• Molecules and Cells
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• v.42 no.12
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• pp.869-883
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• 2019

Interleukin (IL)-15 is an essential immune-modulator with high potential for use in cancer treatment. Natural IL-15 has a low biological potency because of its short half-life and difficulties in mass-production. IL-15Rα, a member of the IL-15 receptor complex, is famous for its high affinity to IL-15 and its ability to lengthen the half-life of IL-15. We have double-transfected IL-15 and its truncated receptor IL-15Rα into CT26 colon cancer cells to target them for intracellular assembly. The secreted IL-15:IL-15Rα complexes were confirmed in ELISA and Co-IP experiments. IL-15:IL-15Rα secreting clones showed a higher anti-tumor effect than IL-15 secreting clones. Furthermore, we also evaluated the vaccine and therapeutic efficacy of the whole cancer-cell vaccine using mitomycin C (MMC)-treated IL-15:IL-15Rα secreting CT26 clones. Three sets of experiments were evaluated; (1) therapeutics, (2) vaccination, and (3) long-term protection. Wild-type CT26-bearing mice treated with a single dose of MMC-inactivated secreted IL-15:IL-15Rα clones prolonged survival compared to the control group. Survival of MMC-inactivated IL-15:IL-15Rα clone-vaccinated mice (without any further adjuvant) exceeded up to 100%. This protection effect even lasted for at least three months after the immunization. Secreted IL-15:IL-15Rα clones challenging trigger anti-tumor response via CD4⁺ T, CD8⁺ T, and natural killer (NK) cell-dependent cytotoxicity. Our result suggested that cell-based vaccine secreting IL-15:IL-15Rα, may offer the new tools for immunotherapy to treat cancer.

• Journal of Pharmacopuncture
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• v.4 no.1
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• pp.5-21
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• 2001

The influence of electroacupuncture (EA), a traditional Chinese medical treatment, on type Ⅱ collagen-induced arthritis (CIA) was examined in DBA/1J mice in vivo. Mice were immunized intradermally twice at the 3-week interval with bovine type Ⅱ collagen(C Ⅱ). EA stimulation, begun on the 21 simultaneously with the second immunization, was applied at the acupoint equivalent to GV4 three times a week for 3 weeks. The results showed that EA delayed the onset, attenuated the severity of arthritis, and reduced the anti-collagen antibody level. Furthermore, we investigated the impact of EA on the productions of endogenous $interleukin-1{\Beta}$ (IL-1 beta) and prostaglandin E2 (PGE2), and the levels of IL-1 beta mRNA in splenocytes and synovial tissues from C Ⅱ immunized mice on the 45 and cyclooxygenase-2 (COX-2) mRNA in lipopolysaccharide (LPS)-stimulated macrophages of normal mice by using reverse transcriptase-polymerase chain reaction (RT-PCR). EA stimulation significant inhibited the concentrations of splenic endogenous IL-1 beta and serum PGE2. The expression of IL-1 beta mRNA in spleen cells was obviously down-regulated and that in synovial tissues was modestly affected by EA. COX-2 mRNA was highly expressed in cultured peritoneal macrophages when stimulated with LPS. Previous treatment with EA also reduced LPS-stimulated induction of COX-2 mRNA. These data suggest that EA has an inhibitory effect on murine CIA, and the partial mechanism of its therapeutic result may be attributed to inhibiting the productions of IL-1 beta and PGE2 by suppression the IL-1 beta and COX-2 gene activations.

• The Journal of Korean Medicine
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• v.19 no.1
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• pp.419-429
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• 1998

Poncirus trifoliata (L.) Raf (Rutaceae) fruits (PTFE) has been used for the treatment of allergic disease. IgE is normally one of the least abundant immunoglobulin (Ig) isotypes in the serum of both humans and several species of experimental animals: however a number of different stimuli can result in profound increases in IgE levels relative to other isotypes. In rodents, infection with many parasitic helminths can cause approximately 100-fold elevation in IgE within 2 wks. Immunization of mice with small amounts of protein antigens on alum also results in 10-fold to fold increase in total serum IgE, much of it specific for the immunizing antigen. In this experiment, I investigated the effect of an aqueous extract of Poncirus trifoliata (L.) Raf (Rutaceae) fruits (PTFE) on a in vivo and in vitro IgE production. PTFE dose-dependently inhibited the serum levels of IgE induced by antigens. The regulation of IgE synthesis is influenced by T cells and T cell derived factors. IL -4, a T cell-derived cytokine, has been shown to stimulate murine IgE synthesis both in vitro and in vivo. Current evidence suggests that IL-4 induces IgE synthesis in the mouse by stimulating H chain isotype switch. Lipopolysaccharide (LPS) plus IL-4 cause about l00-fold increase in IgE secretion by murine B cells. The effects of PTFE on the IL-4-dependent IgE response by mouse whole spleen cells were studied. Whole spleen cells were cultured for 7 days in the presence of LPS plus IL-4 and PTFE and the supernatants were assayed for IgE. IL-4 dependent IgE production of LPS-stimulated whole spleen cells was inhibited by PTFE. Moreover, in the present study using U266Bl human IgE-bearing B cells, I found that PTFE inhibited the production of IgE activated by LPS plus IL-4. These results indicate that PTFE have antiallergic activity by inhibition of IgE production from B cells.

• Journal of Microbiology and Biotechnology
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• v.22 no.10
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• pp.1423-1431
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• 2012

The inhibitory effect of IgY against Vibrio parahaemolyticus and Vibrio vulnificus responsible for seafood-borne diseases was investigated in this study. Water-soluble fractions (WSF) of protein containing IgYs were isolated from the egg yolk of hens initially immunized with formalin-inactivated V. parahaemolyticus or V. vulnificus. Protein, total and specific IgY contents of the WSF were determined. The inhibitory and protective effects of IgYs on the growth of V. parahaemolyticus and V. vulnificus were assayed in liquid medium and in mice. IgYs showed high affinity to their corresponding antigens with high titer from day 28 onwards. Protein contents and total IgY concentrations remained stable throughout the immunization period, whereas specific IgY concentrations increased steadily and reached a plateau at day 49. Specific IgY powder (150 mg/ml) significantly inhibited further multiplication of both V. parahaemolyticus and V. vulnificus in liquid medium as compared with the control IgY. The bacteria count in mice feces was lower in mice pretreated with specific IgYs than in those pretreated with PBS or control IgY. Higher survival of mice was observed in the experimental groups pretreated with either anti-V. parahaemolyticus (75% survival) or anti-V. vulnificus (87% survival) IgYs, compared with those in the control groups pretreated with PBS or nonspecific IgY. All mice in the control groups died within three days after bacteria inoculation; hence, the protective effect of specific IgYs against infection caused by V. parahaemolyticus and V. vulnificus was demonstrated.

• Korean Journal of Medicinal Crop Science
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• v.28 no.6
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• pp.471-480
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• 2020

Background: The roots of Cirsium japonicum var. ussuriense (RCJ) have been used as traditional medicine in Korea for hematuria and hematemesis. These extracts exert anti-oxidative and anti-inflammatory effects by scavenging for free radical and regulating the inflammatory response. However, the effect of RCJ on rheumatoid arthritis (RA) has not been elucidated. Thus, we evaluated the water extract of RCJ (WRCJ) using type II collagen-induced RA models. Methods and Results: RA was induced by immunization with type II collagen. All experimental materials were orally administered daily for three weeks. The positive control group was administered with 0.2 mg/kg methotrexate (n = 7), while the experimental group was administered with WRCJ (100 or 500 mg/kg, n = 7). Serum levels of TNF-alpha, Interleukin 6 (IL-6), and type II collagen IgG (CII) were measured using ELISA. Administration of 500 mg/kg WRCJ decreased the levels of TNF-alpha, IL-6, and CII. Moreover, WRCJ treatment diminished swelling of hind legs and infiltration of inflammatory cells in RA models' synovial membrane. Conclusions: These results indicate that WRCJ could improve RA, reduce inflammatory indicators and synovial inflammation. However, further experiments are required to determine how WRCJ can influence the signal transduction pathway in RA.

• IMMUNE NETWORK
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• v.2 no.1
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• pp.41-48
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• 2002

Background: Viral antigens presented on the cell surface in association with MHC class I molecules are recognized by CD8+ T cells. MHC restricted peptides are important in eliciting cellular immune responses. As peptide antigens have a weak immunigenicity, pH-sensitive liposomes were used for peptide delivery to induce effective cytotoxic T lymphocyte (CTL) responses. In the previous study, as the HBx peptides could induce specific CTLs in vitro, we tested whether the HLA-A2/$K^b$ transgenic mice that were immunized by HBx-derived peptides could be protected from a viral challenge. Methods: HBx-peptides encapsulated by pH-sensitive liposomes were prepared. $A2K^b$ transgenic mice were immunized i.m. on days one and seven with the indicated concentrations of liposome-encapsulated peptides. Three weeks later, mice were infected with $1{\times}10^7pfu$/head of recombinant vaccinia virus (rVV)-HBx via i.p. administration. The ovaries were extracted from the mice, and the presence of rVV-HBx in the ovaries was analyzed using human TK-143B cells. IFN-${\gamma}$ secretion by these cells was directly assessed using a peptide-pulsed target cell stimulation assay with either peptide-pulsed antigen presenting cells (APCs), concanavalin A ($2{\mu}g/ml$), or a vehicle. To generate peptide-specific CTLs, splenocytes obtained from the immunized mice were stimulated with $20{\mu}g/ml$ of each peptide and restimulated with peptide-pulsed APC four times. The cytotoxic activity of the CTLs was assessed by standard $^{51}Cr$-release assay and intracellular IFN-${\gamma}$ assay. Results: Immunization of these peptides as a mixture in pH-sensitive liposomes to transgenic mice induced a good protective effect from a viral challenge by inducing the peptide-specific CD8+ T cells. Mice immunized with $50{\mu}g/head$ were much better protected against viral challenge compared to those immunized with $5{\mu}g$/head, whereas the mice immunized with empty liposomes were not protected at all. After in vitro CTL culture by peptide stimulation, however, specific cytotoxicity was much higher in the CTLs from mice immunized with $5{\mu}g/head$ than $50{\mu}g/head$ group. Increase of the number of cells that intracellular IFN-${\gamma}$ secreting cell among CD8+ T cells showed similar result. Conclusion: Mice immunized with XEPs within pH-sensitive liposome were protected against viral challenge. The protective effect depended on the amount of antigen used during immunization. XEP-3-specific CTLs could be induced by peptide stimulation in vitro from splenocytes obtained from immunized mice. The cytotoxic effect of CTLs was measured by $^{51}Cr$-release assay and the percentage of accumulated intracellular IFN-${\gamma}$ secreting cells after in vitro restimulation was measured by flow cytometric analysis. The result of $^{51}Cr$-release cytotoxicity test was well correlated with that of the flow cytometric analysis. Viral protection was effective in immunized group of $50{\mu}g/head$, while in the in vitro restimulation, it showed more spectific response in $5{\mu}g$/head group.

• Journal of Microbiology and Biotechnology
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• v.21 no.4
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• pp.405-411
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• 2011

Avian influenza virus vaccines produced in oil-emulsified inactivated form with antigen content of at least 160 hemagglutinin units (HAU) induced immunity in birds. However, in addition to enhancing the effect of the adjuvant(s), other additional supplemented biological compounds included in inactivated vaccines could produce higher levels of antibody. We examined in chickens, Vietnamese ducks, and muscovy ducks the adjuvant effect of Sophy ${\beta}$-glucan (SBG), a ${\beta}$-1,3-1,6 glucan produced by the black yeast Aureobasidium pollulans strain AF0-202, when administered with an avian influenza H5 subtype vaccine. In Experiment 1, 40 chickens (ISA Brown hybrid), allocated to four groups of ten each, were immunized with Oil-H5N1(VN), Oil-H5N1(CN), Oil-H5N2(CN), and saline (control group), respectively. In Experiment 2, chickens (ISA Brown hybrid), muscovy ducks (French hybrid), and Vietnamese ducks (indigenous Vietnamese) were used to further assess the effect of SBG on immunogenicity of the Oil-H5N1(VN) Vietnamese vaccine. ELISA and hemagglutination inhibition (HI) assays were used to assess the antibody response. The H5 subtype vaccines initiated significantly higher immune responses in the animals dosed with SBG, with 1.0-1.5 $log_2$ higher HI titers and 10-20% ELISA seroconversion, compared with those not dosed with ${\beta}$-glucan. Notably, some of the animals dosed with SBG induced HI titers higher than 9.0 $log_2$ following boosting immunization. Taken together, our serial studies indicated that SBG is a potential effector, such as enhancing the immune response to the H5 vaccines tested.

• Korean Journal of Animal Reproduction
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• v.11 no.1
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• pp.63-72
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• 1987

Effects of sperm and seminal plasma isoant ibodies upon the rate of superovulation and ferlil ization were slud ied in both normal and immunized rabbits. The results obstained were summarized as follows: 1. On examin.ltion of the superovulation in the immunized animals, the average number of ovulation points was 22.1 and 25.3 for spermtreated animals and for seminal plasma-treated animals, respec tively. As compared to the con¬trol group of 41.0 in number, the immunized showed statisfical significance in ovulation (P<0.05). 2. In ovary weight and follicle's size there were no significant differences among the three groups, whereas sperm-and seminal plasma¬treated groups had an average rate of fertiliza¬tion of 62.X% and 5X .0%, respectively, in re¬markable contrast to the control hTfOUP of 91.4'1'0 (P<0.05). 3. When the animals were inseminated with a mixture of semen plus sperm or seminal plasma antisera, a sharp reduction of fertilization was observed with 5.6% and 16.0% as compared to the control group (P<0.05). Consequently, immunization with either sperm or seminal plasma had a substantial effect on fertilization.

• Journal of Acupuncture Research
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• v.28 no.1
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• pp.37-44
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• 2011

배경 및 목적 : 봉독약침요법(bee venom pharmacopuncture, BVP)은 rheumatoid arthritis(RA)의 치료에 활용되고 있으나, RA로 인한 염증성 통증에 대한 봉독약침의 진통효과와 specific mechanism은 아직까지 명확하게 밝혀지지 않았다. 이에 본 연구에서는 RA animal model로서 collagen-induced arthritis(CIA) rat model에서 봉독약침의 a1-adrenergic, 5HT-3 그리고 muscarinic cholinergic mechanism을 확인하고자 한다. 방법 : CIA를 유도하기 위하여 male Sprague-Dawley rat에 freund's incomplete adjuvant에 유화(乳化)시킨 bovine type II collagen을 주입하고 14일 후 booster injection 시행하였다. 진통효과는 tail flick latency (TFL)로 평가하였다. 결과 : 관절염의 유도 이후 염증성 통증 역치는 시간이 지나면서 낮아지며, 5주 이후로는 통증 역치에 큰 변화가 없이 유지되었다. 첫 번째 immunization으로부터 5주 경과 후 족삼리($ST_{36}$)에 봉독약침처치(0.25 mg/ kg)를 시행하여 유의한 진통효과를 관찰하였다. 또한 봉독약침의 진통효과는 ondansetron(5HT-3 receptor antagonist, 0.5mg/kg, i.p.), atropine(muscarinic cholinergic receptor antagonist, 1mg/kg, i.p.)의 전처치에 의하여 억제되었으나, prazosin(a1-adrenergic receptor antagonist, 1mg/kg, i.p.)의 전처치에 의해서는 억제되지 않았다. 결론 : 봉독약침은 CIA로 인한 염증성 통증에 유의한 진통효과를 나타내며 그 analgesic mechanism은 5HT-3와 muscarinic cholinergic receptor에 의하여 매개되며 a1-adrenergic receptor에 의하여 매개되지는 않았다.

• Journal of Periodontal and Implant Science
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• v.30 no.3
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• pp.675-687
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• 2000

Multiple periodontal pathogens sequentially colonize the subgingival niche during the conversion from gingivitis to destructive periodontal disease. An animal model of sequential immunization with key periodontal pathogens has been developed to determine whether T and B lymppocyte effector functions are skewed and fail to protect the host from pathogenic challenge. The present study was performed to evaluate immunomodulatory effect of exposure to Fusobacterium nucleatum(F. nucleatum) prior to Porphyromonas gingivalis(P. gingi - valis). Group 1(control) mice were immunized with phosphate-buffered saline, Group 2 were immunized with F. nucleatum prior to P. gingivalis, while Group 3 were immunized P. gingivalis alone. All the T cell clones derived from Group 2 demonstrated type 2 helper T cell clone(Th2 subsets), while those from Group 3 mice demonstrated Th1 subsets. Exposure of mice to F . nucleatum prior to P. gingivalis interfered with opsonophagocytosis function of sera against P. gingivalis. In adoptive T cell transfer experiments, in vivo protective capacity type 2 helper T cell clones(Th2) from Group 2 was significantly lower than type 1 helper T cell clones(Th1) from Group 3 against the lethal dose infection of P. gingivalis. Western blot analysis indicated the different pattern of recognition of P .gingivalis fimbrial proteins between sera from Group 2 and Group 3. In conclusion, these study suggest that colonization of the subgingival niche by F .nucleatum prior to the periodontal pathogen, P. gingivalis, modulates the host immune responses to P. gingivalis at humoral, cellular and molecular levels.