• Archives of Pharmacal Research
• /
• 제20권5호
• /
• pp.396-403
• /
• 1997

In order to direct the form of the immune response in an antigen-specific manner, we constructed a fusion protein (OVA/IL12) that contained the T cell-dependent antigen, ovalbumin (OVA), covalently linked to murine interleukin-12 (IL-12). The OVA/IL12 protein was produced in a baculovirus expression system and was purified by anti-OVA immunoaffinity chromatography. The purified OVA/ILI2 protein displayed potent IL-12 bioactivity in an IL-12 proliferation assay. BALB/c mice immunized with the OVA/IL12 protein produced increased quantities of anti-OVA IgG2a antibody compared with mice immunized with recombinant OVA alone. Lymph node cells from the immunized mice with the OVA/IL12 protein produced large amounts of IFN-,Y when restimulated in vitro with OVA, while those from mice immunized with the OVA protein produced little or no IFN-.gamma.. In contrast, immunization with a mixture of OVA and free recombinant IL-12 also induced IFN-.gamma. production, which was not OVA-specific. These studies indicate that the OVA/IL12 fusion protein can induce OVA-specific, Th1-dominated immune responses, and that the covalent linkage of OVA and IL-12 confines the effect of IL-12 to OVA-specific cells.

• Korean Journal of Acupuncture
• /
• 제22권2호
• /
• pp.107-125
• /
• 2005

Objectives : The aim of this study was to investigate the asthma-suppressive and immuno-regulatory effect of NR-HA(Notopterygii Rhizoma Herbal-acupuncture) at Pyesu(BL13) on OVA(ovalbumin)-induced asthma in mice. Methods : C57BL/6 mice out of all the experimental sloops, except the Normal group and the NR-HA group, were sensitized and challenged with OVA. The mice in the NR-HA group and the OVA-NR-HA group were treated with NR-HA(1%) at Pyesu(BL13). The mice in the OVA-Saline group were injected with saline at Pyesu(BL13). The mice in the OVA-Needle-prick group were treated with a single prick with an injection needle at Pyesu(BL13). NR-HA, saline injection and needle prick were administered for 8 weeks, three times a week Results : in vitro 1. The populations of granulocytes, $CD3e^-/CCR3^+$cells, $CD69^+/CD3e^+$ cells, $CD4^+\;cells\;and\;CD23^+/B220^+$ cells in the OVA-induced asthmatic mouse lungs decreased significantly by NR-HAS(Notopterygii Rhizoma Herbal-acupuncture solution). 2. The lung weight and total cells in lung of the OVA-NR-HA group decreased significantly compared with those of the OVA-Control group. 3. Total leukocytes and eosinophils in BALF of the OVA-NR-HA group decreas ed significantly compared with those of the OVA-Control group. 4. The collagen accumulation in the lung sections of the OVA-NR-HA group decreased significantly compared with that of the OVA-control group. 5. The concentrations of IL-4, IL-5, IL-13, IgE in BALF and serum of the OVA-NR-HA group decreased significantly compared with those of the OVA- Control group. 6. The numbers of $Gr-1^+/CD11b^+,\;CCR3^+,\;CD3e^+, \;CD19^+,\;CD3e^+/CD69^+$ cells in the OVA-NR-HA group decreased significantly compared with those of the OVA-Control group. 7. The mRNA expressions of $TNF-{\alpha}$, IL-5, IL-4 and IL-13 in lung of the OVA-NR-HA group decreased significantly compared with those of the OVA- Control group. 8. The NR-HA group did not show my considerable difference from the Normal group. The OVA-saline group and the OVA-Needle prick group showed suppressive effects on OVA-induced asthma however they were not statistically significant. Conclusion : These results suggest that NR-HA at Pyesu(BL13) is considered to be effective in treating asthma and to be put to practical use in the future asthma clinic.

• 한국수정란이식학회지
• /
• 제10권3호
• /
• pp.203-208
• /
• 1995

This study was carried out to efficiently use the ultrarapid freezing method in the cryopreservation of mouse ova. For this, the effects of dehydration method, oval vigour and $0^{\circ}C$ controlling method on post-thawing viability were investigated. Fresh mouse ova were dehydrated in mPBS with 3.5M DMSO and /or 0.25M sucrose, and directly immersed in L$N_2$ for ultrarapidly freezing. The frozen ova were thawed at 37$^{\circ}C$, rehydrated in mPBS with 0.25M sucrose, and then repeatedly washed in HAM's Fl0 before evaluating the morphological normality of frozen-thawed ova. The results obtained showed that there was difference between treatments in a experiment. 1) The post-thawing viability of ova dehydrated in multi-step (48.4$\pm$13.8%) was higher than that of ova in two-step (40.9$\pm$14.0%). 2) The post-thawing viability of fertilized ova (87$\pm$14.0%) was significantly(p<0.0l) higher than that of unfertilized ova (5.4$\pm$5.4%). 3) The post-thawing viability of ova dehydrated and rehydrated using a cooling machine (95.8$\pm$4.2%) was significantly(p<0.05) higher than that on ice(84.1$\pm$9.9). In conclusion, in order to efficiently cryopreserve ova in vitro with ultrarapidly freezing method, highly viable embryos should be selected, heavy osmotic shock to the dehydrating ova should be avoided, and embryos in high osmotic condition were dehydrated and rehydrated in a constantly low temperature.

• 한국축산식품학회지
• /
• 제36권1호
• /
• pp.14-18
• /
• 2016

Oral administration of soluble antigen can induce peripheral tolerance to the antigen. This study was conducted to evaluate whether gamma-irradiated ovalbumin (OVA) can induce oral tolerance. To investigate this, we administrated intact or irradiated OVA to mice, induced allergic response using intact OVA and alum, then compared humoral and cellular immune responses. Mice treated with gammairradiated OVA had less OVA-specific IgE compared with those who were administered intact OVA. There was no difference in levels of OVA-specific IgG+A+M, IgG1, and IgG2a. Splenocytes of mice administered irradiated OVA showed similar OVA-specific T cell proliferation and secretion of IFN-γ and IL-4. However, there was an increase in IL-2 and a decrease of IL-6 secretion in mice treated with irradiated OVA. These results indicate that gamma-irradiated OVA have similar effects to intact OVA on antigen tolerance.

• Journal of Acupuncture Research
• /
• 제33권1호
• /
• pp.23-36
• /
• 2016

Objectives : The aim of this study was to investigate the asthma-suppressive and immuno-regulatory effect of AHCR-HA (Root of Asarum sieboldii Miq. water extract) on OVA (ovalbumin)-induced asthma in mice. Methods : All C57BL/6 mice experimental groups, except the Normal and N-AHCR groups, were sensitized with OVA. The mice in the N-AHCR group and the OVA-AHCR group were treated with water extract of AHCR (1 %) by an oral administration, and the OVA-Saline group were treated with saline solution. Oral dosesof AHCR water extract and saline were administered for 8 weeks, three times a week. Results & Conclusion : The lung weight and total cells in the lungs of the OVA-AHCR group decreased significantly compared with those of the OVA-Saline group. Total leukocytes and eosinophils in BALF and photomicrographs of the OVA-AHCR group decreased significantly compared with those of the OVA-Saline group. The collagen accumulation in the lung sections of the OVA-AHCR group decreased significantly compared with those of the OVA-Saline group. The concentrations of IL-4, IL-5, IL-13 and IgE in BALF and the serum of the OVA-AHCR group decreased significantly compared with those of the OVA-Saline group. The mRNA expressions of $TNF-{\alpha}$, $IL-1{\beta}$, IL-4, IL-5 and IL-13 in the lungs of the OVA-AHCR group decreased significantly compared with those of the OVA-Saline group.

• Journal of Acupuncture Research
• /
• 제24권3호
• /
• pp.39-62
• /
• 2007

Objectives : The aim of this study was to investigate the asthma-suppressive and immuno-regulatory effect of Lepidii Semen Herbal-acupu ncture(LS-HA) at $Joksanmi(ST_{36})$ on ovalbumin(OVA)-induced asthma in mice. Methods : C57BL/6 mice out of all the experimental groups, except the Normal group and the LS-HA group, were sensitized and challenged with OVA. The mice in the LS-HA group and the OVA-LS-HA group were treated with LS-HA(1%) at $Joksanmi(ST_{36})$. The mice in the OVA-Saline group were injected with saline at $Joksanmi(ST_{36})$. The mice in the OVA-Needle-Prick group were treated with a single prick with an injection needle at $Joksanmi(ST_{36})$. LS-HA, saline injection and needle prick were administered for 8 weeks, three times a week. Results : The lung weight and total cells in lung of the OVA-LS-HA group decreased significantly compared with those of the OVA-Control group. Total leukocytes and eosinophils in BALF of the OVA-LS-HA group decreased significantly compared with those of the OVA-Control group. The collagen accumulation in the lung sections of the OVA-LS-HA group decreased significantly compared with that of the OVA-control group. The concentrations of IL-4, IL-5, IL-13, IgE in BALF and serum of the OVA-LS-HA group decreased significantly compared with those of the OVA-Control group. The numbers of $Gr-1^+/CD11b^+$, $CCR3^+$, $CD3e^+$, $CD19^+$, $CD3e^+/CD69^+$cells in the OVA-LS-HA group decreased significantly compared with those of the OVA-Control group. The mRNA expressions of $TNF-{\alpha}$, IL-5, IL-4 and IL-13 in lung of the OVA-LS-HA group decreased significantly compared with those of the OVA-Control group. Conclusions: These results suggest that LS-HA at $Joksamni(ST_{36})$ is considered to be effective in treating asthma and to be put to practical use in the future asthma clinic.

• Archives of Pharmacal Research
• /
• 제21권5호
• /
• pp.537-542
• /
• 1998

to more effectively drive immune responses toward antigen-specific T helper type 2 (Th2) cell-mediated responses, we constructed a mammalian expression vetor (oPVA/IL4) carrying a fused gene in which the ovalbumin (OVA) cDNA was covalently linked to murine interleukin-4 (IL-4) cDNA. A biologically active OVA/IL4 DNA, as demonstrated by Wes tern blotting and cytokine bioassay. In tramuscular injection of BALB/c mice with the pOVA/IL4 DNA increased both the production of OVA-specific IL-4 by CD$4^{+}$ T cells and the ratio of anti-OVA lgG1 to anti-OVA lgG2a isotypes, while the injection with the pOVA DNA alone, or with the mixture of the pOVA and pIL4 DNA did no or little increase. furthermore, the OVA-specific, Th2 cell-mediated immune responses were significantly enhanced by multiple injections with the pOVA/IL4 DNA. These studies indicate that the direct linkage of an OVA gene to an IL-4 gene in the expression plasmid confines the effects of IL-4 to the OVA-specific cells, efficiently driving the immune response toward OVA-specific, Th2 cell-mediated responses.

• Asian-Australasian Journal of Animal Sciences
• /
• 제26권4호
• /
• pp.545-551
• /
• 2013

This study aimed to present a mouse model of ovalbumin (OVA) induced allergic diarrhea under a sub-barrier system and investigate the development of gut microbiota in this model. Male BALB/c mice were systemically sensitized with OVA or sham-sensitized with saline, and followed by oral OVA intubation, leading to OVA-specific acute diarrhea. Compared with sham-sensitized mice, sera OVA-specific IgG1 and total IgE in OVA-sensitized mice were dramatically elevated, and the number of mast cells was greatly increased in the jejunum of the OVA-sensitized mice. Principle component analysis of the DGGE profile showed that samples from group of OVA-sensitized mice and group of sham-sensitized mice were scattered into two different regions. Real-time PCR analysis showed that the number of 16S rRNA gene copies of Lactobacillus in the colon of OVA-sensitized mice decreased significantly, while there was no significant difference in the number of Bifidobacterium and total bacteria. In conclusion, OVA-specific allergic diarrhea was successfully induced under a sub-barrier system, and changes of allergic reactions during induction was coupled with changes in gut microbiota, especially the number of colonic Lactobacillus, but the role of gut microbiota in the development of food allergic reactions needs to be further evaluated.

• 동의생리병리학회지
• /
• 제29권6호
• /
• pp.467-474
• /
• 2015

Allergic rhinitis is a growing tendency to increase. Following this tendency, allergic rhinitis is lively studied also in Korean oriental medicine. Bupleuri Radix (BR) has been used for many inflammatory diseases, but experimental backgrounds are not enough to treat allergic rhinitis. So in this study, effects of BR on OVA-induced allergic rhinitis model of BALB/c mice were examined. Thirty two BALB/c mice are equally devided into four groups; control group, OVA group, OVA+BR group, OVA+Cet group. The OVA, OVA+BR and OVA+Cet groups were induced allergic rhinitis by sensitizing to OVA. And then saline solution included BR (10.6 mg/kg body weight) was administered to the OVA+BR group orally. The number of nasal rubbing and nasal sneezing was evaluated for 10 days and later serological and histological changes were analyzed. Serological analysis included the serum levels of cytokines and chemokines (IL-4, IL-5, IL-12, MCP-1, MIP-2), total IgE and OVA-specific IgE levels in serum. Histological analysis included thickness of nasal septum, eosinophil counts changes of nasal mucosa, infiltration of eosinophil in nasal mucosa and histological changes of nasal mucosa. The number of nasal rubbing and nasal sneezing was significantly decreased in the OVA+BR group. The serum levels of IL-4, IL-5, MCP-1, MIP-2 were significantly decreased in the OVA+BR group but the serum levels of IL-1β had not significance. Total IgE and OVA-specific IgE levels in serum were decreased in the OVA+BR group, but total IgE levels in serum had only significance. Thickness of nasal septum, eosinophil counts of nasal mucosa and infiltration of eosinophil in nasal mucosa were significantly decreased in the OVA+BR group. From the results of this study, we think that BR has an effect on improvement of allergic rhinitis by improving nasal rubbing and nasal sneezing, reducing histological changes of nasal mucosa and infiltration of eosinophil in nasal mucosa, inhibiting increasing of the serum levels of cytokines, chemokines and total IgE.

• 대한본초학회지
• /
• 제26권4호
• /
• pp.83-88
• /
• 2011

Objectives : The aim of this study was to investigate the asthma-suppressive and immuno-regulatory effect of Notopterygii Rhizoma(NR) extract on OVA(ovalbumin)-induced asthma in mice. Methods : C57BL/6 mice out of all the experimental groups, except the Normal group and the NRI group, were sensitized and challenged with OVA. C57BL/6 mice were exposed to OVA three times a week for 12 weeks and analyzed by flow cytometer, ELISA, H&E stain. Results : The concentrations of IL-4, IL-5, IL-13, IgE in serum of the OVA-NRII group decreased significantly compared with those of the OVA-Control group. The number of $Gr-1^+/CD11b^+$, $CCR3^+$, $CD3^+/CD19^+$, $CD3e^+/CD69^+$cells in the OVA-NRI group decreased significantly compared with those of the OVA-Control group. The collagen accumulation in the lung sections of the OVA-NRII group decredased signi- ficantly compared with that of the OVA-Control group. Conclusions : These results suggest that Notopterygii Rhizoma(NR) would be a effective candidate for herbal-originated anti-asthmatic drug. However, this drug should be further studied for characterization of the accurate action and underlying mechanism using variant disease model in the future.