• Nutrition Research and Practice
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• 제13권5호
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• pp.384-392
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• 2019

BACKGROUND/OBJECTIVES: Enteral nutrition formulas with immune-enhancing nutrients, such as n-3 fatty acids, may manage patients' nutritional status and pathophysiological processes. The aim of our study was to investigate natural killer (NK) cell activity alterations and related cytokine changes resulting from feeding with soybean oil-containing enteral nutrition formula (control group) and plant-derived n-3 fatty acid-enriched enteral nutrition formula. SUBJECTS/METHODS: Subjects participated for 14 consecutive days and consumed enteral formula containing canola and flaxseed oil (n3EN, test group) in nonsurgical patients hospitalized for rehabilitation. Blood samples were collected on the first day and 14 days after the consumption of each formula daily, and anthropometric parameters were collected. Hematology and biochemical values were analyzed, and NK cell activities and serum cytokine concentration were measured. A total of sixty subjects were included in the analysis, excluding dropouts. RESULTS: No significant differences were found in biochemical parameters. The n3EN group's NK cell activities at effector:tumor cell ratios of 10:1, 5:1, 2.5:1 and 0.625:1 were significantly higher than those of the control group after two weeks (P < 0.05). However, there were no statistically significant differences in serum cytokine interleukin (IL)-12, $interferon-{\gamma}$, $IL-1{\beta}$, IL-6 and tumor necrosis $factor-{\alpha}$ values between the two groups. CONCLUSIONS: In conclusion, this study elucidates the beneficial effects of plant-derived n-3 fatty acid supplementation in enteral formula on NK cell activity.

• 대한임상검사과학회지
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• 제50권3호
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• pp.261-266
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• 2018

만성C형 간염환자 160예를 대상으로 immunoblot방법과 RT-PCR-hybridization법을 실시하여 임상검사의 유용성을 알아보았다. 양성 150예 중 133예만 RT-PCR-hybridization 법에서 양성을 나타내어 immunoblot법의 진양성율은 88.6%를 나타났으며 두 방법 간의 일치율은 89.3%를 나타났다. 한국인의 HCV 아형을 알기 위하여 혈청형과 유전형을 실시하였다. HCV혈청형 1형과 2형 그리고 1b와 2a유전형이 가장 많은 C형간염 감염원으로 나타났다. 49예를 혈청형과 유전형을 서로 비교한 결과, 혈청형 검사에서 28예(57.1%)가 1형, 21예(42.9%)가 2형으로 나타났으며 유전자형 검사에서 1b형이 25예(51.0%), 1b/2b를 나타낸 예가 1예(2.0%), 2a형이 17예(34.7%), 2a/2c를 나타낸 예가 4예(8.2%), 그리고 2b형이 2예(4.1%)로 나타났다. 본 연구에서는 HCV 간이검출법으로는 immunoblot방법이 유용하며, immunoblot방법 양성 확진검사로는 RT-PCR-hybridization법을 실시하였다. 혈청형이 C형간염의 치료 나 진행과정을 관찰하기 위해서는 유전형보다 유용하나 인터페론 치료효과는 1형과 2형 혈청형별에 따라 차이를 보이지 않았다.

• Journal of Ginseng Research
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• 제39권1호
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• pp.29-37
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• 2015

Background: Panax ginseng (i.e., ginseng) root is extensively used in traditional oriental medicine. It is a modern pharmaceutical reagent for preventing various human diseases such as cancer. Ginsenosidesd-the major active components of ginsengd-exhibit immunomodulatory effects. However, the mechanism and function underlying such effects are not fully elucidated, especially in human monocytes and dendritic cells (DCs). Methods: We investigated the immunomodulatory effect of ginsenosides from Panax ginseng root on $CD14^+$ monocytes purified from human adult peripheral blood mononuclear cells (PBMCs) and on their differentiation into DCs that affect $CD4^+$ T cell activity. Results: After treatment with ginsenoside fractions, monocyte levels of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, and IL-10 increased through phosphorylation of extracellular signal-regulated kinase (ERK)1/2 and c-Jun N-terminal kinase (JNK), but not p38 mitogen-activated protein kinase (MAPK). After treatment with ginsenoside fractions, TNF-${\alpha}$ production and phosphorylation of ERK1/2 and JNK decreased in lipopolysaccharide (LPS)-sensitized monocytes.We confirmed that DCs derived from $CD14^+$ monocytes in the presence of ginsenoside fractions (Gin-DCs) contained decreased levels of the costimulatory molecules CD80 and CD86. The expression of these costimulatory molecules decreased in LPS-treated DCs exposed to ginsenoside fractions, compared to their expression in LPS-treated DCs in the absence of ginsenoside fractions. Furthermore, LPS-treated Gin-DCs could not induce proliferation and interferon gamma (IFN-${\gamma}$) production by $CD4^+$ T cells with the coculture of Gin-DCs with $CD4^+$ T cells. Conclusion: These results suggest that ginsenoside fractions from the ginseng root suppress cytokine production and maturation of LPS-treated DCs and downregulate $CD4^+$ T cells.

• 한국미생물·생명공학회지
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• 제49권1호
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• pp.75-87
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• 2021

Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp₄₅ signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SP_Usp45-IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC₅₀ values of 33.22 ㎍/ml and 127.2 ㎍/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 ㎍/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 ㎍/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codon-optimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

• 한국식품저장유통학회지
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• 제31권2호
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• pp.307-314
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• 2024

닥나무는 항산화, 항암 효과뿐 아니라 천연 미백기능성분으로 인정받아 화장품 원료로 사용되며 닥나무의 잎과 가지는 식약처에 등재된 식용이 가능한 원료이다. 닥나무는 아시아에서 종이 제조 및 한의학적 용도로 사용되었고 항당뇨 등의 효능이 있으며, 다양한 플라보노이드와 알칼로이드를 포함하는 것으로 알려져 있다. 본 연구에서는 닥나무 추출물의 피부 염증성 알러지 반응에 대한 효능을 평가하기 위하여, HaCaT 각질형성세포의 피부염증 억제와 RBL-2H3 세포의 알러지 반응 억제와 관련된 인자들에 미치는 영향에 관하여 연구하였다. HaCaT 및 RBL-2H3 세포에 대한 70% 에탄올 추출물의 세포독성은 나타나지 않았다. HaCaT 세포에서 TNF-𝛼와 IFN-𝛾의 자극으로 케모카인(TARC, MDC, RANTES) 생성이 증가하였으며, 시료처리에 따라 농도의존적으로 유의성 있게 감소하였다. 한편, IgE 처리로 활성화된 RBL-2H3 세포에서 증가하는 𝛽-hexosaminidase 방출과 염증성 사이토카인 TNF-𝛼, IL-4 생성이 시료 처리로 유의적으로 감소하는 것을 확인하였다. 따라서 닥나무 가지 추출물은 알러지 염증반응 완화 효과를 갖는 천연물 화장품 및 식품 원료로 활용이 가능할 것으로 예상된다.

• Tuberculosis and Respiratory Diseases
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• 제41권2호
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• pp.87-96
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• 1994

연구배경 : 종양괴사인자(tumor necrosis factor ; TNF)는 여러 암세포에 대해서 세포독성을 보임이 알려져 있다. 그리고, 최근 분자생물학적 발전에 힘입어 TNF 유전자를 암세포에 이입하고 발현시키는 연구가 그 동안 진행되었다. 이들 연구의 목적은, TNF를 전신적으로 쓸 경우 전신 부작용이 심하여 인체에 쓸 수 없는 현 단계에서 암세포 자체에서 TNF를 만들어 내어서 암세포 주위에서만 작용하게 함으로써 연체에 미치는 전신독성을 최소한으로 줄이고 암세포를 사멸시키는 것이었다. 암세포에 TNF 유전자를 이입함으로써 예상되는 항암효과가 성공을 거두기 위해서는 첫째, TNF 유전자가 이입된 암세포에서 분비된 TNF가 주위 암세포를 성공적으로 사멸시켜야 하고 둘째는 분비된 TNF가, TNF 유전자가 이입된 암세포 자신을 사멸시켜야 한다. 본 연구는 이 중 두번째 기전, 즉 TNF 유전자가 이입된 암세포가 자신이 분비한 TNF에 의해서 사멸되거나 또는 세포 독성이 나타날 수 있는가를 검증하는데 목적을 두었다. 방법 : TNF에 감수성을 보이는 인체의 중피종 세포주인 NCI-H2058과 생쥐 섬유육종 세포주인 WEHI164에 TNF-$\alpha$ 유전자를 retroviral vector를 이용하여 이입하고 TNF를 발현을 시도하였다. DNA 수준과 단백질 수준에서 TNF-$\alpha$ 유전자가 제대로 이입되어 발현되는지 PCR과 ELISA 및 bioassay(MTT assay)로 확인하였다. 그리고, TNF 유전자가 이입된 세포주가 자신이 분비하는 TNF에 사멸되는지 아니면 생존하는지 MTT(dimethylthiazolyl diphenyltetrazolium) assay로 알아보았다. 그리고, 만일 TNF 유전자가 이입된 암세포가 자신이 분비한 TNF에 사멸되지 않고 생존할 경우, TNF 유전자가 이입된 NCI-H2058-TNF와 WEHI164-TNF 암세포주가 외부에서 준 TNF에도 내성을 보이는지도 추가로 MTT assay 방법으로 확인해 보았다. 결과 : 1) TNF-$\alpha$ 유전자 이입 및 발현 확인 PCR을 시행한 결과, TNF 유전자가 이입된 NCI-H2058-TNF와 WEHI164-TNF 세포주는 790 base pair 크기의 진한 DNA band를 보인 반면 각각의 모세포주에서는 보이지 않아서 retroviral vector를 이용한 유전자 이입이 DNA 수준에서 이루어졌음을 확인할 수 있었다. 그리고, NCI-H2058-TNF와 WEHI164-TNF의 상층 배양액의 TNF양을 ELISA와 bioassay(MTT assay)로 측정한 결과, 생물학적 활성을 지닌 TNF를 각각 $23.6{\pm}0.84ng$/24h/$10^6cells$, $12.2{\pm}0.36ng$/24h/$10^6cells$ 생산함을 알 수 있었다. 2) TNF 유전자 이입 전후, 암세포의 TNF에 대한 감수성 비교 TNF 유전자 이입 전후의 TNF에 대한 세포주의 감수성(세포사망율)을 TNF의 농도 변화에 따라 비교한 결과, NCI-H2058의 경우 TNF 농도 100ng/ml에서 모세포는 $25{\pm}3%$의 세포독성을 보인 반면 TNF 유전자 이입 후에는 $3{\pm}2%$의 세포독성을 보여 통계적으로 유의한 차이가 있었다(p<0.01). 그리고, WEHI-164의 경우도 TNF 농도 100ng/ml에서 모세포는 $73{\pm}5%$의 세포독성을 보인 반면 TNF 유전자 이입 후에는 $3{\pm}2%$의 세포독성을 보여 통계적으로 유의한 차이가 있었다(p<0.01). 결론 : TNF에 감수성을 보이는 암세포주인 NCI-H2058과 WEHI164에 TNF 유전자 이입을 시행하고 TNF가 발현되게 하였을 때, TNF 유전자를 이입받은 두 암세포주 모두에서 자신이 생산해 내는 TNF에 내성을 보여 생존하였다. 뿐만 아니라 생존한 이들 세포는 외부에서 준 TNF에 대해서도 내성을 보였다. 따라서, 암세포에 대한 TNF 유전자 이입을 통한 유전자 요법이 성공을 거두려면 유전자 이입된 세포에서 분비하는 TNF의 면역 세포 동원 방법 등의 간접적인 항암기전이 필요할 것으로 생각된다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권12호
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• pp.1753-1759
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• 2008

Forty weanling piglets ($5.6{\pm}0.5kg$ and 26 to 30 d of age) were used in a 28-d experiment to determine the effects of ${\beta}$-glucan from Paenibacillus polymyxa and L-theanine on growth performance. Piglets were randomly allotted to four groups (n = 10, 2 animals per pen) provided with the basal feed (control), ${\beta}$-glucan 400 mg/kg feed, L-theanine 80 mg/kg feed or ${\beta}$-glucan plus l-theanine (combination of the above-mentioned concentrations). Body weight and feed consumption were recorded during four weeks. Subsequently, the immunomodulatory effects of ${\beta}$-glucan and L-theanine were investigated for lipopolysaccharide (LPS)-induced cytokine production in vitro and in vivo on day 28. Although there were no significant differences in the growth performances among the treatment groups, ${\beta}$-glucan plus L-theanine had 5.6% greater ADG (p = 0.074) on day 21 to 28. ${\beta}$-Glucan alone or plus L-theanine increased interleukin (IL)-10 levels and decreased interferon (IFN)-$\gamma$ and tumor necrosis factor (TNF)-${\alpha}$ levels in cultured medium by LPS treatment (p<0.05). Plasma IL-10 levels were also increased in the piglets fed with ${\beta}$-glucan alone or plus L-theanine after LPS challenge ($25{\mu}g/kg$, i.p.), whereas plasma IFN-$\gamma$ and TNF-${\alpha}$ levels were decreased (p<0.05). The levels of IFN$\gamma$ in piglets fed with ${\beta}$-glucan plus L-theanine showed the greatest inhibition after LPS challenges. In conclusion, treatment of ${\beta}$-glucan alone or plus L-theanine might lessen inflammatory responses against Gram-negative bacterial infection via the inhibition of pro-inflammatory cytokine production and enhancement of anti-inflammatory cytokine production. Further studies are needed to determine an optimal concentration of ${\beta}$-glucan and L-theanine for improved growth performance.

• 대한한의학방제학회지
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• 제20권2호
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• pp.13-28
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• 2012

Objectives : The present study aimed to investigate the pharmacological activity of water and ethanol (EtOH) extracts from Socheongnyong-tang (SCNT) on inflammation and its related disease. Methods : The cells were treated with nontoxic concentrations of water and EtOH extract from SCNT in BEAS-2B, HaCaT, RAW 264.7 and 3T3-L1 cells. These cells were stimulated by tumor necrosis facter (TNF)-${\alpha}$, TNF-${\alpha}$/interferon (IFN)-${\gamma}$, and lipopolysaccharide (LPS), respectively. 3T3-L1 cells were differentiated by insulin. After incubation, supernatant were collected and biological indicator measured by enzyme-linked immunosorbent assay. Results : Our results indicate that the water and EtOH extract of SCNT significantly inhibited the production of regulated on activation normal T-cell expression and secreted (RANTES) by treatment of TNF-${\alpha}$ in BEAS-2B cell, and significantly reduced the production of RANTES and macrophage-derived chemokine increased by treatment of TNF-${\alpha}$/IFN-${\gamma}$ in HaCaT cell. Moreover, those extracts significantly decreased the activity of nitric oxide and prostaglandin $E_2$ in LPS-induced RAW 264.7, and significantly inhibited the increased activity of glycerol-3-phosphate dehydrogenase and expression of leptin induced by differentiation in 3T3-L1 cell. Conclusions : These results indicate that both water and EtOH extract of SCNT has powerful effects on inflammation and its related disease. Therefore, SCNT can be developed as a potential pharmacological agent related various diseases. Although the significant effects were observed in both SCNT water and EtOH extract, the EtOH extract was more effective on most experiments than its water extract. Taken together, these findings indicate that the SCNT EtOH extract may have more potential pharmacological agent.

• Journal of Microbiology and Biotechnology
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• 제27권6호
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• pp.1071-1077
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• 2017

A rise in the occurrence of allergic diseases is attributed to the dysregulated balance of type 1/type 2 immunity, where type 2 T-helper (Th2) cells predominate over type 1 T-helper (Th1) cells, leading to an abnormally increased production of IgE in response to unharmful antigens. Kimchi, a traditional Korean fermented food, is a rich source of beneficial lactic acid bacteria. In this study, we investigated the ability of Enterococcus faecium FC-K derived from kimchi to induce type I immunity in the presence of Th2 polarizing conditions in vitro and in vivo. Stimulation of mouse peritoneal macrophages with E. faecium FC-K induced the production of tumor necrosis factor alpha, interleukin (IL)-6, and IL-12. Under the in vitro Th2 conditions in which splenic T cells were activated in the presence of IL-4, E. faecium FC-K enhanced the ability of T cells to produce interferon $(IFN)-{\gamma}$. Using the ovalbumin (OVA)-induced allergy model, male BALB/c mice receiving E. faecium FC-K reduced the serum level of total IgE, but not that of OVA-specific IgE. Furthermore, the population of activated splenic B cells during OVA immunization was decreased in E. faecium FC-K-treated mice, accounting for a reduction of total IgE in the serum. Restimulating splenocytes from OVA-immunized mice with OVA ex vivo resulted in an increased production of $IFN-{\gamma}$, with no effect on IL-4, in E. faecium FC-K-treated mice. These observations provide the evidence that E. faecium FC-K can be a beneficial probiotic strain that can modulate the Th2-mediated pathologic response.

• Journal of Korean Medical Science
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• 제33권52호
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• pp.336.1-336.12
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• 2018

Background: We aimed to investigate mucosal immunity related to forkhead box P3 ($FOXP3^+$) regulatory T (Treg) cells, T helper 17 (Th17) cells and cytokines in pediatric inflammatory bowel disease (IBD). Methods: Mucosal tissues from terminal ileum and colon and serum samples were collected from twelve children with IBD and seven control children. Immunohistochemical staining was done using anti-human FOXP3 and anti-$ROR{\gamma}t$ antibodies. Serum levels of cytokines were analyzed using a multiplex assay covering interleukin $(IL)-1{\beta}$, IL-4, IL-6, IL-10, IL-17A/F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, interferon $(IFN)-{\gamma}$, soluble CD40L, and tumor necrosis factor-${\alpha}$. Results: $FOXP3^+$ Treg cells in the lamina propria (LP) of terminal ileum of patients with Crohn's disease were significantly (P < 0.05) higher than those in the healthy controls. $ROR{\gamma}t^+$ T cells of terminal ileum tended to be higher in Crohn's disease than those in the control. In the multiplex assay, serum concentrations (pg/mL) of IL-4 ($9.6{\pm}1.5$ vs. $12.7{\pm}3.0$), IL-21 ($14.9{\pm}1.5$ vs. $26.4{\pm}9.1$), IL-33 ($14.3{\pm}0.9$ vs. $19.1{\pm}5.3$), and $IFN-{\gamma}$ ($15.2{\pm}5.9$ vs. $50.2{\pm}42.4$) were significantly lower in Crohn's disease than those in the control group. However, serum concentration of IL-6 ($119.1{\pm}79.6$ vs. $52.9{\pm}39.1$) was higher in Crohn's disease than that in the control. Serum concentrations of IL-17A ($64.2{\pm}17.2$ vs. $28.3{\pm}10.0$) and IL-22 ($37.5{\pm}8.8$ vs. $27.2{\pm}3.7$) were significantly higher in ulcerative colitis than those in Crohn's disease. Conclusion: Mucosal immunity analysis showed increased $FOXP3^+$ T reg cells in the LP with Crohn's disease while Th17 cell polarizing and signature cytokines were decreased in the serum samples of Crohn's disease but increased in ulcerative colitis.