Splenocyte-mediated immune enhancing activity of Sargassum horneri extracts / 한국영양학회지

PURPOSE: This study examined the immunological activity and optimized the mixture conditions of Sargassum horneri (S. horneri) extracts in vitro and in vivo models.METHODS: S. horneri was extracted using three different methods: hot water extraction (HWE), 50% ethanol extraction (EE), and supercritical fluid extraction (SFE). Splenocyte proliferation and cytokine production (Interleukin-2 and Interferon-γ) were measured using a WST-1 assay and enzyme-linked immunosorbent assay, respectively. The levels of nitric oxide and T cell activation production were measured using a Griess assay and flow cytometry, respectively. The natural killer (NK) cell activity was determined using an EZ-LDH kit.RESULTS: Among the three different types of extracts, HWE showed the highest levels of splenocyte proliferation and cytokine production in vitro. In the animal model, three different types of extracts were administrated for 14 days (once/day) at 50 and 100 mg/kg body weight. HWE and SFE showed a high level of splenocyte proliferation and cytokine production in the with and without mitogen-treated groups, whereas EE administration did not induce the splenocyte activation. When RAW264.7 macrophage cells were treated with different mixtures (HWE with 5, 10, 15, 20% of SFE) to determine the optimal mixture ratio of HWE and SFE, the levels of nitric oxide and cytokine production increased strongly in the HWE with 5% and 10% of SFE containing group. In the animal model, HWE with 5% and 10% of SFE mixture administration increased the levels of splenocyte proliferation, cytokine production, and activated CD4⁺ cell population significantly, with the highest level observed in the HWE with 5% of SFE group. Moreover, the NK cell activity was increased significantly in the HWE with 5% of SFE mixture-treated group compared to the control group.CONCLUSION: The optimal mixture condition of S. horneri with immune-enhancing activity is the HWE with 5% of SFE mixture. These results confirmed that the extracts of S. horneri and its mixtures are potential candidate materials for immune enhancement.

Immune Efficacy and Safety of Fucoidan Extracted from Gagome Kombu <i>(Kjellmaniella crassifolia)</i> in Healthy Japanese Subjects / 日本補完代替医療学会誌

Objective: We previously reported the immune-enhancing behavior of fucoidan, a sulfated polysaccharide extracted from Gagome kombu (GKF), both in vitro and in animal studies. In the present study, we evaluated the immune efficacy and safety of GKF in healthy Japanese adults.Methods: In this randomized, double blind, placebo-controlled study, 30 subjects who ingested GKF (200 mg/day) or placebo for 4 weeks were enrolled. For evaluation of efficacy, phytohemagglutinin-stimulated cytokine production in whole blood cells was measured. For evaluation of safety, blood chemistry analysis, hematological analysis, and urinalysis were conducted.Results: Almost all cytokine production decreased in samples from the placebo group during the test period. Ingestion of GKF for 4 weeks significantly suppressed the decrease in production of the T helper 1 (Th1)-type cytokines interferon-γ and interleukin-12 as well as the Th1:Th2 ratio. There were no adverse clinical changes in blood analysis and urinary analysis, and no serious symptom was observed.Conclusion: These results indicate that GKF is a useful and safe food ingredient to support immune function.

Severe visceral leishmaniasis in children: the relationship between cytokine patterns and clinical features

Introduction The relationship between severe clinical manifestations of visceral leishmaniasis (VL) and immune response profiles has not yet been clarified, despite numerous studies on the subject. This study aimed to investigate the relationship between cytokine profiles and the presence of immunological markers associated with clinical manifestations and, particularly, signs of severity, as defined in a protocol drafted by the Ministry of Health (Brazil). Methods We conducted a prospective, descriptive study between May 2008 and December 2009. This study was based on an assessment of all pediatric patients with VL who were observed in a reference hospital in Maranhão. Results Among 27 children, 55.5% presented with more than one sign of severity or warning sign. Patients without signs of severity or warning signs and patients with only one warning sign had the highest interferon-gamma (IFN-γ) levels, although their interleukin 10 (IL-10) levels were also elevated. In contrast, patients with the features of severe disease had the lowest IFN-γ levels. Three patients who presented with more than two signs of severe disease died; these patients had undetectable interleukin 2 (IL-2) and IFN-γ levels and low IL-10 levels, which varied between 0 and 36.8pg/mL. Conclusions Our results showed that disease severity was associated with low IFN-γ levels and elevated IL-10 levels. However, further studies with larger samples are needed to better characterize the relationship between disease severity and cytokine levels, with the aim of identifying immunological markers of active-disease severity. .

Sports and Immunity / 全日本鍼灸学会雑誌

In this review the author scrutinizes the effects of exercise on the immune system on two points. One is the effect of usual proper exercise on the immune system, and the other is the acute effect of intense exercise on the immune system. Concerning the first point, exercise augments physiological activities such as circulation, metabolism and physiological regulatory systems. These effects cause augmentation of the defense mechanisms, i.e. fortification of barrier systems, increases in the number of cells in the immune system and augmentation of cell migration and potentiation of cytokine production. As a result, daily exercise habits show prophylactic effects against infection, improvement in predisposition and depressed immunosuppression by senescence. Concerning the second point, acute intense exercise triggers activation of the hypothalamus-pituitary-adrenal (HPA) axis, and, as a result, the immune response is suppressed by corticosteroid. Moreover, intense exercise induces harmful factors which suppress the immune system. Proper daily exercise is, therefore, recommended to maintain the immune system in good condition.

Inflammatory Mediators Modulate NK Cell-stimulating Activity of Dendritic Cells by Inducing Development of Polarized Effector Function

BACKGROUND: It is well established that cross talk between natural killer (NK) cells and myeloid dendritic cells (DC) leads to NK cell activation and DC maturation. In the present study, we investigated whether type 1-polarized DC (DC1) matured in the presence of IFN-gamma and type 2-polarized DC (DC2) matured in the presence of PGE2 can differentially activate NK cells. METHODS: In order to generate DC, plastic adherent monocytes were cultured in RPMI 1640 containing GM-CSF and IL-4. At day 6, maturation was induced by culturing the cells for 2 days with cytokines or PGE2 in the presence or absence of LPS. Each population of DC was cocultured with NK cells for 24 h. The antigen expression on DC was analyzed by flow cytometry and cytokine production in culture supernatant was measured by ELISA or a bioassay for TNF-alpha determination. NK cell-mediated lysis was determined using a standard 4 h chromium release assay. RESULTS: DC2, unlike DC1, had weak, if any, ability to induce NK cell activation as measured by IFN-gamma production and cytolytic activity. DC2 were weakly stimulated by activated NK cells compared to DC1. In addition, IFN-gamma-primed mature DC appeared to be most resistant to active NK cell-mediated lysis even at a high NK cell/DC ratio. On the other hand, PGE2-primed DC were less resistant to feedback regulation by NK cells than IFN-gamma-primed mature DC. Finally, we showed that the differential effect of two types of DC population on NK cell activity is not due to differences in their ability to form conjugates with NK cells. CONCLUSION: These results suggest that different combinations of inflammatory mediators differentially affect the effector function of DC and, as a result, the function of NK cells, eventually leading to distinct levels of activation in adaptive immunity.

Age-dependent changes of B cell and T cell-mediated immune responses in naive C3H/HeJ mice under regular mouse chow feeding conditions / 천식및알레르기

BACKGROUND AND OBJECTIVE: Murine system for studying allergic diseases has been popular in the fields of food allergy and development of their therapeutic strategies. However, there has been no information about the age-dependent changes of natural immune responses of naive C3H/HeJ mice. The purpose of this study was to evaluate the age-dependent changes of B and T-cell mediated immunologic parameters in naive C3H/HeJ mice, which can provide information for experimental planning and analysis of research results. SUBJECTS AND METHODS: Eight naive, female, 5-week-old C3H/HeJ mice were grown under the regular mouse chow feeding conditions for 6 weeks. Sera were obtained at week (w) 5, w6, w8 and w10 for measuring total and chow-specific IgE, IgG1 and IgG2a antibodies. Splenocyte proliferation (at w8 and w10) and cytokine production (at w6, w8 and w10) were evaluated with or without Con A stimulation with pooled splenocytes from two mice of each age group. Serum antibodies and cytokines (IL-4, IL-5, IL-12, INF-gamma, TGF-beta1) were measured by ELISA. Using RT-PCR, IL-4 and INF-gamma mRNA expressions were measured in Peyer's patch and spleen tissue at w10. RESULTS: The levels of total IgE and IgG1 were increased by age while the level of IgG2a was decreased. Chow-specific IgE and IgG2a responses were neglectable through out the whole experimental period (20-30 ng/ml or less). Chow-specific IgG1 levels were measured in the significant concentrations (200-300 ng/ml) but there was no age-dependent change through out the experiment. Con A stimulated-splenocyte proliferation indexes were variable according to the culture-durations and ages of mice. The higher proliferation indexes were observed in the wells receiving thymidine pulse at 48-hour culture, especially in the mice at w10. Con A stimulated IL-4 production in the 72-hour splenocyte culture supernatant was significantly increased at w8, and w10 while INF-gamma production increased only at w10. The changes in the production of IL-5, IL-12 and TGF-beta did not provide significant information in the present study. The ratio of IL-4/IFN-gamma mRNA expression was higher in Peyer's patch than in the spleen. CONCLUSION: The changes of B-cell and T-cell mediated immunologic parameters were complex and variable according to the age in naive C3H/HeJ mice under regular chow feeding conditions. For that reason, the information from the present study needs to be considered in the course of planning or analysing research/data using murine systems.

Immunologic Alterations of Peripheral Mononuclear Cells by Intravenous Immunoglobulin in the Acute Stage of Kawasaki disease

PURPOSE: High dose intravenous immunoglobulin (IVIG) therapy is effective in reducing the incidence of coronary artery aneurysm in Kawasaki disease (KD) patients, however, the precise mechanisms by which IVIG reverses the immune activation is unknown. METHODS: The sera and peripheral mononuclear cells (PBMCs) were obtained from 10 KD patients in the acute stage (24 hrs before) and 24 hrs after one dose of IVIG (2g/kg) treatment. We measured serum interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) and compared them to the messenger RNA expression of the PBMCs by reverse transcription polymerase chain reaction. We also investigated the cytokines' or immunoglobulins' production of PBMCs which were stimulated with phytohemagglutinin-P (PHA-P) or pokeweed mitogen (PWM, without in vitro IVIG treatment) by enzyme-linked immunosorbent assay. RESULTS: The serum levels of IL-6 and TNF-alpha decreased significantly after IVIG treatment. However, the gene expression of IL-6 and TNF-alpha showed no significant difference after IVIG treatment. IL-6 and TNF-alpha productions of PBMCs stimulated with PHA-P decreased significantly after IVIG treatment, however, IL-4 and IFN-gamma production showed no significant differences. IL-6 production of PBMCs stimulated with PWM also decreased significantly after IVIG treatment, but TNF-alpha did not change significantly. The synthesis of IgG, IgM, IgA and IgG1 after IVIG treatment showed a significant decrease, and that of IgG2 showed a slight decrease, but was not statistically significant. CONCLUSION: The results demonstrate that the administration of IVIG in the acute stage of KD resulted in the alterations of activated immunologic abnormalities, especially in monocytes and B cells.