and IL-8 production (p < 0.001) in IL-1β-activated HMC-1. CSE increased NF-κB activation and decreased cytoplasmic IκBα proteins in IL-1β-activated HMC-1. These results suggest that CSE enhance the expression of inflammatory cytokines through increased NF-κB activation and IκBα degradation, which may partially explain why cigarette smoke contributes to lung and cardiovascular diseases. (Supported by the Ruth R. Harris Endowment and RDC of ETSU)

Human mast cells are multifunctional cells capable of a wide variety of inflammatory responses and associated with allergy, asthma, and atherosclerosis. Our previous studies have shown that IL‐1β activates human mast cells to produce selected inflammatory cytokines. We examined effects of cigarette smoke extract (CSE) on the production of inflammatory cytokines from IL‐1β‐activated human mast cells. Mainstream (Ms) and Sidestream (Ss) cigarette smoke were collected onto fiber filters and extracted in RPMI‐1640 medium. Two ml of HMC‐1 at 1 × 106 cells/mL were cultured with Ms and Ss CSE at various concentrations in the presence or absence of IL‐1β (10 ng/mL) for 24 hrs. The supernatants were harvested and assayed for IL‐6, IL‐8, and MCP‐1 by ELISA. The Ms and Ss CSE (32.5 to 500 μg/mL) alone induced either no or trace amounts of cytokines from HMC‐1. However, both Ms and Ss CSE significantly increased IL‐6 and IL‐8 production (all p < 0.001) from IL‐1β‐activated HMC‐1 in a dose dependent fashion, except Ms CSE at 500 μg/mL, which significantly decreased IL‐6 production (p < 0.0001). Contrarily, both Ms and Ss CSE at concentrations from 62.5 to 500 μg/mL, but not 32.5 μg/mL, significantly inhibited MCP‐1 production (all p < 0.05) by IL‐1β‐activated HMC‐1 in a dose dependent fashion. The effects of Ms CSE appeared to be more potent than that of Ss CSE. The results show that CSE had differential effects on the production of inflammatory cytokines from IL‐1β‐activated mast cells, which may partially explain why cigarette smoke contributes to lung and cardiovascular diseases. (Supported by the Ruth R. Harris Endowment and RDC of ETSU)

In Asian countries where the Buddhism and Taoism are mainstream religions, incense burning is a daily practice. A typical composition of stick incense consists of 21% (by weight) of herbal and wood powder, 35% of fragrance material, 11% of adhesive powder, and 33% of bamboo stick. Incense smoke (fumes) contains particulate matter (PM), gas products and many organic compounds. On average, incense burning produces particulates greater than 45 mg/g burned as compared to 10 mg/g burned for cigarettes. The gas products from burning incense include CO, CO2, NO2, SO2, and others. Incense burning also produces volatile organic compounds, such as benzene, toluene, and xylenes, as well as aldehydes and polycyclic aromatic hydrocarbons (PAHs). The air pollution in and around various temples has been documented to have harmful effects on health. When incense smoke pollutants are inhaled, they cause respiratory system dysfunction. Incense smoke is a risk factor for elevated cord blood IgE levels and has been indicated to cause allergic contact dermatitis. Incense smoke also has been associated with neoplasm and extracts of particulate matter from incense smoke are found to be mutagenic in the Ames Salmonella test with TA98 and activation. In order to prevent airway disease and other health problem, it is advisable that people should reduce the exposure time when they worship at the temple with heavy incense smokes, and ventilate their house when they burn incense at home.

RCSN-3 cells are a cloned cell line derived from the substantia nigra of an adult rat. The cell line grows in monolayer and does not require differentiation to express catecholaminergic traits, such as (i) tyrosine hydroxylase; (ii) dopamine release; (iii) dopamine transport; (iv) norepinephrine transport; (v) monoamine oxidase (MAO)-A expression, but not MAO-B; (vi) formation of neuromelanin; (vii) vesicular monoamine transporter-2 (VMAT-2) expression. In addition, this cell line expresses serotonin transporters, divalent metal transporter, DMT1, dopamine receptor 1 mRNA under proliferating conditions, and dopamine receptor 5 mRNA after incubation with dopamine or dicoumarol. Expression of dopamine receptors D2, D3 and D4 mRNA were not detected in proliferating cells or when the cells were treated with dopamine, CuSO4, dicoumarol or dopamine-copper complex. Angiotensin II receptor mRNA was also found to be expressed, but it underwent down regulation in the presence of aminochrome. Total quinone reductase activity corresponded 94% to DT-diaphorase. The cells also express antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase. This cell line is a suitablein vitro model for studies of dopamine metabolism, since under proliferating conditions the cells express all the pertinent markers.

Human mast cells are multifunctional cells capable of a wide variety of inflammatory responses. Berberine (BER), baicalein (BAI), and triptolide (TRI) are compounds isolated from traditional Chinese medicinal herbs and have been shown to have anti-inflammatory effects. We examined their effects on the production of inflammatory cytokines from IL-1β-activated human mast cells (HMC-1). Two ml of HMC-1 at 1 x 106 cells / mL were cultured with BER, BAI, and TRI (at predetermined non-toxic concentrations) in the presence or absence of IL-1β (10 ng / mL) for 24 hrs. The supernatants were harvested and assayed for IL-6, IL-8, and MCP-1 by ELISA. The compounds alone did not induce cytokine production from HMC-1. However, BAI (15 and 30 μM) significantly inhibited the production of IL-6, IL-8 and MCP-1 (all p < 0.005) from IL-1β-activated HMC-1 in a dose dependent fashion. TRI (10 and 20 nM) significantly inhibited IL-8 and MCP-1 (p < 0.001 and < 0.05, respectively), but not IL-6 production from IL-1β-activated HMC-1. BER (25 and 50 μM) inhibited the production of MCP-1 (p = 0.02 and = 0.005, respectively), had no effect on IL-6 production, and enhanced the production of IL-8 at 25 μM (p = 0.01) from IL-1β-activated HMC-1. The results show that BER, BAI, and TRI had differential effects on the production of inflammatory cytokines. Since BAI has the most potent inhibitory effect, it may lead to new anti-inflammatory therapy. (Supported by the Chair of Excellence in Medicine, the Ruth R. Harris Endowment, and RDC of ETSU)