Kim, Seong-Geun;Ran, Gui Shao;Kwon, Hyuk-Chan;Hwang, Tae-Ho
Journal of Life Science
/
v.26
no.1
/
pp.23-33
/
2016
Pexa-Vec (JX-594) is a specific cancer-targeted oncolytic and immunotherapeutic vaccinia virus. The purpose of this study was to develop methods to maximize the stability of Pexa-Vec. In short-term instability testing, viral activity was rapidly decreased both at 4℃ and at room temperature (RT), but it was completely restored after sonication followed by vortex. Long-term stability testing of Pexa-Vec in the following liquid formulations was performed: (A) 30 mM Tris/pH 7.6, (B) 30 mM Tris/pH 8.6, (C) 30 mM Tris/pH 7.6, 150 mM NaCl, 15% sucrose, (D) 30 mM Tris/pH 7.6, 15% sucrose, and (E) 30 mM Tris/pH 8.6, 15% sucrose. Viral activity decreased less than 2 log10 at 4℃, and RT was observed in 3 days in B, while viral activity was not decreased even after 4–8 weeks at 4℃ and at 1 week in RT in A, suggesting that neutral pH may be essential to maintain virus stability. The addition of 15% sucrose into A (D) significantly increased viral stability at −20℃, 4℃, or RT, and it was also observed at pH 8.6 (E). The addition of 150 mM NaCl into D (C) significantly increased viral stability in addition to the sucrose effect at 4℃ or RT. Accordingly, the viral activity in formulation C was maintained for 1.5 years at 4℃, and for 1-2 weeks in RT. In conclusion, we propose that formulation C can provide the most adequate condition for the proper storage of vaccinia oncolytic virus.
Kim, Do-Soon;Park, Jueng-Eun;Seo, Kwon-Il;Ko, Sung-Ryong;Lee, Jong-Won;Do, Jae-Ho;Yee, Sung-Tae
Journal of Ginseng Research
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v.30
no.3
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pp.117-127
/
2006
Ginseng is a medicinal herb widely used in Asian countries. Dendritic cells(DCs) play a pivotal role in the initiation of T cell-mediated immune responses, making them an attractive cellular adjuvant for use in cancer vaccines. In this study, we examined the effects of Red-ginseng(water extract, edible and fermented ethyl alcohol extract, crude saponin) on the DCs phenotypic and functional maturation. Immature DCs were cultured in the presence of GM-CSF and IL-4, and the generated immature DCs were stimulated by water extract, edible and fermented ethyl alcohol extract, crude saponin and LPS, respectively, for 24hours. The expression of surface co-stimulatory molecules, including MHC(major histocompatibility complex) class II, CD40, CD80 and CD86, was increased on DCs that were stimulated with crude saponin, but antigen-uptake capacity was decreased. The antigen-presenting capacity of Red-ginseng extracts-treated DCs as analyzed by allogeneic T cells proliferation and IL-2, $IFN-{\gamma}$ production was increased. Furthermore, $CD4^+$ and $CD8^+$ syngeneic T cell(OVA-specific) proliferation and $IFN-{\gamma}$ production was significantly increased. However, $CD4^+$ syngeneic T cell secreted higher levels of IL-2 in responding but not $CD8^+$ syngeneic T cell. These results indicate the immunomodulatory properties of Red-ginseng extracts, which might be therapeutically useful in the control of cancers and immunodeficient diseases through the up-regulation of DCs maturation.
Journal of the Korean Society of Food Science and Nutrition
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v.32
no.8
/
pp.1357-1363
/
2003
This study was conducted to investigate the effect of chicken egg containing IgY against H. pylori in patients with gastritis. Sixty three H. pylori-infected volunteers (20∼43 year, Male Female=49 : 14) were randomized into four groups which were treated with one chicken egg containing IgY b.i.d. (IgY group; n=17) or omeprazole 20 mg b.i.d., amoxicillin 1.0 g b.i.d. and clarithromycin 500 mg b.i.d. (OAC group; n=17) or omeprazole 20 mg b.i.d., amoxicillin 1.0 g b.i.d., clarithromycin 500 mg b.i.d. and one chicken egg containing IgY b.i.d. (OAC with IgY group; n=16) for 2 weeks or lyophilized IgY 1 g b.i.d (lyophilized IgY group) for 1 month. $\Delta$$^{13}$$CO_2$ before and after treatment, the eradication rate of H. pylori and histologic change including H. pylori density, acute and chronic inflammation activity, intestinal metaplasia and glandular atrophy by updated sydney system were evaluated. Eradication rate of OAC with IgY group (94%) was higher than IgY group (0%), lyophilized IgY group (0%) and OAC group (88%). $\Delta$$^{13}$$CO_2$at 2 weeks after treatment in one patient of IgY group was decreased. But that was not changed in the other patients. $\Delta$$^{13}$$CO_2$ at 1 week after treatment in 15 patients of OAC with IgY group was significantly lower than pretreatment level (p<0.05), and $\Delta$$^{13}$$CO_2$ at 1 week and 2 week after treatment was decreased in the other patient. Acute inflammation activity at antrum was significantly decreased after treatment in IgY and lyophilized IgY group (p<0.01), H. pylori density at antrum was significantly decreased after treatment in IgY and lyophilized IgY group (p<0.05). Chronic inflammation activity at body was decreased after treatment in lyophilized IgY group. Intestinal metaplasia and glandular atrophy at antrum and body were not changed after treatment in IgY group. Mild intestinal metaplasia in one patient of lyophilized IgY group changed to normal after 1 month treatment. Gandular atrophy at antrum and body were not changed after treatment in lyophilized IgY group.
Eugenol (4-allyl-2-methoxyphenol) is a natural phenolic constituent extensively used in dentistry as a component of zinc oxide eugenol cement and is applied to the mouth environment. Chios gum mastic (CGM) is a resinous exudate obtained from the stem and the main leaves of Pistacia lenticulus tree native to Mediterranean areas. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with a natural product, CGM and natural phenolic compound, eugenol on SCC25 human tongue squamous cell carcinoma cell line. To investigate whether the co-treatment with eugenol and CGM compared to each single treatment efficiently reduces the viability of SCC25 cells, MTT assay was conducted. Induction and augmentation of apoptosis were confirmed by Hoechst staining, TUNEL staining and DNA hypoploidy. Westen blot analysis and immunofluorescent staining were performed to study the alterations of the expression level and the translocation of apoptosis-related proteins in co-treatment. In this study, co-treatment of with eugenol and CGM on SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensations, DNA fragmentation, the increase and decrease of Bax and Bcl-2, decrease of DNA content, the release of cytochrome c into cytosol, translocation of AIF and DFF40 (CAD) onto nuclei, and activation of caspase-3, caspase-6 caspase-7, caspase-9, PARP, Lamin A/C and DFF45 (ICAD) whereas each single treated SCC25 cells did not show or very slightly these patterns. Although the single treatment of 40 ${\mu}g$/ml CGM and 0.5 mM eugenol for 24 h did not induce apoptosis, the co-treatment of these reagents prominently induced apoptosis. Therefore our data provide the possibility that combination therapy with CGM and eugenol could be considered as a novel therapeutic strategy for human oral squamous cell carcinoma.
Kim, Jae-Yeol;Lee, Jae-Cheol;Kang, Min-Jong;Park, Jae-Seok;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Lee, Jae-Ho
Tuberculosis and Respiratory Diseases
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v.42
no.5
/
pp.703-712
/
1995
Background: Peripheral blood monocytes are important immune effector cells that play a fundamental role in cellular immunity. In addition to their antigen-presenting and phagocytic activities, monocytes/macrophage produce a vast array of regulatory and chemotactic cytokines. Interleukin-8(IL-8), a potent neutrophil-activating and chemotactic peptide, is produced in large quantities by mononuclear phagocytes and may be an important mediator of local and systemic inflammation. Overexpression by IL-8 of such inflammation may be an important step of tissue injury frequently seen in inflammatory reaction. So it could be hypothesized that the agents which block the production of IL-8 can decrease the inflammatory reaction and tissue injury. To evaluate this, we described the effect of Dexamethasone, $PGE_2$, Indomethacin and Interferon-$\gamma$(IFN-$\gamma$) on IL-8 mRNA and protein expression from LPS-stimulated human peripheral blood monocytes(PBMC). Method: PBMC was isolated from healthy volunteers. To evaluate the effect of Dexamethasone, $PGE_2$ & Indomethacin, these drug were treated for 1 hour before and after LPS stimulation and IFN-$\gamma$ was only treated I hour before the LPS stimulation. Northern blot analysis for IL-8 mRNA and ELISA for immunoreactive IL-8 protein in culture supernatant were performed. We repeated above experiment three times for Northern blot analysis and two times for ELISA and got the same result. Results: 1) Pre- and post-treatment of Dexamethasone suppressed both the LPS stimulated IL-8 mRNA expression and IL-8 protein release in PBMC. 2) IFN-$\gamma$ pre-treatment suppressed the IL-8 mRNA expression and IL-8 protein release in unstimulated cells. 3) In LPS stimulated cells, IFN-$\gamma$ suppressed the IL-8 mRNA expression but IL-8 protein release suppression was not observed. 4) $PGE_2$ and Indomethacin exert no effect on the LPS-stimulated IL-8 mRNA and protein expression in concentration used in this experiment ($PGE_2;10^{-6}M$, Indomethacin; $10{\mu}M$). Conclusion: One of the mechanism of antiinflammatory action of Dexamethasone can be explained by the suppressing effect of IL-8 production in some extent and by this antiinflammatory effect, dexamethasone can be used to suppress local and systemic inflammation mediated by IL-8.
Son, Ji Yoon;Park, Young W.;Renchinkhand, Gereltuya;Han, Jung Pil;Bum, Jin Woo;Paik, Seung-Hee;Lee, Jo Yoon;Nam, Myoung Soo
Journal of Life Science
/
v.26
no.6
/
pp.689-697
/
2016
Kefir is an acidic-alcoholic fermented milk product originating from the Caucasian mountains. Kefir has long been known for its probiotic health benefits, including its immunomodulatory effects. The objectives of this study were to investigate the properties of a fermented whey product and to examine the effects of kefir grains on the in vitro immune-modulation of human mast cell-1 (HMC-1). The results showed that the whey fermented by kefir grains contained the maximum lactic acid bacteria and yeast for 16 hr by 1.83×108 and 6.5×105 CFU/ml, respectively, and lactose and whey proteins were partially hydrolyzed. The experimental whey fermented by kefir grains exhibited an in vitro anti-inflammatory effect on the HMC-1 line for 8, 16, and 24 hr, and this effect induced the expression of interleukin (IL)-4 as a pro-inflammatory cytokine, but not for 48 hr by RT-PCR in HMC-1 cells. In addition, the same phenomenon was observed for the expression of IL-8 as a pro-inflammatory cytokine by the kefir-fermented whey during the same periods of 8-48 hr under the same conditions. These cytokines resulted in the production of IL-4 at 20-25 ng in HMC-1 cells for 8, 16, and 24 hr, whereas 5 ng was produced for 48 hr by the fermented whey. In contrast, IL-8 was produced at 15-20 ng in HMC-1 cells during 4, 8, 16, and 24 hr, while 7 ng was produced at 48 hr. It was concluded that the whey fermented by kefir grains possesses a potential anti-inflammatory function, which could be used for an industrial application as an ingredient of functional foods and pharmaceutical products.
Park, Se-Il;Moon, Young-Mi;Jeong, Jae-Ho;Jang, Kwang-Ho;Ahn, Myun-Hwan
Journal of Veterinary Clinics
/
v.28
no.5
/
pp.486-496
/
2011
A special mesenchymal tissue layer called perichondrium has a chondrogenic capacity and is a candidate tissue for engineering of cartilage. To overcome limited potential for chondrocyte proliferation and re-absorption, we studied a method of cartilage tissue engineering comprising chondrocyte-hydrogel pluronic complex (CPC) and cultured perichondrial cell sheet (cPCs) which entirely cover CPC. For effective cartilage regeneration, cell-sheet engineering technique of high-density culture was used for fabrication of cPCs. Hydrogel pluronic as a biomimetic cell carrier used for stable and maintains the chondrocytes. The human cPCs was cultured as a single layer and entirely covered CPC. The tissue engineered constructs were implanted into the dorsal subcutaneous tissue pocket on nude mice (n = 6). CPC without cPCs were used as a controls (N = 6). Engineered cartilage specimens were harvested at 12 weeks after implantation and evaluated with gross morphology and histological examination. Biological analysis was also performed for glycosaminoglycan (GAG) and type II collagen. Indeed, we performed additional in vivo studies of cartilage regeneration using canine large fullthickness chondrial defect model. The dogs were allocated to the experimental groups as treated chondrocyte sheets with perichondrial cell sheet group (n = 4), and chondrocyte sheets only group (n = 4). The histological and biochemical studies performed 12 weeks later as same manners as nude mouse but additional immunofluorescence study. Grossly, the size of cartilage specimen of cPCs covered group was larger than that of the control. On histological examination, the specimen of cPCs covered group showed typical characteristics of cartilage tissue. The contents of GAG and type II collagen were higher in cPCs covered group than that of the control. These studies demonstrated the potential of such CPC/cPCs constructs to support chondrogenesis in vivo. In conclusion, the method of cartilage tissue engineering using cPCs supposed to be an effective method with higher cartilage tissue gain. We suggest a new method of cartilage tissue engineering using cultured perichondrial cell sheet as a promising strategy for cartilage tissue reconstruction.
This research was performed to determine the proximate compositions, mineral contents, alginic acid, antioxidative activities and amino acids of sea tangles collected from Gijang and Wando area. Crude protein and ash contents were higher in Gijang sea tangle, whereas carbohydrate and moisture were higher in Wando in general. Mineral contents of Gijang sea tangle were higher than Wando. Especially, Na and K was the most abundant in both Gijang and Wando sea tangles. Alginic acid content was almost similar in both sea tangles. The major free amino acids were glutamic acid, aspartic acid, alanine, proline and hydroxyproline in both Gijang and Wando sea tangles. Antioxidative activity of methanol extract of sea tangle was measured by using DPPH radical scavenging and SOD-like activity. DPPH radical scavenging and SOD-like activity were about 17% ($40\;{\mu}g/ml$) and 7% ($5\;{\mu}g/ml$) higher, respectively, in Wando sea tangle. When stimulate the macrophages RAW264.7 cells with lipopolysaccharide (LPS), inhibition of NO synthesis of the methanol extract was 11% higher in Wando sea tangle comparing with Gijang samples.
With the incidence of antibiotic resistant bacteria there is increasing interest in natural products such as herb extract and probiotics to control antibiotic resistant bacteria. This study was focused on the determination of antimicrobial activity of Opuntia ficus-indica var. saboten against Salmonella enetrica serovar Enteritidis (S. enterifidis), S. enterica serovar Typhimurium (S. Typhimurium) DT 104 and Escherichia coli 0157:H7. Though bactericidal effect of 0. ficus-indica var. saboten was not observed, it had significant inhibitory activity against Salmonella spp. and E. coli O157:H7 on the Moulter Hinton agar containing its solution dissolved in deionized water. To investigate the antimicrobial activity in vivo, mice were challenged with 5. Typhimurium DT104 (3.7$\times$108 cfu/mouse) after pre-feeding 0. ficus-indica var. saboten solution. The fecal shedding of S. Typhimurium DT104 was more dramatically decreased and not detectable in feces and intestines 3 days after challenge in mice fed with 0. ficus-indica var. saboten. Antibody responses of the intestinal IgA were also significantly increased in mice fed with 0. ficus-indica var. saboten. These findings suggest that Opuntia ficus-indica var. saboten decreased the shedding of S. Typhimurium DT104 in vitro and also in the gastrointestinal tract in mice. In addition, administration of the product might enhance the mucosal immune response against S. Typhimurium DT 104. In conclusion, Opuntia ficus-indica var. saboten might be useful to control antibiotic resistant bacteria in vivo and in vitro.
Shin, Ji Eun;Lee, Kyungmin;Kim, Ji-Hee;Madhi, Iskander;Kim, YoungHee
Journal of Life Science
/
v.29
no.4
/
pp.402-409
/
2019
Korean red ginseng made from steaming and drying fresh ginseng has long been used as a traditional herbal medicine due to its effects on the immune, endocrine, and central nerve systems and its anti-inflammatory activity. In this study, we investigated the molecular mechanism responsible for the anti-inflammatory effects of a formulated Korean red ginseng extract (RGE) in response to lipoteichoic acid (LTA), a cell wall component of gram-positive bacteria. RGE inhibited LTA-induced nitric oxide (NO) secretion and inducible nitric oxide synthase (iNOS) expression in BV-2 microglial cells, without affecting cell viability. RGE also inhibited nuclear translocation of nuclear factor kappa B ($NF-{\kappa}B$) p65 and degradation of $I{\kappa}B-{\alpha}$. In addition, RGE increased the expression of heme oxygenase-1 (HO-1) in a dose-dependent manner, and the inhibitory effect of RGE on iNOS expression was abrogated by small interfering RNA-mediated knockdown of HO-1. Moreover, RGE induced nuclear translocation of nuclear factor E2-related factor 2 (Nrf2), a transcription factor that regulates HO-1 expression. Furthermore, the phosphoinositide-3-kinase (PI-3K) inhibitor and mitogen-activated protein kinase (MAPK) inhibitors suppressed RGE-mediated expression of HO-1, and RGE enhanced the phosphorylation of Akt, extracellular signal-regulated kinases (ERKs), p38, and c-JUN N-terminal kinases (JNKs). These results suggested that RGE suppressed the production of NO, a proinflammatory mediator, by inducing HO-1 expression via PI-3K/Akt- and MAPK-dependent signaling in LTA-stimulated microglia. The findings indicate that RGE could be used for the treatment of neuroinflammation induced by grampositive bacteria and that it may have therapeutic potential for various neuroinflammation-associated disorders.
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