• Journal of the Korean Wood Science and Technology
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• v.24 no.1
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• pp.68-74
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• 1996

Present work was undertaken to investigate the origin of milled wood lignin(MWL) in the wood cell wall using immunocytochemical techniques, which can provide the information on the localization of specific antigens(MWL in the present study) to be examined. Spruce MWL dissolved in DMSO and emulsified with Freund adjuvant was injected directly into the mouse spleen. The animals were boostered at two-week intervals after the initial immunization. Blood samples were purified in standard procedures. The characteristics of antibodies against MWL were tested by indirect ELISA. Visualization of MWL was carried out using conventional indirect immunogold-labelling methods on the ultrathin sections of spruce wood. Immuno-TEM observations showed that the immunogold probes were selectively attached to secondary cell walls of spruce wood. The most intense labelling was frequently observed in the S2 layer. In contrast, gold labelling in the lignin-rich regions, such as middle lamella and cell corner was not found. The immuno-TEM provides an indication that spruce MWL originates from the S2 layer.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.543-547
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• 2002

DNA vaccines use eukaryote expression vectors to produce immunizing proteins in the vaccinated host and it represents a novel approach to vaccine and immuno-therapeutic development. We constructed a 2.9 kb compact plasmid vector (pVAC) which contains CMV promoter, polycloning site, BGH poly A terminator, ampicillin resistance gene and PBR322 origin. Enriched unmathlyated CpG motifs have introduced into pVAC-ISS1 and pVAC-ISS2 which are derived from pVAC for enhancing Thl responses. These plasmid DNAs rapidly induced interleukin 6 secretion in vivo. It is expected that these vectors will contribute to the DNA inoculation against infectious disease and various cancers without adjuvant.

• IMMUNE NETWORK
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• v.3 no.2
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• pp.156-163
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• 2003

Background: $\beta$-1,3-glucans are well known to enhance the immune reactions, resulting in antitumor, antibacterial, antiviral, anticoagulatory and wound healing activities. $\beta$-1, 3-glucans have various activities depending on molecular weight, degree of branching, conformation, water-solubility and intermolecular association. However, the $\beta$-1,3-glucan linked backbone structure is essential and $\beta$-D-glucopyranosyl units are required for immuno-potentiating activities. Result: In this study, we tested the immunophamacological activities of soluble $\beta$-1,3-glucans and confirmed the following activities: (1) $IFN-{\gamma}$ production in PBMCs in the presence or the absence of PHA, LPS, or IL-18; (2) induction of various cytokines in the spleen and thymus; (3) adjuvant effect on the antibody production; (4) nitrogen oxide synthesis in macrophages; (5) the cytotoxic and antitumor effects on cell lines and ICR mice. Conclusion: These results strongly suggested that $\beta$-1,3-glucans possessed various immuno-pharmacological activities.

• YAKHAK HOEJI
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• v.56 no.1
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• pp.20-25
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• 2012

In Northeast Asia, Paeoniae Radix has been used in treatments of inflammation-causing diseases such as arthritis for many centuries. Paeoniflorin, one of the principle bioactive monoterpene glucosides from the paeony root, is reported to be mostly responsible for the effectiveness of the treatments. However, the anti-inflammatory effect of a monoterpene, paeoniflorigenone (PFG) which partially has the moiety of paeoniflorin minus a glucose structure is unknown. Thus, the aim of this work was to investigate anti-inflammatory activity of PFG. For the investigation, PFG activity on the NO (nitric oxide) production from LPS-stimulated macrophages, and the anti-inflammatory effect was tested in the animal model of septic arthritis caused by Candida albicans, a major etiological agent for septic arthritis. For induction of the arthritis, mice were administered with an emulsion of C. albicans cell wall (CACW) mixed with Complete Freund's Adjuvant (CFA) via footpad-injection (Day 0); PFG at a dose of 0.5 or 1 mg/mouse (25 or 50 mg/kg of body-weight) was given to the animals on Day 3, 6, and 9; footpads were scored for arthritis. Moreover, the PFG effect on proliferation of T-lymphocyte that causes aggravation of arthritis was additionally tested. Data resulting from those tests showed that PFG inhibited the NO production from the stimulated macrophage in a dose dependent manner (P<0.05), indicating that PFG is an anti-inflammatory. To confirm the in-vitro results, anti-inflammatory activity of PFG was determined against C. albicans-caused septic arthritis. Data showed that PFG-treatment reduced footpad-swelling which indicates that PFG has anti-arthritic effect (P<0.05), which is therapeutic. The anti-arthritic effect appeared to be mediated by PFG suppression of T-cell proliferation. Ultimately, PFG, a monoterpene component, has anti-inflammatory activity analogous to paeoniflorin. The anti-inflammatory activity treats the septic arthritis due to a pathogenic fungus C. albicans.

• Korean Journal of Veterinary Research
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• v.40 no.3
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• pp.575-585
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• 2000

This study was performed to observe the influence of host immune response on the chemotherapy of mice experimentally infected with Fasciola hepatica. Following immunosuppression with prednisolone or immunoenhancement with Freund's complete adjuvant(FCA), mice were experimentally infected with 3 Fasciola hepatica metacercariae and treated with closantel at 1 week post infection. In the group of mice infected with metacercariae alone, 2 mice of 10 were dead at 10 weeks post infection(20% mortality), and adult flukes were recovered from the liver and the peritoneal cavity of the remaining 8 mice(100% infectivity). In the group of mice treated with prednisolone and infected with metacercariae, 8 of 10 mice died before euthanasia with a mean time of death earlier than the control group (p<0.05). In the group of immunosuppressed mice infected with metacercariae and treated with closantel 20mg/kg, 4 of 10 mice died before sacrifice. In the group of mice infected and treated with closantel 20mg/kg, mortality and infectivity was 10% and 30%, respectively. Similar results were observed in mice infected and treated with closantel 5mg/kg which resulted in 10% and 50% mortality and infectivity, respectively. These results indicated that the efficacy of closantel treatment was decreased in immunosuppressed mice, while the pathogenicity was increased. In immunoenhanced mice infected with metacercariae, on the other hand, the efficacy of chemotherapy with both 5mg/kg or 20mg/kg closantel resulted in only 10% infectivity. The results shown in this study strongly suggest that a close interaction between chemotherapy against F hepatica with closantel and the host immune system exists. Considering that fascioliasis is a zoonosis, treatment regimen against the infection to immunosuppressed patients may require a concurrent prescription of an appro-priate immuno-enhancing adjuvant.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.1
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• pp.29-33
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• 2005

A DNA vaccine methodology using eukaryote expression vectors to produce immunizing proteins in the vaccinated hosts is a novel approach to the development of vaccine and immuno-therapeutics, and it has achieved considerable success over several infectious diseases and various cancers. To further enhance its efficiency, attempts were made to develop novel plasmid vectors containing multiple immunostimulatory CpG motifs, for rapid and strong immune response. First, a 2.9 kb compact plasmid vector (pVAC), containing CMV promoter, polycloning site, BGH poly(A) terminator, ampicillin resistance gene and pBR322 origin was constructed. A pVAC-hEPO was also constructed, which contained a human erythropoietin gene, for evaluating the transfection efficiency of naked plasmid DNA both in vitro and in vivo. To examine the adjuvant effect of multi-CpG motifs on naked plasmid DNA, 22 and 44 enriched and unmethylated CpG motifs were introduced into pVAC to generate pVAC-ISS1 and pVAC-ISS2, respectively. $100{\mu}g$ of pSecTagB, pVAC, pVAC-ISS1 or pVAC-ISS2 were each injected intramuscularly into the tibilias anterior muscle of Balb/c mice. The level of interleukin-6 induced in the mice injected with pVAC-ISS1 and pVAC-ISS2 were significantly elevated after 12 hours, which were almost 2 and 2.5 times higher than that in the mice injected with pSecTagB, respectively. These results suggest that DNA vaccine plasmids with enriched CpG motifs can induce rapid secretion of interleukin-6 by lymphocytes. In conclusion, these vectors can contribute to the development of adjuvant-free DNA vaccinations against infectious diseases and various cancers.

• Parasites, Hosts and Diseases
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• v.30 no.2
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• pp.133-140
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• 1992

Theileria sergenti were isolated from infected erythrocytes by hypotonic Iysis, and soluble meroBoite antigens were purified by sonication and differential centrifugation. The preparation contained 29, 34, 35 and 105 kD immuno-dominant polypeptides. The soluble antigens (0.5 mg/ml) were prepared and fortified with Freund's adjuvant. Five month old naive Korean calves were subcutaneously inoculated with the preparation and a booster dose was administered 4 weeks later Nine weeks after the booster dose, vaccinates and controls were challenged with a homologous stabilate (5.6×106 RBC/dose, 40% Parasitemia). All animals were monitored for hematocrit, total erythrocyte count, parasitemia and for the specific antibody by Western immuno- blot (WB) and indirect immuno-auorescent antibody(IFA) test. By 18 weeks after vaccination (6 weeks after the challenge), vaccinated cattle had an average IFA titer of 1 : 10,240 compared with 1 : 1,280 of the controls. The vaccinates showed ne91igib1e change in hematocrit and total RBC count whereas control animals showed significant (P<0.05) hematological chanties and associated anemia. After vaccination and challenge, the antibody responses demonstrated that vaccination had induced significant production of antibody to the 29 and 35 kD polypeptides. The latter polypeptide was much more strongly recognized by the vaccinated animals, and thus it may be a potential candidate for the vaccine.

• The Korean Journal of Food And Nutrition
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• v.29 no.3
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• pp.431-437
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• 2016

Macrophages play a pivotal role in the innate and adaptive immune systems. This study investigated the immuno-modulatory activities of polysaccharides separated from Chrysanthemum zawadskii var. latilobum (CZPS) in macrophages. Polysaccharides from Chrysanthemum zawadskii var. latilobum were extracted by the ethanol precipitation method. RAW 264.7 mouse macrophage cell line was treated with CZPS (4 to $128{\mu}g/mL$), and there was no cytotoxicity at a dose below $32{\mu}g/mL$. The levels of nitric oxide (NO) and pro-inflammatory cytokines (tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-6, IL-$1{\beta}$) production in the CZPS treated group ($32{\mu}g/mL$) were $6.5{\pm}0.12{\mu}m$ (NO), $1252.8{\pm}79.85$ (TNF-${\alpha}$), $305.4{\pm}29.41$ (IL-6), and $683.3{\pm}59.71$ (IL-$1{\beta}$), respectively, and they were significantly increased when compared to the control group; $2.2{\pm}0.03{\mu}m$ (NO), $452.3{\pm}38.34$ (TNF-${\alpha}$), $31.7{\pm}5.75$ (IL-6), and $184.1{\pm}11.52$ (IL-$1{\beta}$). Additionally, protein expression of inducible nitric oxide synthase (iNOS) and phosphorylation of MAPKs and NF-${\kappa}B$ expression were significantly increased upon CZPS treatment. Therefore, these results indicated that polysaccharides separated from Chrysanthemum zawadskii var. latilobum (CZPS) may have a potential immunomodulatory activity in macrophages through MAPKs and NF-${\kappa}B$ signaling, and this information is useful for the development of immune enhancing adjuvant materials using a natural ingredient.

• The Korean Journal of Food And Nutrition
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• v.17 no.1
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• pp.47-52
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• 2004

In this study, I attempted to develope the expression and purification system of human mammaglobin proteins in Escherichia coli and to produce anti-human mammaglobin rabbit antibody for the detection of human mammaglobin protein in the peripheral blood of breast cancer patients. Human mammaglobin gene was cloned and sequenced from m-RNAs purified from donated breast cancer tissues using RT-PCR. The cloned gene was inserted into pET30, pET22, and pET32 plasmid. The cloned gene in pET30 yields insoluble proteins which was difficult to purify from the cells extracts. The mammaglobin gene in pET32 was strongly expressed soluble proteins which were isolated using Ni-NTA affinity chromagraphy and DEAE-ion exchange chromatography, followed by enterokinase digestion of the purified proteins. The isolated proteins had enough purity to use as a antigen for the production of anti-mammaglobin antibody in rabbits. The polyclonal antibody produced against the isolated mammaglobin showed a specificity to mammaglobin after Westernblot immuno assay. In conclusion, the isolated mammaglobin protein and the anti-mammaglobin rabbit antibody may be used for diagnosis of breast cancer as well as development of anti-breast cancer drug.

• BMB Reports
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• v.39 no.6
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• pp.696-702
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• 2006

Recently three proteins, playing central roles in the bidirectional transport of urate in renal proximal tubules, were identified: two members of the organic anion transporter (OAT) family, OAT1 and OAT3, and a protein that designated renal urate-anion exchanger (URAT1). Antibodies against these transporters are very important for investigating their expressions and functions. With the cytokine gene as a molecular adjuvant, genetic immunization-based antibody production offers several advantages including high specificity and high recognition to the native protein compared with current methods. We fused high antigenicity fragments of the three transporters to the plasmids pBQAP-TT containing T-cell epitopes and flanking regions from tetanus toxin, respectively. Gene gun immunization with these recombinant plasmids and two other adjuvant plasmids, which express granulocyte/macrophage colony-stimulating factor and FMS-like tyrosine kinase 3 ligand, induced high level immunoglobulin G antibodies, respectively. The native corresponding proteins of URAT1, OAT1 and OAT3, in human kidney can be recognized by their specific antibodies, respectively, with Western blot analysis and immunohistochemistry. Besides, URAT1 expression in Xenopus oocytes can also be recognized by its corresponding antibody with immuno-fluorescence. The successful production of the antibodies has provided an important tool for the study of UA transporters.