• Journal of Microbiology and Biotechnology
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• 제28권6호
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• pp.1022-1029
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• 2018

Tuberculosis (TB) remains a serious health issue around the word. Adenovirus (Ad)-based vaccine and modified vaccinia virus Ankara (MVA)-based vaccine have emerged as two of the most promising immunization candidates over the past few years. However, the performance of the homologous and heterologous prime-boost immunization regimens of these two viral vector-based vaccines remains unclear. In the present study, we constructed recombinant Ad and MVA expressing an Ag85B-TB10.4 fusion protein (AdH4 and MVAH4) and evaluated the impact of their different immunization regimens on the humoral and cellular immune responses. We found that the viral vector-based vaccines could generate significantly higher levels of antigen-specific antibodies, $IFN-{\gamma}$-producing splenocytes, $CD69^+CD8^+$ T cells, and $IFN-{\gamma}$ secretion when compared with bacillus Calmette-$Gu{\acute{e}}rin$ (BCG) in a mouse model. AdH4-containing immunization regimens (AdH4-AdH4, AdH4-MVAH4, and MVAH4-AdH4) induced significantly stronger antibody responses, much more $IFN-{\gamma}$-producing splenocytes and $CD69^+CD8^+$ T cells, and higher levels of $IFN-{\gamma}$ secretion when compared with the MVAH4-MVAH4 immunization regimen. The number of $IFN-{\gamma}$-producing splenocytes sensitive to $CD8^+$ T-cell restricted peptides of Ag85B (9-1p and 9-2p) and Th1-related cytokines ($IFN-{\gamma}$ and $TNF-{\alpha}$) in the AdH4-MVAH4 heterologous prime-boost regimen immunization group was significantly higher than that in the other viral vector-based vaccine- and BCG-immunized groups, respectively. These results indicate that an immunization regimen involving AdH4 may have a higher capacity to induce humoral and cellular immune responses against TB in mice than that by regimens containing BCG or MVAH4 alone, and the AdH4-MVAH4 prime-boost regimen may generate an ideal protective effect.

• 한국식품영양과학회지
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• 제44권11호
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• pp.1621-1628
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• 2015

본 연구는 참모자반 조다당 추출물(SFP) 및 에탄올 추출물(SFE)의 면역 활성에 관하여 비교하기 위하여 선천면역계에서 중요한 역할을 수행하는 대표적인 세포인 대식세포와 후천면역계에서 중추적인 역할을 수행하는 비장세포에 각각 SFP와 SFE를 처리하여 각각의 면역세포의 세포 증식률과 사이토카인 분비능에 미치는 영향에 관하여 측정하여 보았다. 대식세포에 SFP 및 SFE를 각각 농도별(12.5, 25, 50, $100{\mu}g/mL$)로 처리하였을 때 두 추출물 모두 대식세포에 대한 세포독성을 유발하지 않았으며, 대식세포의 활성과 밀접한 관련을 가지는 NO, iNOS 및 cytokine의 분비능에 미치는 영향에 관하여 알아본 결과 SFP 처리구에서도 농도 의존적으로 분비능이 증가되는 것으로 관찰되었다. 또한 마우스 비장에서 유리된 비장세포에 SFP 및 SFE를 처리하였을 때 SFP의 처리구에서 비장세포의 증식능 및 면역 활성과 밀접한 관련을 갖는 Th1 type의 cytokine인 IL-2 및 $IFN-{\gamma}$의 분비능이 유의적으로 증가되는 반면, 알레르기를 유도하는 것으로 알려진 Th2 type의 cytokine인 IL-4의 함량에는 영향을 미치지 않는 것으로 관찰되었다. 따라서 참모자반 당류 추출물은 선천면역계와 후천면역계를 담당하는 면역세포인 대식세포 및 비장세포의 활성에 중요한 영향을 미치는 것으로 사료된다.

• IMMUNE NETWORK
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• 제7권3호
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• pp.133-140
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• 2007

Background: It is well established that cross talk between natural killer (NK) cells and myeloid dendritic cells (DC) leads to NK cell activation and DC maturation. In the present study, we investigated whether type 1-polarized DC (DC1) matured in the presence of IFN-${\gamma}$ and type 2-polarized DC (DC2) matured in the presence of PGE2 can differentially activate NK cells. Methods: In order to generate DC, plastic adherent monocytes were cultured in RPMI 1640 containing GM-CSF and IL-4. At day 6, maturation was induced by culturing the cells for 2 days with cytokines or PGE2 in the presence or absence of LPS. Each population of DC was cocultured with NK cells for 24 h. The antigen expression on DC was analyzed by flow cytometry and cytokine production in culture supernatant was measured by ELISA or a bioassay for TNF-${\alpha}$ determination. NK cell-mediated lysis was determined using a standard 4h chromium release assay. Results: DC2, unlike DC1, had weak, if any, ability to induce NK cell activation as measured by IFN-${\gamma}$ production and cytolytic activity. DC2 were weakly stimulated by activated NK cells compared to DC1. In addition, IFN-${\gamma}$-primed mature DC appeared to be most resistant to active NK cell-mediated lysis even at a high NK cell/DC ratio. On the other hand, PGE2-primed DC were less resistant to feedback regulation by NK cells than IFN-${\gamma}$-primed mature DC. Finally, we showed that the differential effect of two types of DC population on NK cell activity is not due to differences in their ability to form conjugates with NK cells. Conclusion: These results suggest that different combinations of inflammatory mediators differentially affect the effector function of DC and, as a result, the function of NK cells, eventually leading to distinct levels of activation in adaptive immunity.

• 혜화의학회지
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• 제16권2호
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• pp.147-169
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• 2007

To evaluate the therapeutic effects of BGG on atopic dermatitis, we investigated the composition of immune cells of lymph node, PBMC and skin of Dermatophagoides farinae-induced NC/Nga mice. The levels of immunoglobulins in serum were analyzed at the protein level and the amount of pathologic cytokines were investigated using CD3/CD28 stimulated splenocytes. The results are summarized below; 1. BGG showed no cytotoxic effect up to $200\;{\mu}g/m{\ell}$ on mLFC in vitro. 2. BGG showed no hepatotoxicity in vivo based on the levels of ALT and AST. 3. Atopic dermatitis was improved through naked eye examination. BGG reduced the skin clinical index from 2.9 to 1.3 (p<0.01). 4. H&E and toluidine blue staining of tissue biopsies revealed that BGG inhibited the infiltration of lymphocytes and mast cells to skin. 5. BGG reduced the number of CD19 positive B cells in PBMCs by 16% (p<0.01), whereas cells were increased by 26% (p<0.05) in lymph nodes. 6. BGG reduced the numbers of B220+/CD23+ cells by 15% (p<0.01) and 33% in PBMCs and lymph node, respectively. 7. BGG reduced the numbers of B220+/IgE+ cells in PBMCs and lymph node by 21% and 33% (p<0.01), respectively. 8. BGG suppressed the levels of IgE (13%, p<0.001) as well as IgM (34%, p<0.001), IgG2a (40%, p<0.001) and IgG2b (26%, p<0.05). 9. BGG reduced the levels of IL-4 and IFN-$\gamma$ by 7% (p<0.05) and 13% (p<0.001) in anti-CD3 and anti-CD28-activated splenocytes, respectively. 10. BGG considerably inhibited the production of TNF-$\alpha$ and IL-6 by 42% (p<0.01) and 15% in the serum, respectively. Based on the results above, we concluded that BGG has therapeutic effects on atopic dermatitis by regulating the differentiation of B cells and isotype switching of IgE. Further investigations on the molecular mechanisms of BGG on atopic dermatitis are anticipated.

• 생명과학회지
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• 제22권10호
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• pp.1307-1315
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• 2012

본 연구에서는 이유자돈 사료에 벤조산 첨가가 위장 내 소화물과 방광 내 뇨의 pH, 장내 유익 미생물 균총수 및 장 조직 내 염증성 싸이토카인 유전자 발현양에 미치는 영향을 조사하였다. 자돈기 5주 동안 이유자돈 사료 내 벤조산 0.3%과 0.5% 첨가는 자돈 위 내 pH는 감소시키지는 않았으나, 방광 내 뇨의 pH 수치는 유의적으로 감소하는 것으로 나타났다. Lactobacillus casei 수는 맹장부위에서 벤조산 0.5% 첨가구와 대조구에서 유의적으로 가장 높게 나타났다. Lactobacillus plantarum 수는 맹장부위에서 벤조산 0.5% 첨가구에서 가장 높게 나타났다. 또한, 직장부위에서 L.plantarum 수는 대조구를 제외하고 모든 처리구에서 유의적으로 높게 나타났다. Bacillus subtillis 수는 맹장부위에서 벤조산0.5% 첨가구와 대조구에서 유의적으로 가장 높게 나타났으며, 직장부위에서는 벤조산0.3%와 0.5% 첨가구에서 가장 높게 나타났다. IL-$1{\beta}$ mRNA 유전자 발현 수준은 소장 상단에서는 벤조산 0.5%와 항생제 처리구에서, 소장중부에서는 벤조산 0.3%와 0.5% 처리구에서, 말단부위에서는 벤조산 0.3% 처리구에서 대조구보다 유의적으로 낮게 나타났다. IL-6 mRNA 유전자 발현 수준은 소장중부에서는 벤조산 0.5% 처리구에서 대조구보다 유의적으로 낮게 나타났다. TNF-${\alpha}$ mRNA 유전자 발현 수준은 소장 중단에서 벤조산 0.5%와 항생제 처리구에서 대조구 보다 낮게 나타났다. 본 실험을 통하여, 자돈 사료 내 benzoic acid 0.5% 첨가는 뇨의 pH 감소와 장내 유익 미생물 균총 수 조절 및 소장 내 면역체계 반응에 긍정적인 영향을 미치므로 자돈 건강을 향상 할 수 있으리라 사료된다.

• KSBB Journal
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• 제30권1호
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• pp.38-43
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• 2015

In this study, we investigated the anti-inflammatory effects of fermented Hizikia fusiformis extracts in lipopolysaccharide (LPS)-stimulated RAW 264.7 mouse macrophages. The fermentation was performed using Lactobacillus casei in mixture of carbon source at $30^{\circ}C$ for 30 days. The sample groups were prepared with/without L. casei group in order to demonstrate the anti-inflammatory activity of fermented H. fusiformis in regard to lactic acid bacteria. As a result, we confirmed the inhibitory effect of H. fusiformis extracts on LPS-stimulated NO production and expression of $TNF{\alpha}$, while it had no regulatory effect on the expression of iNOS, COX-2, $IL-1{\beta}$ and IL-6 as important inflammatory factors. However, L. casei fermented group significantly suppressed the expression of the above factors. In particular, the difference between the two groups in the matter of mRNA expression of iNOS, which is directly associated with NO production, indicated that the fermentation with lactic acid bacteria effectively suppressed NO production by regulating iNOS expression. Also, effective suppression of pro-inflammatory cytokines showed that the fermentation using L. casei may provide an increment towards extraction of active ingredients that are effective anti-inflammatory agents.

• 한국환경보건학회지
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• 제39권1호
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• pp.48-55
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• 2013

Objective: The use of nanoparticle products is expected to present a potential harmful effect on consumers. Also, the lack of information regarding inhaled nanoparticles may pose a serious problem. In this study, we addressed this issue by studying pulmonary toxicity after nasal instillation of Al-NPs in SD rats. Methods: The animals were exposed to Al-NPs at 1 mg/kg body weight (low dose), 20 mg/kg body weight (medium dose) and 40 mg/kg body weight (high dose). To determine pulmonary toxicity, bronchoalveolar lavage (ts.AnBAL) fluid analysis and histopathological examination were conducted in rats. In addition, cell viability was investigated at 24 hours after the treatment with Al-NPs. Results: BAL fluid analysis showed that total cells (TC) count and total protein (TP) concentrations increased significantly in all treatment groups, approximately two to three times. Also, lactate dehydrogenase (LDH) and cytokines such as TNF-alpha and IL-6 dose-dependently increased following nasal instillation of Al-NPs. However, polymorphonuclear leukocytes (PMNs) levels showed no significant changes in a dose dependant manner in BAL fluid. In the cytotoxicity analysis, the treatment of Al-NPs significantly and dose-dependently induced cell viability loss (20 to 30%) and damage of cell membrane (5 to 10%) in rat normal lung epithelial cells (L2). Conclusions: Our results suggest that inhaled Al-NPs in the lungs may be removed quickly by alveolar macrophages with minimal inflammatory reaction, but Al-NPs have the potential to affect lung permeability. Therefore, extensive toxicity evaluations of Al-NPs are required prior to their practical application as consumer products.

• 한국식품영양학회지
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• 제28권2호
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• pp.171-177
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• 2015

본 동물실험은 STZ로 유도한 C57BL/6에게 5% sodium butyrate를 급여했을 때 항당뇨 및 항염증 효과를 연구하고자 하였다. 본 연구에서 STZ로 당뇨를 유발한 마우스에게 5% sodium butyrate를 급여했을 때 체중과 식이섭취량에서는 크게 유의적 차이가 없음을 확인하였다(p<0.05). STZ에 의한 당뇨 쥐는 인슐린의 분비가 감소되면서 당대사의 불균형을 초래하며 간 등이 비대해진다고 알려져 있으나, 본 연구에서는 간의 장기 무게에서는 크게 실험군 간에 유의적인 차이가 없었다(p<0.05). 또한 비장과 흉선의 무게는 0.5% sodium butyrate 첨가 식이군에서 유의적으로 낮아짐을 알 수 있었다(p<0.05). 당뇨병은 염증 상태로서 고혈당으로 인하여 monocyte에서는 여러 염증성 사이토카인이 분비가 활성화된다. TNF-${\alpha}$, IL-6 등은 염증성 사이토카인으로서 혈관염증의 중요한 마커로 인식되고 있고, 당뇨병 환자들은 이러한 염증성 사이토카인이 높은 수준으로 활성화 된다. STZ 처리 시 마우스 혈청에서의 염증성 사이토카인의 분비 및 발현이 증가되었으나, 5% sodium butyrate를 급여했을 때 염증성 사이토카인의 분비 및 발현이 저해됨을 확인할 수 있었다. 본 연구는 sodium butyrate 보충은 당뇨병이 유발된 동물모델에서 혈청지질 농도 및 혈당 조절, 염증 상태를 개선에 다소간의 효과가 있는 것으로 나타났다. 이에 따라 당뇨병과 같은 만성적인 대사질환 개선에 sodium butyrate가 효과적인 식이인자가 될 것으로 생각된다. 그러나 앞으로 더 명확한 효능을 탐색하기 위해서 시료 첨가수준의 다각화 및 여러 가지 보완연구가 필요할 것으로 생각 된다.

• 대한본초학회지
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• 제31권6호
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• pp.1-9
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• 2016

Objectives : Gamisoyosan (GMS) is a useful prescription for treating insomnia, dysmenorrhea and infertility induced by a stress. Also, GMS has been used traditionally to improve systemic circulation and biological energy production. The purpose of this study was to assess the anti-oxidant activity and anti-inflammatory effects of Gamisoyosan Formulation (Soft extract, GMS-SE). Methods : The biological activities such as anti-oxidant and anti-inflammatory effects were measured through cell line-based in vitro assay. We investigated the anti-oxidant properties of GMS-SE on the 1,1-diphenyl-2-picryhydrazyl (DPPH) radical, contents of total flavonoid and polyphenol. GMS-SE compared to butyl hydroxy anizole (BHA). Furthermore, based on this result the anti-inflammatory effects of GMS-SE have verified by mechanism from LPS- treated Raw264.7 macrophages. Results : The anti-oxidant activities of GMS-SE increased markedly, in a dose-dependent manner. The GMS-SE showed significant scavenging activity (GMS-SE $500{\mu}g/m{\ell}$ : $32.77{\pm}1.65%$, GMS-SE $1000{\mu}g/m{\ell}$ : $45.06{\pm}1.04%$ and BHA $100{\mu}g/m{\ell}$ : $39.25{\pm}2.41%$ for DPPH assay). and, The total phenolic compound and flavonoids contents of GMS-SE were $73.93{\pm}6.87{\mu}g/mg$ and $698.75{\pm}6.78{\mu}g/mg$. GMS-SE which is LPS has diminished in the LPS-induced release of inflammatory mediators (NO, iNOS, COX2 and PGE2) and pro-inflammatory cytokines (TNF-${\alpha}$, IL-6 and IL-$1{\beta}$) from the RAW264.7 macrophages. Moreover, GMS-SE inhibited the activation of phosphorylation of p38 and ERK MAPKs by induced LPS. Conclusion : The present results indicate that GMS-SE has an anti-oxidant and anti-inflammatory properties, therefore may be beneficial in diseases which related to oxidative stress-mediated inflammatory disorders.

• 한국약용작물학회지
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• 제17권3호
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• pp.210-216
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• 2009

This study was performed to compare the effects of immuno-modulating activities of Rhodiola sachalinensis A. Bor. fractionized by consecutive solvent separation. The Cytotoxicity of all fractionized extracts on human kidney cell (HEK293) was lower than crude extracts. Generally, the butanol and chloroform extracts showed less cytotoxicity on about 10.07% and 9.67% than the crude extracts. For human immune B and T cell growth, chloroform fraction showed the highest cell growth compared to the control. The secretion of cytokines (IL-6, $TNF-\alpha$) on human B and T cells were increased by adding chloroform extracts. Also, NK cell growth was significantly improved up to nearly 30% by adding the supernatant of B cell medium grown with the chloroform fraction. It was also found that chloroform fraction could yield higher nitric oxide production from macrophage than untreated control cells. Differentiation of HL-60 cells was increased up to 131.9% after treatment with chloroform fraction extracts, compared to the control. These results indicate that the chloroform fraction of R. sachalinensis have high immune activation activity than others fractions and the crude extracts, implying that this chloroform fractions could be used a new functional material.