• The Plant Pathology Journal
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• v.19 no.5
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• pp.260-265
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• 2003

Gummy stem blight caused by Didymella bryoniae occurs exclusively in cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. In this study, cultural conditions for the mass-production of pycnidiospore by Metal Halide (MH) lamp irradiation were maximized. The mycelia were cultured at $26^{\circ}C$ on PDA for 2 days under dark condition, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$. Results show that a great number of pycnidia were simultaneously formed. The pycnidiospores produced were then used as immunogen. Fusions of myeloma cell (v-653) with splenocytes from immunized mice were carried out. Two hybridoma cell lines that recognized the immunogen D. bryoniae were obtained. One monoclonal antibody (MAb), Dbl, recognized the supernatant while another MAb, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid of the two MAb, Dbl and Db15, the immunotypes of which were identified as IgG1 and IgG2b, respectively. Titers of MAb Dbl and MAb Db15 were measured and the absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with D. bryoniae but none reacted with other viral isolates, Cucumber mosaic virus and Cucumber green mottle mosaic virus. Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{-3}$/well by indirect ELISA. Characterization of the MAbs Dbl, Db15 antigen by heat and protease treatments, which suggests that the epitope recognized by these two MAbs was glycoprotein.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.2
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• pp.428-435
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• 2006

Cortex Mori radicis has been used as a components of antidiabetics, antiasthma and diuresis in Oriental Medicine. This experiment examined the effect of an extracts, obtained from the acetone(CM-A) and aqueous(CM-W) extracts of Cortex Mori radicis, on hair growing activity of the C57BL/6N mice after topical application to skin. We investigated the number of hair follicle and mast cells, and changes of subpopulation of splenocytes and thymocytes in skin for 16 day. The results were as follows : The Hair growing effect in experimental groups was more increased in 85%(CM-A) and 90%(CM-W) than control group(10%) in hair depilated area. The number of hair follicle in experimental groups(CM-A and CM-W) was more increased than control group. Splenic B/T lymphocytes of CM-A group were decreased compare to control group. CD4/CD8 positive TH cells in splenic T lymphocytes of CM-W group were increased compare to control group. These results suggest that CM-A and CM-W may be used in treatment of alopecia areata.

• Korean Journal of Veterinary Research
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• v.34 no.4
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• pp.769-779
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• 1994

The root of Bupleurum falcatum L.(BF) has been widely used in oriental medicine as a major camponent in many prescriptions for chronic hepatitis, renal disease, tuberculosis and some other infectious diseases. Many attempts have done to investigate the therapeutic effects of these principles. However, any kinds of screenig on immune regulatory- and antitumor- effects of BF has not been reported. The present study, therefore, was undertaken to investigate the BF-effects on cellular- and humoral-immune responses, phagocytic activities of macrophages, lymphokine- and Immunoglobulin(Ig)-production of lymphocytes, tumorigenesis of implanted sarcoma 180 cells and B16 melanoma cells, and proliferations of some tumor cell lines(Fsa II, 3LL and EL4). BF increased phagocytic activities of mouse peritoneal macrophages in a dose- and time-dependent fashion. Arthus reaction and antibody responses to SRBC were slightly enhanced but delayed hypersensitivity was depresed when BF was injected before- and after-SRBC sensitization. BF inhibited the proliferative responses of human tonsillar lymphocytes to PHA- and Con A-stimulation but slightly augmented the response of these cells to Staphylococcus aureus Cowan 1(SAC)-activation. Ig secretion of human mononuclear cells activated with SAC was slightly increased by BF. BF significantly augmented the SAC-induced IL 6 production of human mononuclear cells but not influenced Con Ainduced IL 2 secretion. NK cell activities of mouse splenocytes were somewhat increased when BF was pretreated and this responses were due to the increment of binding affinities of effector cells to target cells and of lytic activities of effector cells against target cells. In vitro BF significantly inhibited the proliferations of cancer cells such as Fsa II, 3LL and EL4 strains. BF decreased not only the frequency of tumor induction but also the tumor size per sarcoma 180 or B16 cell-implanted mouse. Taken together, these results indicate that BF is one of the potential immunomodulator, and suggest its possibility to be used as a desirable antitumor agent.

• Korean Journal of Veterinary Research
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• v.55 no.1
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• pp.21-30
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• 2015

The immune system is specifically sensitive to oxidative stress induced by ionizing radiation because of its rapid proliferative activity. For this reason, an instructive immune system is one of the best ways to minimize side effects, such immunodeficiency, of gamma radiation. Over the past few decades, several natural plants with antioxidant and immunomodulatory properties have been identified as adjuncts for nontoxic and successful radiotherapy. Hizikia fusiforme extract (HFE) containing plentiful dietary fiber and fucoidan is known for its instructive antioxidant capacity, immunomodulation abilities, and immune activation. In this study, we determined whether HFE protects radiosensitive immune cells from gamma radiation-induced damage. C57BL/6 mice were irradiated with gamma-ray. The effect of HFE on the ionizing radiation damage of immune cells was then evaluated with an MTT assay, 3H-thymidine incorporation assay, and PI staining. We found that HFE stimulated the proliferation of gamma-ray irradiated immune cells without cytotoxic effects. We also observed that HFE not only decreased DNA damage but also reduced gamma radiation-induced apoptosis of the immune cells. Our results suggest that HFE can protect immune cells from gamma-ray damage and may serve as an effective, non-toxic radioprotective agent.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.8
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• pp.1182-1187
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• 2003

Two experiments involving 92 crossbred, 21 day old weaned pigs were used to evaluate the effect of glutamine supplement in a dietary or culture medium on lymphocyte proliferation. In Exp. 1, 88 pigs were fed diets supplemented with 0, 0.5, 1.0, or 1.5% glutamine for 28 days. Lymphocytes were prepared from peripheral blood mononuclear cells (PBMC), ileal Peyer's patches (PP), the mesenteric lymph node (MLN), and the spleen in each dietary supplement group on days 7, 14, or 28 postweaning. Lymphocytes were cultured at $37^{\circ}C$ for 72 h in a RPMI-1640 medium with or without mitogen-stimulated, and pulsed with 3Hthymidine for an additional 18 h. The stimulation index of PBMC proliferation in 1.0% dietary glutamine supplement group and both of the MLN and splenocytes proliferation in 1.5% dietary glutamine supplement group was significantly (p<0.05) increased at 14 days postweaning. In Exp. 2, four weaned pigs were fed a basal diet for 14 days. The 3H-thymidine incorporation of PBMC, PP, and MLN cells, incubated with 0.125 to 0.25 mM glutamine in culture medium were markedly enhanced with Con A-stimulated, however, the splenocyte proliferation was not affected in the addition of glutamine medium. These observations suggest that dietary glutamine supplement might enhance the lymphocyte proliferation of weaned pigs.

• BMB Reports
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• v.33 no.1
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• pp.92-95
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• 2000

The HIV-1 p24(202-221) sequence ETINNEEEWDRVHPV HAGP contains a B-cell epitope with the earliest immune response and the highest antibody titer against anti-mouse sera obtained by immunization with p24 antigens. A novel mouse monoclonal antibody (mAb) was generated against the immunodominant B-cell epitope of the HIV-1 p24 capsid protein, p24(202-221). BALB/c mice were immunized with the four branched multiple antigenic peptide (MAP) containing the HIV-1p24(202-221) sequence, and antibody-secreting hybridoma were produced by fusion of mouse splenocytes with P3X63Ag8.653, mouse myeloma cells. One clone which produced the antigen-specific mAb named KI-24 (Isotype IgG1, light chain: ${\kappa}$) was identified. mAb KI-24 was highly specific for both the p24(202-221) and p24 proteins when analyzed by ELISA and Western blotting. Since p24(202-221) also contains a cytotoxic T-lymphocyte epitope, this specfic peptide epitope and the monoclonal antibody with specific reactivity against the p24 protein and p24(202-221) can be used in peptide vaccine development and p24 antigen detection from HIV patients.

• BMB Reports
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• v.31 no.1
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• pp.58-63
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• 1998

In this study, a mouse monoclonal antibody (mAb) against gp41(584-618), the immunodominant epitope protein, was generated. For this purpose, BALB/c mice were immunized with double branched multiple antigenic peptides derived from the HIV-1 gp41(584-618) sequence, and antibody-secreting hybridoma were produced by fusion of mice splenocytes with SP2/0 myeloma cells. One clone producing an antigen specific mAb, termed KI-41(isotype IgG1) was identified, whose specific reactivity against gp41(584-618) could be confirmed by ELISA and Western blot analysis. Epitope mapping revealed the recognition site of the mAb KI-41 to be located around the sequence RILAVERYLKDQQLLG, which comprises the N-terminal region within the immunized gp41(584-618) peptied. Since this mAb recognizes this specific epitope within the HIV-1 gp41 without any cross-reactivity to other immunodominant regions in the HIV-2 gp35, KI-41 will provide some alternative possibilities in further applications such as the development of indirect or competitive ELISA for specific antibody detection in HIV-1 infection or for other basic researches regarding the role and function of HIV-1 gp41.

• Preventive Nutrition and Food Science
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• v.5 no.4
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• pp.219-224
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• 2000

Dieary effects of sea tangle on immune functions were investigated in diabetic mice. Four groups of ICR mice weighing 33.36$\pm$1.01 g were fed either an AIN-76 diet only (control), or with additional sea tangle powder, sea tangle water extract, and alginate at the level of 13.6%, 4%, and 1%, respectively by weight. Cellulose was omitted in sea tangle powder and alginate diets. After 10 days of feeding respective experimental diets, all mice were made diabetic by five consecutive intramuscular injections of streptozotocin (40mg/kg body weight per day) and fed the diets for four more weeks. Plasma IgG concentrations but not those of IgM were significantly higher in mice fed sea tangle powder, extract or alginate than those on the control diet. Plasma TNF$\alpha$ levels were, however, lower in those fed sea tangle power or water extract than control and alginate fed groups. TNF$\alpha$ releases from macro phages isolated from four groups and cultured with 5 $\mu\textrm{g}$/mL LPS for 24 hors showed a similar tendency to the results of plasma concentrations in the respective groups, but IL-1$\beta$ releases were not different among four groups. Lymphocyte proliferation in response to LPS (10$\mu\textrm{g}$/mL) measured using splenocytes cultured for 3 days was highest in the alginate fed group (594$\pm$38%) and those of sea tangle powder (536$\pm$47%) and extract (547$\pm$34%) fed groups tended to be higher than the control (523$\pm$30%). It is concluded that sea tangle contains immunomodulatory components besides alginate that could enhance humoral immunity of itself. The immunomodulatory effects of sea tangle constituents is regarded as beneficial for diabetic subjects.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.2
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• pp.987-991
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• 2013

Aim: To study anti-tumor effects of exosomes from class II transactivator (CIITA) gene transfected CT26 cells. Methods: In this study, we established an MHC class II molecule-expressing murine colon cancer cell line (CT26-CIITA) by transduction of the CIITA gene. Immune effects in vitro and tumor protective results in vivo were tested and monitored. Results: Exosomes from CT26-CIITA cells were found to contain a high level of MHC class II protein. When loaded on dendritic cells (DCs), exosomes from CT26-CIITA cells significantly increased expression of MHC class II molecules, CD86 and CD80, as compared to exosomes from CT26 cells. In vitro assays using co-culture of immunized splenocytes and exosome-loaded DCs demonstrated that CIITA-Exo enhanced splenocyte proliferation and IFN-${\gamma}$ production of CD4+T cells, while inhibiting IL-10 secretion. In addition, compared to exosomes from CT26 cells, CT26-CIITA-derived exosomes induced higher TNF-${\alpha}$ and IL-12 mRNA levels. A mouse tumour preventive model showed that CT26-CIITA derived exosomes significantly inhibited tumour growth in a dose-dependent manner and significantly prolonged the survival time of tumour-bearing mice. Conclusion: Our findings indicate that CT26-CIITA-released exosomes are more efficient to induce anti-tumour immune responses, suggesting a potential role of MHC class II-containing tumour exosomes as cancer vaccine candidates.

• Nutritional Sciences
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• v.9 no.2
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• pp.68-73
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• 2006

This study was aimed at investigating hepatic toxicity and immunomodulating effects of defatted methanol extracts of two kinds of medicinal plants, Rubus coreanus Miq. and Atractylodes japonica Koidz. in mice. Defatted methanol extracts of fruits of Rubus coreanus Miq. and rhizome of Atractylodes japonica Koidz. were added at the level of 0.5% or 5%(w/w) to cholesterol-supplemented AIN-76 diet. Each diet was fed to 8 ICR male mice for 30 days. Weight gain and food efficiency ratio of the mice fed 5.0% extract of Rubus coreanus Miq. were significantly lower than those of the mice fed 0.5% extract Relative liver weight and activity of plasma alanine aminotransfernse were significantly increased only in the mice fed 5% extract of Atractylodes japonica Koidz. compared with the others. Splenocyte proliferation was not significantly different between the groups fed 0.5% or 5.0% extract of Rubus coreanus Miq. However, splenocyte proliferation was significantly decreased in the mice fed 5.0% extract of Atractylodes japonica Koidz. compared with that in the mice fed 0.5% Production of interleukin-2 by splenocytes from the mice fed 0.5% extract of Atractylodes japonica Miq. was significantly higher than the control value and it became lower with 5.0% dietary level. Secretion of $interferon-\gamma$ was not significantly different among groups. In conclusion, the defatted methanol extract of Atractylodes japonica Koidz. was likely to exert immunomodulating effect at the level of 0.5% but it may exert adverse effects on immune and liver functions at the level of 5.0%.