• Asian Pacific Journal of Cancer Prevention
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• 제15권18호
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• pp.7995-8001
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• 2014

Interferon-gamma (IFN-${\gamma}$) has been used to treat various malignant tumors. However, the molecular mechanisms underlying the direct anti-proliferative activity of IFN-${\gamma}$ are poorly understood. In the present study, we examined the in vitro antitumor activity of IFN-${\gamma}$ on two human non-small-cell lung carcinoma (NSCLC) cell lines, H322M and H226. Our findings indicated that IFN-${\gamma}$ treatment caused a time-dependent reduction in cell viability and induced apoptosis through a FADD-mediated caspase-8/tBid/mitochondria-dependent pathway in both cell lines. Notably, we also postulated that IFN-${\gamma}$ increased indoleamine 2,3-dioxygenase (IDO) expression and enzymatic activity in H322M and H226 cells. In addition, inhibition of IDO activity by the IDO inhibitor 1-MT or tryptophan significantly reduced IFN-${\gamma}$-induced apoptosis and death receptor 5 (DR5) expression, which suggests that IDO enzymatic activity plays an important role in the anti-NSCLC cancer effect of IFN-${\gamma}$. These results provide new mechanistic insights into interferon-${\gamma}$ antitumor activity and further support IFN-${\gamma}$ as a potential therapeutic adjuvant for the treatment of NCSLC.

• Biomolecules & Therapeutics
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• 제20권3호
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• pp.293-298
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• 2012

The purpose of this study was to investigate the modification of expression and functionality of the drug transporter P-glycoprotein (P-gp) by tumor necrosis factor-alpha (TNF-${\alpha}$) and interferon-gamma (IFN-${\gamma}$) at the blood-brain barrier (BBB). We used immortalized human brain microvessel endothelial cells (iHBMEC) and primary human brain microvessel endothelial cells (pHBMEC) as in vitro BBB model. To investigate the change of p-gp expression, we carried out real time PCR analysis and Western blotting. To test the change of p-gp activity, we performed rhodamin123 (Rh123) accumulation study in the cells. In results of real time PCR analysis, the P-gp mRNA expression was increased by TNF-${\alpha}$ or IFN-${\gamma}$ treatment for 24 hr in both cell types. However, 48 hr treatment of TNF-${\alpha}$ or IFN-${\gamma}$ did not affect P-gp mRNA expression. In addition, co-treatment of TNF-${\alpha}$ and IFN-${\gamma}$ markedly increased the P-gp mRNA expression in both cells. TNF-${\alpha}$ or IFN-${\gamma}$ did not influence P-gp protein expression whatever the concentration of cytokines or duration of treatment in both cells. However, P-gp expression was increased after treatments of both cytokines together in iHBMEC cells only compared with untreated control. Furthermore, in both cell lines, TNF-${\alpha}$ or IFN-${\gamma}$ induced significant decrease of P-gp activity for 24 hr treatment. And, both cytokines combination treatment also decreased significantly P-gp activity. These results suggest that P-gp expression and function at the BBB is modulated by TNF-${\alpha}$ or/and IFN-${\gamma}$. Therefore, the distribution of P-gp depending drugs in the central nervous system can be modulated by neurological inflammatory diseases.

• 대한의생명과학회지
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• 제5권1호
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• pp.85-93
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• 1999

마우스 간 세포주인 BNL CL.2의 시험관내 배양에서 혈청과 IFN-$\gamma$가 세포주의 nitric oxide (NO)생성과 세포 손상에 미치는 영향을 알아보기 위한 실험을 하였다. 혈청이 공급된 배양에서 IFN-$\gamma$에 의한 세포 생존율은 거의 변동이 없었으나, 혈청을 제거한 배양에서는 약 65%의 생존율이 유지되었으며, NO생성 억제제인 N$^{G}$-monomethy-L-arginine (NMA)의 첨가는 농도 의존적으로 세포의 생존율을 감소시켰다. 혈청이 제거된 BNL CL.2 세포주는 IFN-$\gamma$ 단독 처리에서도 NO를 생성할수 있었으며, IFN-$\gamma$와 lipopolysaccharide (LPS)의 복합 처리는 세포주의 NO 생성을 상승적으로 증가시 켰다. 또한 protein tyrosine kinase (PTK) inhibitor인 herbimycin A와 genistein에 의해서 NO 생성이 억제되어 PTK의 활성이 혈청이 고갈된 BNL CL.2세포에서 NO의 생성에 중요한 역할을 담당하고 있기 때문으로 판단된다. IFN-$\gamma$의 독성은 혈청을 제거시킬 때 NO 생성 억제제에 상승적으로 간세포를 손상시키며, 이때 NO가 IFN-$\gamma$에 의해 유도된 손상을 어느 정도 억제시키는 것을 알 수 있었다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권14호
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• pp.5635-5641
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• 2015

Background: Cirrhosis is regarded as a possible end stage of many liver diseases, including viral infection. It occurs when healthy liver tissue becomes damaged and is replaced by scar tissue and finally may lead to hepatocellular carcinoma. Interferons (IFNs)are two general categories, type I and II. Type I includes one beta interferon and over 20 different alpha interferons. Alpha interferons are very similar in how they work, interacting with other proteins on cells like receptors. The main objective of this study was to compare Mx gene productivity post different cell line treatment with imported and Egyptian biosimilar locally produced IFNs, as well as the efficacy of those tested IFNs. Also, an assessment was made of sensitivity of different cell lines as alternatives to that recommended for evaluation of antiviral activity. Materials and Methods: Different cell lines (Vero, MDBK and Wish) were employed to evaluate cytotoxicity using the MTT assay. Antiviral activity was evaluated compared with standard IFN against VSV, Indiana strain -156, on tested rh-IFNs (imported; innovated and Egyptian biosimilar locally produced IFNs) in the pre-treated cell lines previously mentioned. The virus was propagated in the Wish cell line as recommended. Finally we estimated up-regulation of the Mx gene as a biomarker. Results: Data recorded revealed that test IFNs were safe in test cell lines. Viability was around 100%. Locally tested interferon did not realize the international potency limits, while the imported one was accepted compared with the standard IFN. These results were the same either using infectivity titer reduction assay or crystal violet staining of residual non- infected cells. Mx protein production was cell type related and confirmed by the detected Mx gene expressed in imported and locally produced IFN pre-treated cell lines. The expression of the gene was arranged in the order of Vero> wish > MDBK for the imported IFN, while for the Egyptian biosimillar locally produced one it was MDBK> Vero> wish. With regard to the antiviral activity there was a significant difference of imported IFN potency compared with the locally produced IFN (P<0.05), the IFN potential (antiviral activity) was not cell line related and showed non-significant difference for each separate product. Conclusions: Vero cells can be used as an alternative cell line for evaluation of IFN potency in case of unavailable USP recommended cell lines. Alternative potency evaluation assay could be used and proved significant difference in IFN potency in case of local and imported agents. Evaluation of antiviral activity could be used in parallel to viral infectivity reduction assay for better accuracy. Mx gene can be used as a marker for IFN potential.

• 농업과학연구
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• 제12권1호
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• pp.153-161
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• 1985

고단위(高單位)의 porcine leucocyte interferon(For IFN-${\alpha}$)을 대량생산(大量生産)하기 위한 기초연구(基礎硏究)로써 백혈구(白血球) 분리법(分離法), 백혈구(白血球)의 성상(性狀) 및 inducer에 따른 IFN 생산효과(生産效果) 시험(試驗)을 하였고 Por IFN-${\alpha}$의 역가측정(力價測定)을 micro-system에 확립(確立)하기 위해 시험(試驗)을 수행(遂行)하였던 바 다음과 같은 결과(結果)를 얻었다. 1. 돼지 말초백혈구(末梢稍白血球)를 분리(分離)하기 위해서 4 가지 방법(方法)을 응용(應用)하던 바 백혈구(白血球) 분리법(分離法)은 buffy coat 方法에서 가장 높았으나 적혈구(赤血球) 혼입율(混入率)이 높았고 Ficoll-paque 방법(方法)에서는 백혈구(白血球) 분리율(分離率)은 중등도(中等度)였으나 적혈구(赤血球) 혼입율(混入率)은 가장 낮았다. 2. 말초백혈구(末梢白血球)와 비장세포(脾臟細胞)의 활성도(活性度)는 IFN 생산(生産)에 영향(影響)을 주었으며 세포활성도(細胞活性度)가 높을수록 IFN 생산효능(生産效能)이 높았으며, 적혈구(赤血球)의 혼입도(混入度)가 높을수록 IFN 생산효능(生産效能)은 낮았다. 말초백혈구(末梢白血球)는 비장세포(脾臟細胞)에 비해 1.7배(倍)의 높은 IFN 생산효과(生産效果)가 인정(認定)되었다. 3. Inducer로 사용(使用)된 Sendai virus(Cantell 주(株))는 Newcastle disease virus(Lasota 주(株)) 보다 IFN 생산효과(生産效果)가 5.5 배(倍) 높았다. 4. 본(本) 시험(試驗)에서 확립(確立)된 micro-IFN assay 법(法)의 재현성(再現性)을 검사(檢査)한 바 4회반복시험(回反復試驗)에서 편차(偏差)는 0.05~0.21로 나타났다. 5. 시험생산(試驗生産) 9롯트의 IFN 역가(力價)는 330 내지 4,700 unit/ml로써 평균(平均) 1,200 unit/ml이었다.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 2006년도 춘계학술대회
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• pp.83-88
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• 2006

In this study we examined the potential for the synergistic augmentation of the activity of inflammatory mouse peritoneal macrophages by stimulation with RGAP combined with IFN-$\gamma$. The moderate augmentative effect induced by preincubation with RGAP was observed in the production of IL-1, IL-6 and NO but not TNF-$\alpha$. In addition, IFN-$\gamma$ had a low activating effect. Following preincubation with both RGAP A and IFN-$\gamma$, a marked enhancement of secretory activity and tumoricidal activity was noted in mouse peritoneal macrophages. Treatment of peritoneal macrophage with combination increased the generation of IL-1, IL-6, TNF-$\alpha$ and NO, whereas the production of reactive oxygen species were not altered. These results demonstrate the synergistic effects on macrophage function of RGAP in combination with IFN-$\gamma$ and suggest that the ability of IFN-$\gamma$ to prime macrophages to produce secretory molecules in response to RGAP may have implications for immunotherapy with this combination.

• Parasites, Hosts and Diseases
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• 제38권2호
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• pp.85-90
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• 2000

To assess the relationship between the changes of cellular components and the production of Th 1 cytokine in the immune tissue, inbred C57BL/6 mice were orally infected with 40 cysts of 76K strain of Toxoplosma gondii. The sequential change of cell differentials and $IFN-{\gamma}$ production of splenocytes were analyzed by Diff-Quik stain and RT-PCR. There were no significant proportional changes of cellular components of splenocytes until day 4 postinfection (Pl) as compared to those of day 0, and the relative percentage of macrophages and neutrophils/eosinophils increased significantly (p<0.01) thereafter. The expression of $IFN-{\gamma}$ mRNA of $CD3^{-}$ cells was observed from day 1 Pl at a low level. However, $IFN-{\gamma}$ production of $CD3^{+}$ cells increased significantly from day 4 Pl (p<0.01) which progressively increased thereafter. These findings provide the relative percentages of granulocytes and macrophages were increased in conjunction with increase of total number of splenocytes after oral infection with T. gondii in the susceptible murine hosts, and lymphocytes were the major cellular components and the important source of $IFN-{\gamma}$.

• IMMUNE NETWORK
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• 제12권4호
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• pp.148-154
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• 2012

Previously, we have reported that high mobility group box 1 (HMGB1), a proinflammatory mediator in sepsis, is released via the IFN-${\beta}$-mediated JAK/STAT pathway. However, detailed mechanisms are still unclear. In this study, we dissected upstream signaling pathways of HMGB1 release using various molecular biology methods. Here, we found that calcium/calmodulin-dependent protein kinase (CaM kinase, CaMK) is involved in HMGB1 release by regulating IFN-${\beta}$ production. CaMK inhibitor, STO609, treatment inhibits LPS-induced IFN-${\beta}$ production, which is correlated with the phosphorylation of interferon regulatory factor 3 (IRF3). Additionally, we show that CaMK-I plays a major role in IFN-${\beta}$ production although other CaMK members also seem to contribute to this event. Furthermore, the CaMK inhibitor treatment reduced IFN-${\beta}$ production in a murine endotoxemia. Our results suggest CaMKs contribute to HMGB1 release by enhancing IFN-${\beta}$ production in sepsis.

• Animal cells and systems
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• 제8권2호
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• pp.117-123
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• 2004

The purpose of this study was to analyze inhibitory effects of anti-4-1BB monoclonal antibody on melanoma metastasis The 4-1BB (CD137) T cell molecule is a member of the TNF receptor family and its activation by either 4-1BB ligand or antibody induces T cell activation and growth. In the present study, administration of anti-4-1BB mAb induced inhibition of melanoma metastasis. Agonistic anti-4-1BB mAb induced not only CD$8^+$4-1BBT cells but also CD$8^+$IFN-${\gamma}$$^{+}$ T cell population. In the presence of anti-CD3 antibody, lymphocytes produced high levels of IFN-${\gamma}$ and low levels of IL-4 in anti-4-1BB mAb treated group. Exposure of melanoma cells to IFN-${\gamma}$ induced expression of MHC-I molecules. Thus, the increase in number of CD$8^+$T cells and enhanced MHC-I expression on B16F10 cells by augmented IFN-${\gamma}$ production in response to anti-4-1BB mAb may result in suppression of tumor growth and metastasis.s.

• 한국식품영양학회지
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• 제20권3호
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• pp.259-264
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• 2007

Ginger(Zingiber officinale Roscoe) has been used as a raw material in many various traditional preparations since the ancient times. As a component of traditional health products, ginger is known to be effective as an appetite enhancer, and has anti-cold and anti-inflammatory activities. This study was performed to investigate the immunomodulative effects of Zingiber officinale Roscoe in mice, using ex vivo experiments. In order to elucidate ginger's immunomodulative effects of Ginger, water extracts were orally administered to mice, and isolated macrophages were used as the experimental model. To identify the ex vivo effects, six to seven week old Balb/c mice were fed a chow diet ad libitum and the ginger water extracts were administered orally every other day for two or four weeks at two different concentrations(50 and 500 mg/kg b.w.). The results show that IL-IO and $IFN-{\gamma}$ were detected in the 500 mg/kg b.w. supplemented group with LPS stimulation in all cases. Also, the $IFN-{\gamma}$ /IL-10 ratio ranged from 3~5 with mitogen stimulation such as Con A and LPS. In conclusion, this study suggests that ginger extracts may enhance the immune function by regulating the cytokine(IL-10 and $IFN-{\gamma}$) production capacity of activated macrophages in mice.