• Korean Journal of Poultry Science
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• v.34 no.1
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• pp.31-36
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• 2007

This experiment was conducted to determine the effect of betaine on immune response in laying hens. A total of 72 ISA-brown laying hens were divided into four groups of 18 hens each and fed corn-soybean meal based diets with addition of 0, 300, 600 and 1,200 ppm betaine for four weeks. The effect of betaine on splenocyte proliferations with mitogens, concanavalin A(Con A) and pokeweed mitogen(PWM), were assayed after incubation using [3H] thymidine uptake. Proliferations of splenocyte were significantly increased by activation of mitogen Con A or PWM. Mitogen effects of Con A were increased by Con A plus betaine injection(0.1 mM), whereas PWM effects did not affect in PWM plus betaine injection(0.1 mM) in vitro. Splenocyte of laying hens fed betaine tended to proliferate in the presence of PWM, but appeared to be slightly suppressed in the presence of Con A in vivo. Proliferation of splenocytes which were stimulated by Con A or Con A+betaine injection(0.1 mM) were increased in dietary 600 ppm betaine, but inhibited in dietary 1,200 ppm betaine supplementation. Spleen weights and sheep red blood cell(SRBC) titers of hens fed betaine tended to increase compared to those of control, but were not significantly different. These results suggested that betaine could increase splenocyte proliferation in vitro.

• Parasites, Hosts and Diseases
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• v.33 no.3
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• pp.195-200
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• 1995

The presence of biological response modifiers (BRMI-like effect was confirmed in peritoneal exudate (PE) of ToxopLnsmo gondii-infected ICR mice which inhibited Concanavalin A (Con A)-induced peritoneal Iymphocyte (PL) proliferation. During 5 days of PL incubation with $10{\;}\mu\textrm{g}/ml$ Con A with or without PE, 3H-thymidine uptake was measured for the last 24 hrs. Compared to uninduced control, PL proliferated by 7.3-fold with Con A induction_ When PE of infected mice was added, PL proliferation was inhibited by $74.0{\;}{\pm}{\;}11.9%$ whereas inhibition by PE of normal mice was $16.4{\;}{\pm}8.3%$. Inhibitory effect of PE increased exponentially from 3 days up to 4-5 days of survival after the infection. Inhibitory activity of PE was decreased concentration-dependently. Also the inhibition was diminished when the PE was treated with heat of $95^{\circ}C$ for 10 min orprecipitated with 10% trichloroacetic acid (TCA). In SDS-PAGE of PE, many minor bands appeared newly. Heat-labile protein molecule in PE exerted inhibitory activity to Con A- induced Iymphocyte proliferation.

• The Korean Journal of Pharmacology
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• v.27 no.2
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• pp.135-144
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• 1991

Using T-lymphocytes obtained from rat peripheral blood, we found that the 44kD/pI6.8 protein was the major phosphoprotein of T-lymphocytes under basal condition, and that the 44kD/pI6.3 protein was a new phosphoprotein appeared in T-lymphocytes stimulated with ${\beta}-agonist$. The phosphorylation of the 44kD/pI6.3 protein was also induced by forskolin but inhibited by H-8 pretreatment. To clarify the character of the 44kD/pI6.3 protein, we used Con-A and kinase inhibitors, H-7 and W-7. Con-A stimulation induced phosphorylation of 44kD/pI 6.3 protein but that was inhibited by W-7 pretreatment. The phosphorytation of 44kD/pI6.3 protein was not induced by the PKC activator, PMA. Instead, the phosphorylation of 44kD/pI6.8 protein was reduced by H-7, a PKC inhibitor. From the above results,it can be concluded that the 44kD/pI6.3 protein can be a common substrate for A-kinase and CaM kinase. The two dimensional tryptic peptide mapping revealed that the 44kD/pI6.8 and 44kD/pI6.3 proteins are different.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.2
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• pp.174-180
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• 2016

In this study, the effects of fermented goat milk (F-GM) on immunological activity and physical strength were examined. Splenocytes obtained from mice administered with F-GM showed increased responsinveness to mitogens, concanavalin-A (ConA), and lipopolysaccharide (LPS). Treatment with F-GM also significantly augmented production of interleukin (IL)-2 and interferon (IFN)-${\gamma}$, but not IL-4 or IL-10 from ConA-stimulated splenocytes. The activity of F-GM administration to enhance production of IL-2 and IFN-${\gamma}$ was confirmed based on mRNA expression of these cytokines by reverse transcription polymerase chain reaction. After immunization with keyhole limpet hemocyanin (KLH, 20 mg/mouse), mice administered F-GM showed significantly higher antibody titers against KLH than those of phosphate-buffered saline-treated mice, and showed the highest titer 5 weeks after KLH immunization. Analysis for determining isotypes of antibodies revealed that administration of F-GM elicited KLH-specific antibody titers of IgG1, IgG2a, and IgM. In a delayed type hypersensitivity (DTH) test carried out 7 weeks after the primary immunization, F-GM-treated mice showed a higher DTH reaction than the control mice. Furthermore, the swimming test found that administration of F-GM significantly increased swimming time. These results suggest that administration of F-GM enhances not only immune responses against antigens but also physical strength.

• IMMUNE NETWORK
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• v.1 no.3
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• pp.213-220
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• 2001

Background: A previous study has shown that Euonymus alatus (EA) has an antidotic activities against inflammation, suggesting possibility that EA can exert this beneficial effects to liver injury by an initial protection against drug-induced hepatocyte demage. The present study was undertaken to evaluate the protective effect of EA-extract on experimentally induced hepatitis in ICR mice and to investigate some mechanisms responsible for its action. Methods: Water EA extract was used in this experiments. The mice received i.p. a dose of 700 mg/kg galactosamine (GalN) together with $5{\mu}g/kg$ of endotoxin (LPS), or received i.v. 12 mg/kg of concanavalin A (Con A). EA (4 mg/mouse) was administrated on day -2, -1 and 0 before induction of liver injury. Liver injury was assessed by measurement of serum alanin amino-transferase (SGPT) levels on 9 hr after GaIN.LPS, or 8 hr after con A administration. Results: Treatment with either GaIN or LPS alone did not cause hepatitis. However, simultaneous administration of GalN and LPS to mice resulted in LPS-dose dependent fulminant hepatitis. GaLN/LPS-induced liver injury was reduced when mice were given EA for 3 days before induction. This preventive effect of Ea was more prominent when EA was given by intraperitoneal route rather then by oral route. Pretreatment of EA or dexamethasone inhibited significantly $TNF{\alpha}$ production in GalL/LPS-injured mice. However, EA-treatment did not influence $TNF{\alpha}$-induced hepatitis in GalN-sensitized mice, suggesting that $TNF{\alpha}$ is likely to act as one of final mediators of endotoxin action and the protective effect of EA might be manifested chiefly by inhibition of endotoxin-induced $TNF{\alpha}$ production, not by blocking the $TNF{\alpha}$-action. Injection of Con A into mice evoked remarkable liver injury in a dose dependent fashion. This liver damage was reduced by EA-pretreatment. Dexamethasone significantly reduced both GalL/LPS-induced and Con A-induced liver damages, showing synergism with EA. However, indomethacin reduced only GalN/ LPS-induced hepatitis, not for Con A-induced hepatitis. Conclusion: These results led to the conclusion that EA may be able to contribute at least in part to prevent the drug-induced hepatotoxicity, and that its anti-hepatitis effects might be manifested directly by modulation of endogenous mediators, such as leukotriese D4, $TNF{\alpha}$ and free radical, and indirectly by regulation of immune mediated responses. Also these results suggested that EA could be developed as a potential antidotic agent.

• The Korean Journal of Pharmacology
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• v.29 no.1
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• pp.121-130
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• 1993

This study was done to classify the proteins involved in the specific phosphorylation using the rat peripheral blood lymphocytes (rPBL) stimulated with mitogens, phorbol 12-myristate 13-acetate (PMA) and concanavalin A (Con A). The lymphocytes were incubated with $^{32}P-orthophosphate$ before PMA or Con A stimulation. The migration patterns of the phosphorylated proteins of mitogen-treated rPBL in two dimensional electrophoretic fields were analyzed after autoradiography. The stimulation of the lymphocytes with PMA and Con A increased the phosphorylation of thirteen protein fractions. The phosphorylation intensities of the protein spots differ to the treatments of the cells with specific kinase inhibitors, H-7 and W-7. These protein fractions were grouped into 3 classes, namely, PKC-mediated, CaM kinase-mediated, and other kinase mediated proteins. The effect of the duration of the stimulation on the phosphorylated behaviors occurred concurrently, not sequentially, although each individual protein fraction had a different time for the peak phosphorylation during the stimulation period upto 30 minutes. The phosphoproteins found in the cytosolic soluble fraction were phosphorylated prior to those in the pellet, whose phosphorylations were sustained at a high level for over 10 minutes. The above results suggest that the early events in lymphocyte activation involve 3 different sets of proteins which are phosphorylated by CaM kinase, PKC and other kinases, and those kinases do not work sequentially, but rather, independently or cooperatively.

• Journal of fish pathology
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• v.20 no.2
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• pp.189-200
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• 2007

To investigate the innate immune response involved in early stage of anti-viral defence, carps were injected with UV-inactivated spring viraemia of carp virus (SVCV), poly inosinic:cytidylic acid (Poly I:C) and concanavalin A (Con A), respectively and examined lysozyme activity, serum complement activity and chemiluminescent (CL) response of leucocytes isolated from head kidney at 3 days post-injection. There was no significant difference in plasma lysozyme activities among all experimental groups. However, lysozyme activities of head kidney in the groups injected with antiviral activity inducers were significantly higher than those of the control injected with physiological saline. Bactericidal activities of serum of the groups injected with antiviral activity inducers were not significantly different from control group. However, the CL responses were significantly higher at lower dose of Poly I:C and Con A, whilst dose-dependent increase was shown in UV-inactivated SVCV-injected group. In the challenge test with 1×104 TCID50/fish of SVCV at 4 days post-injection, UV-inactivated SVCV- and Poly I:C-injected groups showed higher relative percent survival (RPS) than Con A-injected group. Furthermore, strong protection was observed in the group injected higher dose of Poly I:C although showed lower activities in lysozyme and CL response. These results suggested that Poly I:C might stimulate other factors belonging to non-specific immune system have induced protective immunity against the SVCV challenged.

• 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.

• Toxicological Research
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• v.34 no.1
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• pp.65-73
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• 2018

Pretreatment of low-dose lipopolysaccharide (LPS) induces a hyporesponsive state to subsequent secondary challenge with high-dose LPS in innate immune cells, whereas super-low-dose LPS results in augmented expression of pro-inflammatory cytokines. However, little is known about the difference between super-low-dose and low-dose LPS pretreatments on immune cell-mediated inflammatory and hepatic acute-phase responses to secondary LPS. In the present study, RAW 264.7 cells, EL4 cells, and Hepa-1c1c7 cells were pretreated with super-low-dose LPS (SL-LPS: 50 pg/mL) or low-dose LPS (L-LPS: 50 ng/mL) in fresh complete medium once a day for 2~3 days and then cultured in fresh complete medium for 24 hr or 48 hr in the presence or absence of LPS ($1{\sim}10{\mu}g/mL$) or concanavalin A (Con A). SL-LPS pretreatment strongly enhanced the LPS-induced production of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, TNF-${\alpha}$/IL-10, prostaglandin E2 ($PGE_2$), and nitric oxide (NO) by RAW 264.7 cells compared to the control, whereas L-LPS increased IL-6 and NO production only. SL-LPS strongly augmented the Con A-induced ratios of interferon (IFN)-${\gamma}$/IL-10 in EL4 cells but decreased the LPS-induced ratios of IFN-${\gamma}$/IL-10 compared to the control, while L-LPS decreased the Con A- and LPS-induced ratios of IFN-${\gamma}$/IL-10. SL-LPS enhanced the LPS-induced production of IL-6 by Hepa1c1c-7 cells compared to the control, while L-LPS increased IL-6 but decreased IL-$1{\beta}$ and C reactive protein (CRP) levels. SL-LPS pretreatment strongly enhanced the LPS-induced production of TNF-${\alpha}$, IL-6, IL-10, $PGE_2$, and NO in RAW 264.7 cells, and the IL-6, IL-$1{\beta}$, and CRP levels in Hepa1c1c-7 cells, as well as the ratios of IFN-${\gamma}$/IL-10 in LPS- and Con A-stimulated EL4 cells compared to L-LPS. These findings suggest that pre-conditioning of SL-LPS may contribute to the mortality to secondary infection in sepsis rather than pre-conditioning of L-LPS.

• 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.