• The Korean Journal of Physiology and Pharmacology
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• 제16권3호
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• pp.167-174
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• 2012

Natural killer (NK) cells provide one of the initial barriers of cellular host defense against pathogens, in particular intracellular pathogens. Because bone marrow-derived hematopoietic stem cells (HSCs), lymphoid protenitors, can give rise to NK cells, NK ontogeny has been considered to be exclusively lymphoid. Here, we show that porcine c-$kit^+$ bone marrow cells (c-$kit^+$ BM cells) develop into NK cells in vitro in the presence of various cytokines [interleukin (IL)-2, IL-7, IL-15, IL-21, stem cell factor (SCF), and fms-like tyrosine kinase-3 ligand (FLT3L)]. Adding hydrocortisone (HDC) and stromal cells greatly increases the frequency of c-$kit^+$ BM cells that give rise to $CD2^+CD8^+$ NK cells. Also, intracellular levels of perforin, granzyme B, and NKG2D were determined by RT-PCR and western blotting analysis. It was found that of perforin, granzyme B, and NKG2D levels significantly were increased in cytokine-stimulated c-$kit^+$ BM cells than those of controls. And, we compared the ability of the cytotoxicity of $CD2^+CD8^+$ NK cells differentiated by cytokines from c-$kit^+$ BM cells against K562 target cells for 28 days. Cytokines-induced NK cells as effector cells were incubated with K562 cells as target in a ratio of 100 : 1 for 4 h once a week. In results, $CD2^+CD8^+$ NK cells induced by cytokines and stromal cells showed a significantly increased cytotoxicity 21 days later. Whereas, our results indicated that c-$kit^+$ BM cells not pretreated with cytokines have lower levels of cytotoxicity. Taken together, this study suggests that cytokines-induced NK cells from porcine c-$kit^+$ BM cells may be used as adoptive transfer therapy if the known obstacles to xenografting (e.g. immune and non-immune problems) were overcome in the future.

• Journal of Microbiology and Biotechnology
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• 제27권5호
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• pp.1032-1037
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• 2017

The poly-${\gamma}$-$\small{D}$-glutamic acid (PGA) capsule, a major virulence factor of Bacillus anthracis, provides protection of the bacterium from phagocytosis and allows its unimpeded growth in the host. We investigated crosstalk between murine natural killer (NK) cells and macrophages stimulated with the PGA capsule of Bacillus licheniformis, a surrogate of the B. anthracis capsule. PGA induced interferon-gamma production from NK cells cultured with macrophages. This effect was dependent on macrophage-derived IL-12 and cell-cell contact interaction with macrophages through NK cell receptor NKG2D and its ligand RAE-1. The results showed that PGA could enhance NK cell activation by inducing IL-12 production in macrophages and a contact-dependent crosstalk with macrophages.

• Asian Pacific Journal of Cancer Prevention
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• 제14권10호
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• pp.5825-5831
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• 2013

Background/Objectives: Maintenance of cellular function in culture is vital for transfer and development following adoptive immunotherapy. Dual properties of IL-21 in activating T cells and reducing activation induced cell death led us to explore the mechanism of action of IL-21 enhanced proliferation and cytotoxic potential of CIK cells. Method: CIK cells cultured from PBMCs of healthy subjects were stimulated with IL-21 and cellular viability and cytotoxicity to K562 cells were measured. To elucidate the mechanism of action of IL-21, mRNA expression of cytotoxic factors was assessed by RT-PCR and protein expression of significantly important cytotoxic factors and cytokine secretion were determined through flow cytometry and ELISA. Western blotting was performed to check the involvement of the JAK/STAT pathway following stimulation. Results: We found that IL-21 did not enhance in vitro proliferation of CIK cells, but did increase the number of cells expressing the CD3+/CD56+ phenotype. Cytotoxic potential was increased with corresponding increase in perforin ($0.9831{\pm}0.1265$ to $0.7592{\pm}0.1457$), granzyme B ($0.4084{\pm}0.1589$ to $0.7319{\pm}0.1639$) and FasL ($0.4015{\pm}0.2842$ to $0.7381{\pm}0.2568$). Interferon gamma and TNF-alpha were noted to increase ($25.8{\pm}6.1ng/L$ to $56.0{\pm}2.3ng/L$; and $5.64{\pm}0.61{\mu}g/L$ to $15.14{\pm}0.93{\mu}g/L$, respectively) while no significant differences were observed in the expression of granzyme A, TNF-alpha and NKG2D, and NKG2D. We further affirmed that IL-21 signals through the STAT-3 and STAT-5b signaling pathway in the CIK cell pool. Conclusion: IL-21 enhances cytotoxic potential of CIK cells through increasing expression of perforin, granzyme B, IFN-gamma and TNF-alpha. The effect is brought about by the activation of STAT-3 and STAT-5b proteins.

• Journal of Microbiology and Biotechnology
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• 제32권4호
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• pp.397-404
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• 2022

Natural killer (NK) cell activity is more attenuated in hepatocellular carcinoma (HCC) patients than normal. Hypoxic-inducible factor (HIF)-1α is highly expressed in tumors to maintain their metabolism in a hypoxic environment. The expression of HIF-1α in cancers can lead to cell growth, proliferation, invasion/metastasis and immune escape. Although apigenin, a flavonoid, is known to have various biological activities, it has not been demonstrated in NK cell immune activity in HCC cells. In this study, NK-92 cells were directly cocultured with HCC SK-Hep1 cells for 24 h to evaluate NK cell activity in HCC cells or HCC cells expressing HIF-1α by apigenin. NK cell cytotoxicity to HCC cells expressing HIF-1α was significantly increased, and NK cell-activating receptors, NKG2D, NKp30 and NKp44 were highly expressed. The activating effect of apigenin on NK cells substantially induced apoptosis in HCC cells expressing HIF-1α through high expression of CD95L on the surface of NK-92 cells. Moreover, apigenin excellently inhibited the level of TGF-β1 in a coculture of NK cells and HCC cells. In conclusion, apigenin seems to be a good compound that increases NK cell cytotoxicity to HCC cells by controlling HIF-1α expression.

• 대한수의학회지
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• 제60권1호
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• pp.25-32
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• 2020

Natural killer (NK) cells play have a crucial role in the early phase of immune responses against various pathogens. We compared characteristics of canine NK cells against two canine mammary carcinoma cell lines, REM134 and CF41.Mg. REM134 showed higher expression of progesterone receptor, proliferative cell nuclear antigen, Ki67, multiple drug resistance, Bmi-1, c-myc, E-cadherin, and human epidermal growth factor receptor type-2 than that of CF41.Mg. For specific expansion and activation of NK cells, we isolated CD5 negative cells from canine peripheral blood mononuclear cells and co-cultured K562 cells in the presence of interleukin (IL)-2, IL-15, and IL-21 for 21 days. As a result, we found that expression markers of activated NK cells such as NKp30, NKp44, NKp46, NKG2D, CD244, perforin, granzyme B, and tumor necrosis factor alpha were highly upregulated. In addition, we found there was upregulated production of interferon gamma of activated NK cells against target cells such as REM134 and CF41.Mg. Specifically, we observed that cytotoxicity of NK cells against target cells was more sensitively reacted to CF41.Mg than REM134. Based on the results of this study, we recommend the development of an experimental application of CF41Mg, which has not been reported in canine mammary carcinoma research.

• 농업과학연구
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• 제2권1호
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• pp.61-100
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• 1975

수도재배(水稻栽培)에 있어 작중토(作中土)의 유효규산함량(有效珪酸含量), ppm과 유기물함량(有機物含量)% 비(比) $SiO_2$/O.M. 근거(根據)한 N 적량(適量) 추정식(推定式) $Nkg/10a=(4.2+0.096\;SiO_2/O.M.).F$에서 N시비량(施肥量)을 추정(推定) 시용(施用)할 때의 적정(滴定) 가리(加里) 시용량(施用量) 추정식(推定式) $K_2Okg/10a=(K_O/\sqrt{Ca+Mg}-Ks/\sqrt{Ca+Mg})\sqrt{Ca+Mg}{\cdot}47{\cdot}BD$ 단 (但) $K_O/\sqrt{Ca+Mg}=0.03158+0.0007658\;SiO_2/O.M.$ $K_S/\sqrt{Ca+Mg}=Kme/100g/\sqrt{(Ca+Mg)}me/100g$의 타당성(妥當性) 여부(如否)를 판단(判斷)하기 위하여 3수준(水準)의 규회석처리(珪灰石處理)를 주구(主區)로하여 작토중(作土中)의 $SiO_2$/O.M. 비(比)를 증대(增大) 시키고 각주구별(各主區別)로 적정가리시용량(適正加里施用量) 추정식(推定式)에서 추정(推定)한 $K_2O$ 시용량(施用量)과 이에30%를 증비(增肥)하는구(區) 및 적정가리시용추정치(適正加里施用推定値)와는 관계(關係)없이 $K_2O\;8kg/10a$를 시용(施用)하는 3개수준(個水準)의 $K_2O$ 처리구(處理區)를 설정(設定)하여 수도품종(水稻品種) Akibare를 재배(栽培)하는 포장시험(圃場試驗)을 수행(遂行)하고 토양(土壤), 식물체분석(植物體分析) 및 수도(水稻)의 생육(生育)과 수량(收量) 및 수량구성요소(收量構成要素)들을 조사(調査)한 성적(成績)들을 종합검토(綜合檢討)한 결과(結果)를 요약(要約)하면 다음과 같다. 1. 작토중(作土中) 유효규산함량(有效珪酸含量) ppm과 유기물함량(有機物含量)%비(比), $SiO_2$/O.M.에 근거(根據)한 수도(水稻)에 대(對)한 N적량추정식(適量推定式)에서 추정(推定)한 N시용량(施用量)은 수도품종(水稻品種) Akibare의 생육(生育) 및 수량면(收量面)에서 볼 때 과량(過量)이었으며 토양(土壤) 및 식물체(植物體) 분석치(分析値)들에서 검토(檢討)한 결과(結果) $SiO_2$/O.M. 비조절(比調節)의 기본원리(基本原理)는 합당(合當)하나 그 식(式)의 상수(常數) 또는 F값은 변동(變動)되여야 한다고 판단(判斷)되였다. 2. 작토중(作土中)의 K활성도(活性度) 치환성(置換性) 염기 me/100 g비(比) $K/\sqrt{Ca+Mg}$를 조절(調節)하기 위한 $K_2O$ 시비량(施肥量) 추정식(推定式)에서 추정(推定)한 $K_2O$ 시용량(施用量)은 수도품종(水稻品種) Akibare의 생육(生育) 및 수량면(收量面)에서 과량(過量)이였으며 토양(土壤) 및 식물체(植物體) 분석치(分析値)들에서 검토(檢討)한 결과(結果) 조절기준(調節基準)되는 $K_O/\sqrt{Ca+Mg}$의 설정식(設定式) 즉(卽) $K_O/\sqrt{Ca+Mg}=0.03158+0.0007658\;SiO_2/O.M.$의 원리(原理)는 합리적(合理的)이나 기상수(其常數)의 변동(變動)이 필요(必要)하며 $K_S/\sqrt{Ca+Mg}$에 있어서도 토양(土壤)의 K공급능(供給能)을 고려(考慮)할수 있는 새로운 조절기준(調節基準)이 설정(設定)되어야 한다고 판단(判斷)되였다. 3. 수도(水滔)에 대한 $K_2O$ 시용량(施用量) 추정식중(推定式中)의 $K_S/\sqrt{Ca+Mg}$ 조절(調節) 기준(基準)은 K 공급능(供給能)을 대표(代表)할수 있는 작토중(作土中)의 치환성(置換性) Kme/100g에 근거(根據)해서 마음과 같이 설정(設定)함이 보다 합리적(合理的)임을 수도품종(水稻品種) Akibare의 천상수량 $K_2O$ 흡수량면(吸收量面)에서 밝힐수 있었다. 즉 $K_S/\sqrt{Ca+Mg}=0.037+0.78Kme/100g$.