• Toxicological Research
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• v.33 no.4
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• pp.283-290
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• 2017

The host immune system is the first line of host defense, consisting mainly of innate and adaptive immunity. Immunity must be maintained, orchestrated, and harmonized, since overactivation of immune responses can lead to inflammation and autoimmune diseases, while immune deficiency can lead to infectious diseases. We investigated the regulation of innate and adaptive immune cell activation by Artemisia capillaris and its components (ursolic acid, hyperoside, scopoletin, and scopolin). Macrophage phagocytic activity was determined using fluorescently labeled Escherichia coli, as an indicator of innate immune activation. Concanavalin A (ConA)- and lipopolysaccharide (LPS)-induced splenocyte proliferation was analyzed as surrogate markers for cellular and humoral adaptive immunity, respectively. Neither A. capillaris water extract (WAC) nor ethanol extract (EAC) greatly inhibited macrophage phagocytic activity. In contrast, WAC suppressed ConA- and LPS-induced proliferation of primary mouse splenocytes in a dose-dependent manner. Similarly, EAC inhibited ConA- and LPS-induced splenocyte proliferation. Oral administration of WAC in mice decreased ConA- and LPS-induced splenocyte proliferation, while that of EAC suppressed LPS-induced splenocyte proliferation. Repeated administration of WAC in mice inhibited ConA- and LPS-induced splenocyte proliferation. Ursolic acid, scopoletin, and scopolin reduced ConA- and LPS-induced primary mouse splenocyte proliferation, while hyperoside did not show such activity. These results indicate that A. capillaris and its components, ursolic acid, scopoletin, and scopolin, suppress ConA- and LPS-induced adaptive immune cell activation. The results suggest that A. capillaris is useful as a regulator of adaptive immunity for diseases involving excessive immune response activation.

• Biomolecules & Therapeutics
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• v.2 no.3
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• pp.236-243
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• 1994

A splenocyte culture system supplemented with liver microsomes was developed to detect immunotoxic chemicals which require metabolic activation using cyclophosphamide as a positive standard. When liver microsomes were added to splenocyte cultures isolated from female B6C3Fl mice, the proliferation of splenocytes by lipopolysaccharide (LPS) was increased and the proliferation by concanavalin A (Con A) was decreased. However, when compared with each corresponding control, cyclophophamide was successfully activated to metabolites capable of suppressing Iymphoproliferative responses. This suppression was clearly dependent upon the amounts of microsomes added and/or the concentration of cyclophosphamide exposed. In these cultures, the proliferation of splenocytes was suppressed when the cells were exposed to cyclophosphamide on the day of culture initiation. On the other hand, microsome was responsible for the increase in LPS mitogenicity and NADPH was responsible for the decrease in Con A mitogenicity. Finally, our present culture system was compared with the hepatocyte-splenocyte coculture system which we had developed earlier. We found that the hepatocyte-splenocyte coculture was better able to activate cyclophosphamide to metabolites capable of suppressing the antibody response to sheep erythrocytes. Although our present culture system was relatively poor to activate cyclophosphamide in cultures for antibody response, it will be useful as a simple screening method to detect suppression of certain in vitro immunotoxic parameters like LPS mitogenicity by chemicals which require metabolism.

• The Korean Journal of Food And Nutrition
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• v.24 no.1
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• pp.65-70
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• 2011

This study was performed to investigate the in vitro effect of a corn water extract on immune function. Splenocyte proliferation was determined by the MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl terazolium bromide) assay after preparing asingle cell suspension. Production of macrophage-secreted interleukin(IL)-$1{\beta}$, IL-6, and interferon(IFN)-${\gamma}$, was detected by ELISA using a cytokine assay kit. After a 48-hr incubation with mitogens(ConA or lipopolysaccharide), mice splenocyte proliferation increased with the addition of a corn water extract supplement at 10, 50, 100, 250, 500, or $1,000\;{\mu}g/m\ell$. Production of IL-$1{\beta}$, IL-6, and IFN-${\gamma}$ increased in treatments supplemented with the corn water extract. In an in vitro study, splenocyte proliferation increased when $50\sim1,000\;{\mu}\ell/m\ell$ corn water extract was added. In an ex vivo experiment, the highest production of cytokines by activated peritoneal macrophages was observed in mice orally administered 500 mg/kg body weight/day.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.6
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• pp.954-961
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• 2007

The experiments were conducted to determine effects of a mixture of conjugated linoleic acid (CLA) isomers on T cell subpopulations and responsiveness to mitogen of splenocytes in male broiler chicks. In experiment 1, birds (8-d old) were fed basal, CLA-(CLA) and safflower oil-supplemented (SA) diets which were formulated by supplementary 10 g CLA or safflower oil/kg to the basal diet for 14 d. Broiler starter diet, which mainly consisted of corn and soybean meal, was served as the basal diet. Proliferative response and interleukin (IL)-2-like activity stimulated by concanavalin (Con) A at a concentration of $10{\mu}g/ml$ of splenocytes in chicks fed the CLA diet were greater than in chicks fed the SA diet, but not at $20{\mu}g$ Con A/ml. Percentage of CD3-positive T cells in splenocytes did not differ between chicks fed the SA diet and CLA. Ratio of CD4-positive T cells to CD8- positive T cells was significantly affected by dietary fat source. In experiment 2, broiler chicks (1-d old) were fed the same diets as in experiment 1 for 14 d. Results of splenocyte proliferation to Con A were similar to those in experiment 1, but phytohemaggulutinin (PHA)- or pokeweed mitogen (PWM)- induced splenocyte proliferation did not differ between the CLA and SA fed groups. Supplementation with SA or CLA to the basal diet tended to have a depressive effect on the proliferation, with the greater effect being that of SA. In experiment 3, effect of an addition of CLA to splenocyte culture medium on splenocyte proliferation was determined. An addition of CLA to the culture medium resulted in reduction of the splenocyte proliferation to Con A, but an addition of linoleic acid. When PWM and PHA were used as mitogen, the inhibitory effect of CLA and linoleic acid on the proliferation did not differ. The results suggested that the effect of dietary CLA on splenocyte proliferation was similar to that of SA, although the effect of dietary CLA on sub-populations was slightly different from that of dietary SA. Further studies are needed to clarify whether use of CLA would be beneficial for maintaining or enhancing T cell immunity in chicks.

• Journal of Life Science
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• v.21 no.8
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• pp.1176-1182
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• 2011

The present research was conducted to investigate the effect of moderate exercise training on splenocyte inflammatory cytokines production in high fat diet-induced obese mice. To determine the effects of exercise training and low fat diet on splenocyte, C57BL/6 male mice (4 weeks aged, n=20) were fed a high fat diet (45% fat diet) for 5 weeks so that obesity was achieved intentionally. These obese mice were then divided into 2 groups; HLC (low fat diet and control n=10), and HLE (low fat diet and exercise n=10). HLE mice performed 8 weeks of exercise training on a motorized treadmill by running for 30-60 min/day at 10-22 m/min, 0% grade, five times per week. After exercise training, all the splenocyte was collected and Con A (concanvalin A, 10${\mu}g/ml$) was used to stimulate the cells, after which IL-1${\beta}$, IL-6, TNF-${\alpha}$ were measured by bio plex. Independent t-test was used and p value under 0.05 was considered a statistical significance. Splenocyte IL-1${\beta}$, IL-6, TNF-${\alpha}$ production of HLE stimulated by Con A was significantly lower than that of HLC (p<0.01). These findings suggest that moderate exercise has beneficial effects on splenocyte inflammatory cytokines in high fat diet induced obese mice.

• The Korean Journal of Food And Nutrition
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• v.21 no.3
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• pp.269-274
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• 2008

Plants have long been used as a food source in Korea. In this study, we investigated the combined immunomodulative effects of a water extract mixture of(pine needles, Sedum sarmentosum Bunge, hijkiaorme, buckwheat and Peril a leaves) on Balb/c mice $7{\sim}8$ weeks old. The mice were fed a chow diet ad libitum and the plant extract was orally administered every other day for four weeks at two different concentrations(50 and 500 mg/kg BW). After preparing the single-cell suspension, splenocyte proliferation was determined by the MTT(3-[4,5-di-methylthiazol-2-y]-2,5-diphenyl terazolium bromide) assay. After 48hrs of incubation with the mitogens(ConA or LPS) splenocyte from the mice groups administered 50 and 500 mg/kg BW of the plant extract showed a significant increased in proliferation compared to the control group. A hemolytic plague forming cell assay was used to indicate antibody production against sheep red blood cells(SRBC). The number of antibody-secreting cells T-dependent antigen. The result of this study suggest that supplementation with this plant extract may regulate immune function by increasing splenocyte proliferation and the number of plaque forming cells.

• Journal of the Korean Society of Food Culture
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• v.35 no.2
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• pp.215-223
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• 2020

Ganjang and doenjang are known as major fermented soy-based foods in Koreans. Current investigations have proved that fermented soybean foods impart anti-cancer, anti-obesity, and anti-diabetic effects. The aim of this study was to evaluate the anti-cancer activities of commercialized soy food, Ganjang, as a function of aging period. The test groups were classified into four time periods-short (under 5 years, S group), mid (under 10 years, M group), long (under 15 years, L group), and eternal (over 15 years, E group). The anti-cancer effects of Ganjang were determined by cell cytotoxicity assay of three types of cancer cell lines and splenocyte proliferation assay. Besides these assays, we also analyzed NK cell activity for cancer immunotherapy. The results show that the anti-cancer effect increased in the S and M period aging groups for all three cancer cell lines. Interestingly, similar to the anti-cancer result, splenocyte proliferation and NK activity showed the highest effect in the S and M groups. In contrast, Japanese ganjang-treated (JG1, JG2) groups and E group showed significantly reduced splenocyte proliferation. Collectively, these results suggest that the short and middle periods of traditional fermented Ganjang might have potential anti-cancer activities.

• Journal of Nutrition and Health
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• v.31 no.6
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• pp.973-980
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• 1998

To investigate the effect of sea tangle on immune function in normal and diabetic states, 10-week old ICR mice were feed control(C) and sea tangle(5) diets containing 5%(w/w) cellulose and 13.6%(w/w) dry sea tangle for 4 weeks. After 4 weeks, three quarters of mice(CD and SD) were made diabetic by intramuscular injection of streptozotocin(150mg/kg bw). On the 4th day after diabetes was apparent by urinary glucose, one third of diabetic mire(CDG and SDG) were treated with glipizide(20mg/kg bw) and the other third(CDM and SDM) with metformin (500mg/kg bw) orally. Spleen weights of diabetic mice with no hypoglycemic drug treatment appeared to be higher in the sea tangle group(SD) than in control(CD), but were not different when drugs were administered. Data on splenocyte proliferation stimulated by lipopolysaccaride from Salmonella abortus equi(0.l$\mu\textrm{g}$/ml) showed that sea tangle increased mitogen response in normal mice(C group vs S group) and appeared to have the same effect in diabetic mice with or without drug treatment. Splenocyte proliferation induced by concanavalin A(0.1$\mu\textrm{g}$/ml) also showed similar results, although there were not statistically significant. Concentration of interleukin-2(IL-2) released from splenocytes of the S group seemed higher than from the C group, but the IL-2 concentrations were not different among six diabetic groups. Results of fatty acid compositions of splenocyte phospholipids showed that diabetes reduced arachidonic acid/linoleic acid ratios and that sea tangle intake and glipizide treatments increased contents of polyunsaturated fatty acids. It is concluded that dietary sea tangle has a positive effect on splenocyte proliferation under normal condition and could have the same effect under diabetic conditions. IL-2 appears to be one of factors mediating the effect but involvement of membrane fatty arid changes and other unknown factors needs lurker Investigation. (Korean J Nutrition 31(6) : 973-980, 1998)

• The Journal of Internal Korean Medicine
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• v.30 no.1
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• pp.9-23
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• 2009

Objectives : Asthma is a chronic inflammatory disorder of the airways by many cells such as mast cells, Th2 lymphocytes and eosinophile. The present study was aimed to evaluate the effects of Salviae miltiorrhizae (SM) on T cell cytokine production, mast cells. and eosinophils, Methods : We screened 13 herbs to find compounds with potential to control Th cytokine production. using concanavalin A (con A)-activated splenocyte cultures. Con A-activated $IFN-\gamma$ and IL-4 levels in supernatants of splenocyte cultures. Bone marrow derived mast cells (BMMC) were incubated with SM and then the expressions of membrane proteins of BMMC were analyzed by fluorescence activated cell sorter (FACS). BALB/c mice sensitized to ovalbumin (OVA) were challenged with aerosolized OVA for 6 weeks. During the last weeks some mice were treated with SM. Then eosinophils in bronchoalveolar lavage fluid (BALf) were counted and pathologic changes of lung tissue were observed with hematoxylin-eosin stain. Results : SM increased $IFN-\gamma$ level on splenocyte culture significantly. but had no significant effects on expressions of ICAM-1, CD62L, integrin $a_4$. c-kit, IL-3 receptors. CD11a, or IgE receptors of BMMC. SM treatment significantly inhibited eosinophil infiltrates in BALf and peribronchial lung inflammation. Conculusions : The present data suggested that SM may have an effect on Th cytokine secretion and eosinophils associated with asthma responses. Therefore SM might be of therapeutic value in treating asthma.

• Food Science of Animal Resources
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• v.26 no.1
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• pp.113-120
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• 2006

Insulin-like growth factor-I(IGF-I) rich fraction, which was obtained molecules ranged between 30 and 1 kDa, was fractionated by ultrafiltration from bovine colostral whey. IGF-I rich fraction was confirmed by SDS-PACE and western blotting and then the quantity of IGF-I was measured by sandwich ELISA. ICF-I concentration in IGF-I rich fraction was 10 ng/mg proteins. IGF-I rich fraction, standard IGF-I and colostral whey weie treated to murine peritoneal macrophages. And then we experimented that effect of immune activity on macrophage and splenocyte. As a result, in group treated with IGF-I rich fraction $1{mu}g/mL$, production of interleukin-6 and nitric oxide were 9.85 ng and $17.17{\mu}M$ and production of phagocytosis, tumor necrosis factor-${\alpha}\;and\;H_{2}O_{2}$ were 78.3, 34.5 and 6% compared to the control group. In splenocyte immune response, B cell and T cell proliferation and NK cell activity were 103, 126 and 22.2% in group treated with IGF-I rich fraction $1{\mu}g/mL$ to compared to the control, respectively.