• Microbiology and Biotechnology Letters
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• v.22 no.3
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• pp.240-245
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• 1994

Relation the phage defense mechanism of phage resistant Lactococcus lactis subsp. cremoris ATCC 11602-A1 to its cell wall components was investigated. To determine whether teichoic acid which is known to be one of the phage receptor site present on the cell wall, phage adsorption was examined after treatment 5% TCA(60%$\CIRC $C) and concanavalin A to the cell wall of A1 and parent strain. However, the adsorption rate of two strains did not change. Total amount of phosphate after TCA treatment did not change in both strains, but a difference between the two strains was observed. Ribitol and glycerol, components of teichoic acid, could not be detected in the cell walls of two strains by GC analysis. These results suggest that although teichoic acid was not present in the cell walls of both strains, the composition of cell wall of two strains was not identical. Measurement of amount of protein and SDS-polyacryamide gel electrophoresis were carried out to examine the involvement of cell wall protein in phage resistance, showing that protein is nothing to do with phage adsorption of parent strain, but phage resistance of A1 is related to protein. Cell wall carbohydrates of A1 contained rhamnose, glucose, and galactose. Total amount of carbohydrate of 1% SDS-treated A1 cell wall was reduced to the level of parent strain. The results suggest that phage resistance of A1 was due to the presence of a higher level of carbohydrates then parent strain, and to interaction of carbohydrate and protein.

• Korean Journal of Agricultural Science
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• v.46 no.3
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• pp.579-589
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• 2019

Due to the ban on the use of antibiotics, interest has been increasing for the development of therapeutic agents to treat various diseases using natural resources. Osthole, a natural coumarin compound used in traditional Chinese medicines, exerts an anti-inflammatory effect, but its effects in cows remain unknown. In this study, the effect of osthole on lipopolysaccharide (LPS)- or concanavalin-A (Con-A)- stimulated peripheral blood mononuclear cells (PBMCs) was assessed. Jugular venous blood was collected from Korean calves, and PBMCs were isolated. They were then used to study the immune response of PBMCs to treatment with osthole and LPS or Con-A for 72 h by measuring inflammatory cytokines including tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$) and interferon-${\gamma}$ ($IFN-{\gamma}$). Osthole significantly inhibited the mRNA secretion of $TNF-{\alpha}$ and $IFN-{\gamma}$ in a dose-dependent manner. Therefore, osthole inhibited LPS- or Con-A- induced $TNF-{\alpha}$ and Con-A-induced $IFN-{\gamma}$ production significantly in dose-dependent manner. These results clearly suggest that osthole inhibited the LPS- or Con-A- stimulated upregulation of pro-inflammatory cytokines in a dose-dependent manner, without causing obvious cytotoxic effects. Osthole could also protect cows from LPS- or Con-A- induced endotoxin shock, possibly by inhibiting the production of pro-inflammatory cytokines, which suggests that osthole might be a novel therapeutic agent for the prevention of inflammatory diseases.

• Environmental Analysis Health and Toxicology
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• v.17 no.4
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• pp.325-332
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• 2002

A helper T(Th)1-mediated response is known to enhance cell -mediated immunity, while a Th2-mediated response is associated with the humoral immunity that if elevated IgE levels and eosinophilia. Prostaglandin (PG)E$_2$results in the decreased capability of Iymphocytes to produce Thl cytokines, with a shift toward a Th2 cytokine response. Chlorpyrifos (CPF) has been reported to impair the blastogenesis and response of T Iymphocytes. CPF also induces delayed febrile effects, which results from the activation of COX -PGE$_2$pathway. The purpose of this study is to determine the effort of CPF on the in vitro production of Th cytokines and the role of PGE$_2$on the CPF-induced production of Th cytokines. Splenocytes obtained from male BALB/c mice were pretreated with CPF(0.1, 1, 10 and 100$\mu$M) in the presence of absence of indomethacin or PGE$_2$for 12 h and then were incubated with concanavalin (Con) A for 48 h. These results showed that CPF remarkedly reduced the production of splenic interleukin (IL)-2 and interferon (IFN)-γ in a dose-dependent manner. CPF significantly increased the splenic IL-4 production at low doses (0.1 and 1$\mu$M) but did not affect at high doses (10 and 100 $\mu$M). Indomethacin reduced the CPF-decreased production of IL-2 and IFN-γ in a dose -dependent manner and significantly attenuated the production of IL-4 increased by CPF 0.1 $\mu$M. High dose of CPF significantly reduced the PGE$_2$-decreased production of IL-2 and IFN-γ, while the PGE$_2$- induced production of IL-4 was significantly enhanced by CPF 1 $\mu$M. These findings suggest that CPF nay down-regulate the immune response of Th 1 type by the suppressed production of IL-2 and IFN-γ, with a shift toward a Th2 cytokine response. The CPF-decreased production of Thl cytokines may not be mediated by endogenous PGE$_2$. Also, CPF may attenuate the exogenous PGE$_2$-decreased Th 1 immune response in a dose--dependent manner but may affect dose-independently the PGE$_2$-induced Th2 immune response.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.6
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• pp.892-897
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• 2005

Macrophage colony-stimulating factor (M-CSF) is a growth factor required for growth and differentiation of mononuclear phagocyte lineage. Total and 16 poly (A) mRNA of bovine M-CSF were isolated from healthy bovine peripheral mononuclear cells stimulated by phobol 12-myristste 13-acetate (TPA). The more compatible cultured mononuclear cells were 5${\times}$10/ml for RNA isolation. TPA-activated mononuclear cells increased the level of M-CSF-mRNA more than concanavalin A (Con A) and lipopolysaccharide (LPS). The optimal analysis of reverse transcriptase-polymerase chain reaction (RT-PCR) for14 Macrophage colonystimulating factor (M-CSF) as a growth factor required for bovine M-CSF was denaturation at 94$^{\circ}C$ for 1 minute, annealing at 57$^{\circ}C$ for 1 minute, extension at 72$^{\circ}C$ for 1 minute for 30 cycles. The size of cDNA of bovine M-CSF by RT-PCR was 774 base pairs. A 774 base pairs cDNA encoding bovine M-CSF was synthesized by reverse transcriptase polymerase chain reaction (RT-PCR). Ligated cDNA was transformed to competent cells and then plasmid isolation and digestion was performed. Molecular cloning and sequencing were performed for cDNA of bovine M-CSF. The size of cloned cDNA of bovine M-CSF was 774base pairs. The homology of base sequence and amino acid sequence was 88% and 86% compared with known human M-CSF, respectively. From a high degree of sequence similarity, the obtained cDNA of bovine M-CSF is thought be a specific gene of bovine M-CSF.

• Journal of Life Science
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• v.17 no.5 s.85
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• pp.694-698
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• 2007

Berberine, an alkaloid initially isolated from chinese herbal medicine, has antibiotic activities against a variety of organisms including bacteria, viruses, fungi, protozoans, and chlamydia. Furthermore, berberine has shown a number of beneficial effects, including anti-tumor, anti-inflammation, and vasodilatory effects. In this work we have investigated the effects of berberine on lymphocyte proliferation and GM-CSF production in mice. Mouse splenocytes were incubated with berberine and concanavalin A(Con A) to observe the effects on cell proliferation. The culture supernatants of splenocytes exposed to berberine, berberine plus LPS, and berberine plus Con A were harvested to assay GM-CSF. The cell proliferation of nice splenocytes exposed to berberine only($1{\mu}g/ml$) was increased significantly more than PBS(control) group. But the Con A-induced cell growth was inhibited by berberine. The GM-CSF production from mice splenocyte culture exposed to berberine only was increased in comparison with PBS(control) group, but the production of it with LPS or Con A was inhibited by berbeline. The present findings may explain lympocyte proliferating and regulating effects of berberine.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.131-136
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• 1994

In the present study, in order to make an in vitro model of gastric mucosa for physiological and pharmacological studies, we established two immortalized gastric surface mucous cell lines (GSM06 and GSM10), which produce periodic acid-Schiff (PAS)-and concanavalin A (Con A)-positive glycoproteins, from a primary culture of gastric fundic mucosal cells of adult transgenic mice harboring a temperature-sensitive simian virus 40 large T-antigen gene 〔1]. Gastric fundic mucosal cells were isolated as a modification of a previously described method for rats by Schepp et al. (2). The isolated gastric fundic mucosal cells were cultured in DME/F12 medium supplemented with 2% fetal bovine serum (FBS), 1% ITES (consisting of 2 mg/1 insulin, 2 mgg/1 transferrin, 0.122 mg/1 ethanolamine and 0.00914 mg/1 sodium selenite) and 10 ng/ml recombinant epidermal growth factor (EGF) in a collagen-coated culture dish. To remove fibroblastic cells from the culture, gastric mucosal cells were incubated in the culture medium containing dispase (25 U/ml) for 24 h. The cells, uncontaminated with fibroblastic cells, were then cloned by colony formation. In our series of three attempts, two cell lines (GSM06 and GSM10) have been established at last. The cells proliferated, attached to the dish ana grew until confluent monolayers were formed, and maintained tight contact with neighboring cells. Both GSM06 and GSM10 cells have now been in culture for more than 9 months with regular passaging. The either cell produced

• The Korean Journal of Physiology and Pharmacology
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• v.1 no.1
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• pp.91-96
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• 1997

Lipocortin 1 has been proposed as a putative mediator of anti-inflammatory actions of glucocorticoids. We investigated the role of lipocortin 1 in the effect of dexamethasone using rat splenic leukocytes. Concanavalin A(ConA; 1 ${\mu}g/ml$) increased the leukocyte proliferation and nitric oxide(NO) generation, which were measured as $[^3H]-thymidine$ uptake by the cells and nitrite accumulation in the culture media, respectively. Dexamethasone suppressed ConA-induced cell proliferation, in a concentration-dependent manner with $EC_{50}$ around 50nM. The addition of anti-lipocortin l(Anti-LCl) reversed dexamethasone effects: 0.24, 1.2, 6 ${\mu}g/ml$ of Anti-LC1 reversed dexamethasone(50 nM)-induced suppression of thymidine uptake by $9{\pm}3%$, $16{\pm}3%$, $36{\pm}5%$, respectively; 0.24, 1.2, and 6 ${\mu}g/ml$ of Anti-LCI reversed dexa-methasone-induced decrease of nitrite concentration by $49{\pm}16%$, $61{\pm}20%$, $77{\pm}19%$, respectively. The present data indicate that lipocortin 1 mediates, at least in part, glucocorticoids-induced suppression of leukocyte proliferation and blockade of NO generation.

• Korean Journal of Medicinal Crop Science
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• v.15 no.2
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• pp.132-137
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• 2007

Inhibitory effect of Scutellaria baicalensis ethanol extracts (SR) on chemical mediator release and immunoglobulin (Ig) production from Sprague-Dawley rats originated cells as type I allergic reaction was examined. SR showed concentration-dependent inhibition on basal and concanavalin A (ConA)-stimulated Ig production. In the mesenteric lymph node lymphocytes, the inhibitory effect of SR on the IgE production in the presence of Con A was stronger than these on IgA and IgG production. Moreover, tumor necrosis factor-alpha $(TNF-{\alpha})$ production-inhibiting effect of SR in the presence ConA was observed. However, SR did not affect the production of $interferon-{\gamma}$. SR also inhibited histamine release from the peritoneal exudate cells stimulated with a calcium ionophore A23187. In the case of leukotriene B4, SR markedly inhibited it at the concentration of 100 mg/ml. From these results, ethanol extracts obtained from Scutellaria baicalensis may have an anti-allergic effect on the intestinal system of rats.

• The Journal of Korean Medicine
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• v.32 no.3
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• pp.25-34
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• 2011

Objectives: We performed simultaneous determination of five constituents by HPLC in Samul-tang (SMT). Additionally, we investigated the immune-stimulatory potential of SMT on specific cellular and humoral immune responses in ovalbumin (OVA)-immunized mice. Methods: Reverse-phase chromatography using a Gemini C18 column operating at $40^{\circ}C$, and photodiode array (PDA) detection at 190-400 nm, were used for quantification of the five components of SMT. Mobile phase using a gradient flow consisted of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. C57BL/6 mice were immunized intraperitoneally with OVA/alum ($100{\mu}g/200{\mu}g$) on days 1, 8, and 15. The extract of SMT (1000 mg/kg) was given to mice orally for 21 days (from day 1 to day 21). At day 22, OVA-, lipopolysaccharide (LPS)- and concanavalin A (Con A)-stimulated splenocyte proliferation and OVA-specific and total antibodies were measured in plasma. Results: Calibration curves were acquired with $r^2$>0.9999, and the relative standard deviation (RSD, %) for intra- and inter-day precision were both less than 3.5%. The recovery was in the range of 95.69-115.12%, with an RSD less than 6.0%. The contents of five components in SMT were 1.08-15.30 mg/g. SMT significantly enhanced Con A-induced splenocyte proliferation in OVA-immunized mice (p<0.01). Also, SMT significantly enhanced OVAspecific IgG, IgG1 and total IgM levels in plasma compared with the OVA-immunized group. Conclusions: The established method will be applied for the quantification of major components and immunestimulating activity in OVA-immunized mouse model of SMT.

• The Journal of the Korean Society for Microbiology
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• v.10 no.1
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• pp.1-8
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• 1975

Despite a number of recent studies on appendix its function appears to remain unknown. The present studies were undertaken in order to extend and confirm the previous studies concerning the role of appendix in immune response. An early hemagglutinin response of mercaptoethanol sensitive antibody(IgM antibody) in rabbit injected intravenously(i.v.) with 200mcg of bovine gamma globulin(BGG) was abolished by lethal whole body irradiation(900 r), but preserved in animals whose appendix and bone marrow were shielded during irradiation. Late formation of mercaptoethanol resistant antibody(IgG antibody) and the development of memory in bone marrow shielded animals were not affected by irradiation of the appendix. Formation of either IgM or IgG antibody to sheep red blood cells(SRBC) injected i.v. as determined by direct plaque forming cell(DPFC) technique in spleen were effectively abolished by appendectomy, thymectomy, or both followed by irradiation. When bone marrow was shielded in combination with autologous appendix reconstitution, DPFC response was about 5 times greater than the sum of two. Lysed appendix cells failed to restore the response. Lethally irradiated rabbits restored with combination of autologous appendix and thymus cells showed DPFC responses which were essentially normal. Three pools of appendix were obtained by manual separation technique and were stimulated with soluble concanavalin A(Con A), phytohemagglutinin-P(PHA) and pokeweed mitogen(PWM). Rabbit appendix cells responded to Con A, PHA and PWM. Cells of thymus dependent area(TDA) of the appendix were relatively enriched in their response to T cell mitogens compared to dome and follicle cells. The PHA/Con A responsive ratio of appenix TDA subpopulation was high, indicating that Con A responsive cells have a wider distribution among appendix. This finding showed that interfollicular area of the appendix is thymus-dependent. The present studies confirmed other evidence that the rabbit appendix cells itself are unable to form antibody and T lymphocytes in appendix TDA may be heterogenous, and that the appendix cells are synergistic with either bone marrow or thymus cells in the early hemagglutinin on splenic antibody response to BGG or SRBC.