• IMMUNE NETWORK
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• v.13 no.1
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• pp.30-33
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• 2013

Previously we showed that biodegradable nanoparticles containing poly-IC or CpG oligodeoxynucleotide (ODN) together with ovalbumin (OVA) were efficient at inducing MHC-restricted presentation of OVA peptides in dendritic cells. The CTL-inducing activities of the nanoparticles were examined in the present study. Nanoparticles containing poly-IC or CpG ODN together with OVA were prepared using biodegradable polymer poly(D,L-lactic acid-co-glycolic acid), and then were opsonized with mouse IgG. The nanoparticles were injected into the tail vein of mice, and 7 days later the OVA-specific CTL activities were measured using an in vivo CTL assay. Immunization of mice with the nanoparticles containing poly-IC or CpG ODN together with OVA elicited potent OVA-specific CTL activity compared to those containing OVA only. In accordance with these results, nanoparticles containing poly-IC or CpG ODN together with OVA exerted potent antitumor activity in mice that were subcutaneously implanted with EG7.OVA tumor cells. These results show that encapsulation of poly-IC or CpG ODN together with antigen in biodegradable nanoparticles is an effective approach for the induction of potent antigen-specific CTL responses in vivo.

• IMMUNE NETWORK
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• v.16 no.4
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• pp.233-241
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• 2016

DCs, like the sensory neurons, express vanilloid receptor 1 (VR1). Here we demonstrate that the VR1 agonists, capsaicin (CP) and resiniferatoxin (RTX), enhance antiviral CTL responses by increasing MHC class I-restricted viral antigen presentation in dendritic cells (DCs). Bone marrow-derived DCs (BM-DCs) were infected with a recombinant vaccinia virus (VV) expressing OVA (VV-OVA), and then treated with CP or RTX. Both CP and RTX increased MHC class I-restricted presentation of virus-encoded endogenous OVA in BM-DCs. Oral administration of CP or RTX significantly increased MHC class I-restricted OVA presentation by splenic and lymph node DCs in VV-OVA-infected mice, as assessed by directly measuring OVA peptide SIINFEKL-K^b complexes on the cell surface and by performing functional assays using OVA-specific CD8 T cells. Accordingly, oral administration of CP or RTX elicited potent OVA-specific CTL activity in VV-OVA-infected mice. The results from this study demonstrate that VR1 agonists enhance anti-viral CTL responses, as well as a neuro-immune connection in anti-viral immune responses.

• The Journal of Korean Medicine
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• v.32 no.3
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• pp.10-17
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• 2011

Objective: To evaluate the immune-stimulatory potential of extracts of Broussonetia kazinoki Siebold (BK) on specific cellular and humoral immune responses in ovalbumin (OVA)-immunized mice. Material and Methods: C57BL/6 mice were immunized intraperitoneally with OVA/alum ($100{\mu}g/200{\mu}g$) on days 1, 8, and 15. BK (100, 300 or 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. Further, the effects of BK on expression of cytokine mRNA in OVA-immunized mice splenocytes were evaluated by RT-PCR analysis. Results: BK significantly enhanced OVA-, LPS-, and Con A-induced splenocyte proliferation in OVA-immunized mice (p<0.01). BK also significantly enhanced total IgM and OVA-specific IgG1 levels in plasma compared with the OVA control group. Moreover, BK up-regulated significantly the expression of mRNA level of IL-2 and IFN-${\gamma}$ in splenocytes. Conclusions: BK has immune-stimulating activity in an OVA-immunized mouse model system, enhancing the Th1 immune response. BK showed no cytotoxicity in this system, suggesting that BK may be a safe and effective adjuvant in humans.

• Clinical and Experimental Pediatrics
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• v.53 no.4
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• pp.481-487
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• 2010

Purpose: Recently many studies show early exposure during childhood growth to endotoxin (lipopolysaccharides, LPS) and/or early exposure to allergens exhibit important role in development of allergy including bronchial asthma. The aim of this study was to evaluate the role of endotoxin and allergen exposure in early life via the airways in the pathogenesis of allergic airways inflammation and airway hyperresposiveness (AHR) in mouse model of asthma. Methods: Less than one week-old Balb/c mice was used. Groups of mice were received either a single intranasal instillation of sterile physiologic saline, 1% ovalbumin (OVA), LPS or $1.0{\mu}g$ LPS in 1% OVA. On 35th day, these animals were sensitized with 1% OVA for 10 consecutive days via the airways. Animals were challenged with ovalbumin for 3 days on 55th days, and airway inflammation, hyperresponsiveness, and cytokine expression were assessed. Measurements of airway function were obtained in unrestrained animals, using whole-body plethysmography. Airway responsiveness was expressed in terms of % enhanced pause (Penh) increase from baseline to aerosolized methacholine. Lung eosinophilia, serum OVA-IgE and bronchoalveolar lavage (BAL) fluid cytokine levels were also assessed. ANOVA was used to determine the levels of difference between all groups. Comparisons for all pairs were performed by Tukey-Kramer honest significant difference test; $P$ values for significance were set to 0.05. Results: Sensitized and challenged mice with OVA showed significant airway eosinophilia and heightened responsiveness to methacholine. Early life exposure of OVA and/or LPS via the airway prevented both development of AHR as well as bronchoalveolar lavage fluid eosinophilia. Exposure with OVA or LPS also resulted in suppression of interleukin (IL)-4, 5 production in BAL fluid and OVA specific IgE in blood. Conclusion: These results indicate that antigen and/or LPS exposure in the early life results in inhibition of allergic responses to OVA in this mouse model of astham. Our data show that early life exposure with OVA and/or LPS may have a protective role in the development of allergic airway inflammation and development of allergen-induced airway responses in mouse model of asthma.

• Korean Journal of Animal Reproduction
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• v.16 no.3
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• pp.199-208
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• 1992

These studies were carried out to investigate optimal physological conditions for in vitro fertilization (IVF) of mouse ova. The unfertilized ova were obtained by superovulation from ICR mice of 4 to 6 weeks old. Tyrode's 280 solution was used as basal media, and pH and osmolality of basal media were adjusted with the supplementation of sodium bicarbonate and sodium chloride, respectively. The optimal pH, and osmolality of culture media and the optimum period of sperm preincubation were examined in fertilization in vitro of mouse ova and the subsequent culture in vitro of embryos. The pH range of media examined was designed from 6.5 to 7.5 with 0.2 interval and the range of osmolality from 250 to 370 mOsm with 20 interval, and the period of sperm preincubation examined was 30, 60, 120, and 180 minutes. The ova developed to 2-cell embryosafter 26hrs. of incubation with preincubated sperm were evaluated as in vitro fertilized ones. The results obtained were summarized as follows: 1. The percentage of in vitro fertilized ova was highest (64.7%) in media of pH 7.1 and lowest (38.0%) in pH 6.7. No significant difference in % fertilized ova was found from the media of pH 7.1 to 7.5. Compared with the result from pH 7.1 medium, the pollyspermy was increased signifciantly (p<0.05) in the media of pH over 7.5 and below 6.9;, and the % degenerated ova was significantly (p<0.05) increased in the media of pH below 6.9. 2. The percentage of in vitro fertilized ova was highest (69.4%) in media of osmolality 330 mOsm and lowest (47.9%) in osmolality 250 mOsm. No significant difference in % fertilized ova was found from the media of osmolality 310 to 350 mOsm. Compared with the result from osmolality 330 mOsm in medium, the polyspermy aws increased significantly(p<0.05) in the media of osmolality over 350 mosmol and blow 290 mOsm, and the % degenerated ova was significantly (P<0.05) increased in the media of osmolality below 290 mOsm. 3. The percentate of in vitro fertlilized ova was highest (62.7%) in media of period sperm preincubation 180 min. and lowest (40.4%) in sperm preincubation 30 minutes. No significant difference in % fertilized ova was found from the media of sperm preincubation 120 to 180 minutes. Compared with the result from sperm preincubation 180 minutes in medium, the polyspermy was low differ no significantly(P<0.05) in the media of period sperm preincubation, and the % degenerated ova was signifciantly(P<0.05) increased in the media of sperm presincubation below 60 minutes.

• Korean Journal of Acupuncture
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• v.22 no.3
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• pp.119-135
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• 2005

Objectives : The aim of this study was to investigate the Asthma-suppressive and Immune-regulatory effect of AF-HA(Aristolochiae Fructus Herbal-acupuncture) at Joksamni(St36) in OVA(ovalbumin) induced asthma mouse model. Methods : C57BL/6 mice were sensitized and challenged with OVA(ovalbumin) for 12 weeks(once a week). The mice in the OVA-AF-HA group were treated with AF-HA at St36 for the later 8weeks(3times/week). The mice in the OVA-Needle-prick group were treated with single prick with an injection needle at St36 for the later 8 weeks(3times/week). Results : 1. The lung weight and the total cells in lung of the mice treated with AF-HA at St36 decreased significantly compared with those of the OVA-control group. 2. Total leukocytes and eosinophils in BALF of the mice group treated with AF-HA at St36 decreased remarkably compared with those of the OVA-control group. 3. The collagen accumulation in lung of OVA-AF-HA group decreased significantly compared with that of the OVA-control group, 4. The concentrations of IL-4, IL-5, IgE in BALF, and IL-4, IL-5, IL-13 in serum of the mice group treated with AF-HA at St36 decreased significantly compared with those of the OVA-control group. 5. The numbers of $Gr-1^+/CD11b^+\;and\;CD11b^+$ cells in lung of the mice group treated with AF-HA at St36 decreased significantly compared with those of the OVA-control group. 6. The numbers of $CCR3^+,\;CD4^+,\;CD8^+\;and \;CD3e^+/CD69^+$ cells in lung of the mice group treated with AF-HA at 5136 decreased significantly compared with those of the OVA-control group. 7. The mRNA expressions of $TNF-{\alpha}$, IL-4, IL-5, IL13 in lung of the mice group treated with AF-HA at St36 decreased significantly compared with those of the OVA-control group. Conclusion : These results suggest that Aristolochiae Fructus Herbal-acupuncture at Joksamni(St36) may be an effictive therapeutic method to treat asthma.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.5
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• pp.848-853
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• 2001

This research was conducted to study the effects on antigenicities (allergenicity) and structural changes of gamma-irradiated hen’s egg albumin (ovalbumin, OVA) by heating. Three groups of OVA solution (2.0 mg/mL) were prepared; 1) heat treatment; 2) irradiation after heating; 3) heating after irradiation. Samples were isothermally heated and/or irradiated at the absorption dose of 10 kGy. Competitive indirect ELISA was individually formatted with egg-allergic patients IgE (P-IgE), and mouse murine monoclonal IgG (M-IgG) and rabbit polyclonal IgG (R-IgG) for evaluating bindinhg abilities of antibodies to OVA in the sample solutions. Binding abilities of antibodies to thermally denatured OVA were changed : R-IgG to the sample treated with above 6$0^{\circ}C$, M-IgG to that above 7$0^{\circ}C$, and P-IgE to that above 8$0^{\circ}C$, respectively. P-IgE did not well recognize OVA heated at 8$0^{\circ}C$ and the above. However, binding abilities of M-IgG and R-IgG highly in creased. Significant differences of binding abilities were not observed in all samples with the combination of heat treatment and irradiation, regardless the order of the treatment. Turbidity of samples in creased both by heating and by irradiation, and the increase by irradiation was much higher than by heating. These results showed that allergenicity of OVA reduced by gamma irradiation was not affected by heating.

• Korean Journal of Food Science and Technology
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• v.43 no.1
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• pp.72-76
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• 2011

The adjuvant effects of Korean mistletoe lectin-C (KML-C) were investigated following the oral administration of KML-C with ovalbumin (OVA) as an antigen. Mice were orally immunized with OVA alone or admixed with various doses of KML-C or cholera toxin (CT), and the titer of OVA-specific antibody in the serum and mucosal secretions were determined. OVA+KML-C-treated mice showed high titers of IgA specific to CT in mucosal secretions. The antibody titers in the serum of OVA+KML-C-treated mice were comparable to those in the serum of OVA+CT-treated mice. When mice were immunized with OVA+KML-C or with CT alone and subsequently injected with OVA on the footpads after the primary immunization, they showed a more significant increase in delayed-type hypersensitivity reactions than when they were administered CT alone. These results suggest that KML-C is a potent immunoadjuvant that enhances both humoral and cellular immunity by the mucosal immune system.

• Korean Journal of Food Science and Technology
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• v.44 no.6
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• pp.759-762
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• 2012

Egg allergies have been reported as one of the most prevalent food hypersensitivities in the pediatric population. One of the major egg allergens is ovalbumin (OVA), which is the major protein in the egg whites. Phenethyl isothiocyanate (PEIC) from cruciferous vegetables has an effect on anti-inflammatory therapy. In the present report, we show that PEIC inhibits the nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activation induced by OVA. PEIC also inhibits the OVA-induced inducible nitric oxide synthase (iNOS) expression and nitrite production. However, PEIC did not suppress the cyclooxygenase-2 (COX-2) expression induced by OVA. These results suggest that PEIC has the specific mechanism for anti-inflammatory responses and efficient anti-allergic activities.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.7
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• pp.1169-1174
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• 2004

This study was carried out to assess the effects of electron beam (EB) radiation on the conformational changes of ovalbumin (OVA), based on the early works using gamma irradiation. The applied doses of OVA used were 3,5,7, and 10 kGy, respectively. The conformational alterations were measured with SDS-PAGE, GPC-HPLC, and competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA) using monoclonal anti-OVA IgG antibody. Irradiation caused a degradation and/or an aggregation of OVA molecule. Immunochemical structures of irradiated OVA were altered by irradiation. Effects of gamma and electron beam radiation were similar at the same absorbed doses. These results may be used for inhibition of food allergy and development of immunogen with EB radiation.