• 제목/요약/키워드: cell enzyme-linked immunosorbent assay

검색결과 297건 처리시간 0.029초

Fusarium sp.가 생성하는 zearalenone에 대한 단크론성 항체생산 (Production of Monoclonal Antibody against Zearalenone Produced by Fusarium sp.)

  • 강성조;정덕화;강진순
    • 한국식품과학회지
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    • 제30권6호
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    • pp.1409-1414
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    • 1998
  • Zearalenone에 특이한 단크론성 항체를 개발하기위하여 형질세포종세포인 myeloma cell $(P3{\times}63Ag.V653)$과 zearalenone-oxime-bovine serum albumin (BSA) 결합체를 항원으로 면역시킨 BALB/c female mice의 비장세포를 융합시켜 hybridoma cell을 생산하였다. 그들의 항체가를 indirect competitive ELISA법으로 측정한 결과 zearalenone에 대하여 높은 친화력을 갖는 5세포주를 얻었다. 그중 Z-2-M26 hybridoma cell line이 생산하는 단크론성항체는 특히 zearalenone과 민감하게 반응하였고, ${\alpha}-zearalenol$과도 약간의 교차반응을 보였으나 ${\beta}-zearalenol,\;{\alpha}-zearalenol,\;{\beta}-zearalenol$ 및 DON과는 거의 반응하지 않았다. 따라서 본 실험에서 개발된 항체는 앞으로 zearalenone에 대한 민감하고 정량적인 효소면역측정법의 개발을 위한 중요한 면역시약으로 사용될 것으로 생각한다.

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Preliminary Study on Natural Killer Cell Activity for Interfer-on-Gamma Production after Gamma Knife Radiosurgery for Brain Tumors

  • Park, Kawngwoo;Jeong, Sang Soon;Kim, Jung Hoon;Chung, Hyun-Tai;Lee, Eun Jung;Moon, Hyo Eun;Park, Kwang Hyon;Kim, Jin Wook;Park, Hye Ran;Lee, Jae Meen;Lee, Hye Ja;Kim, Hye Rim;Cho, Yong Hwan;Paek, Sun Ha
    • Journal of Korean Neurosurgical Society
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    • 제65권6호
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    • pp.861-867
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    • 2022
  • Objective : High-dose radiation is well known to induce and modulate the immune system. This study was performed to evaluate the correlation between clinical outcomes and changes in natural killer cell activity (NKA) after Gamma Knife Radiosurgery (GKS) in patients with brain cancer. Methods : We performed an open-label, prospective, cross-sectional study of 38 patients who were treated with GKS for brain tumors, including metastatic and benign brain tumors. All of the patients underwent GKS, and blood samples were collected before and after GKS. NKA was measured using an enzyme-linked immunosorbent assay kit, to measure interferon-gamma (IFNγ) secreted by ex vivo-stimulated NK cells from whole blood. We explored the correlations between NK cell-produced IFNγ (NKA-IFNγ) levels and clinical parameters of patients who were treated with GKS for brain tumors. Results : NKA-IFNγ levels were decreased in metastatic brain tumor patients compared to those with benign brain tumors (p<0.0001). All the patients who used steroid treatment to reduce brain swelling after GKS had an NKA-IFNγ level of zero except one patient. High NKA-IFNγ levels were not associated with a rapid decrease in brain metastasis and did not increase after GKS. Conclusion : The activity of NK cells in metastatic brain tumors decreased more than that in benign brain tumors after GKS.

Enzymatic bioconversion of ginseng powder increases the content of minor ginsenosides and potentiates immunostimulatory activity

  • Park, Jisang;Kim, Ju;Ko, Eun-Sil;Jeong, Jong Hoon;Park, Cheol-Oh;Seo, Jeong Hun;Jang, Yong-Suk
    • Journal of Ginseng Research
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    • 제46권2호
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    • pp.304-314
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    • 2022
  • Background: Ginsenosides are biologically active components of ginseng and have various functions. In this study, we investigated the immunomodulatory activity of a ginseng product generated from ginseng powder (GP) via enzymatic bioconversion. This product, General Bio compound K-10 mg solution (GBCK10S), exhibited increased levels of minor ginsenosides, including ginsenoside-F1, compound K, and compound Y. Methods: The immunomodulatory properties of GBCK10S were confirmed using mice and a human natural killer (NK) cell line. We monitored the expression of molecules involved in immune responses via enzyme-linked immunosorbent assay, flow cytometry, NK cell-targeted cell destruction, quantitative reverse-transcription real-time polymerase chain reaction, and Western blot analyses. Results: Oral administration of GBCK10S significantly increased serum immunoglobulin M levels and primed splenocytes to express pro-inflammatory cytokines such as interleukin-6, tumor necrosis factor-α, and interferon-γ. Oral administration of GBCK10S also activated NK cells in mice. Furthermore, GBCK10S treatment stimulated a human NK cell line in vitro, thereby increasing granzyme B gene expression and activating STAT5. Conclusion: GBCK10S may have potent immunostimulatory properties and can activate immune responses mediated by B cells, Th1-type T cells, and NK cells.

Cholera Toxin B Subunit-Porphyromonas gingivalis Fimbrial Antigen Fusion Protein Production in Transgenic Potato

  • Lee, Jin-Yong;Kim, Mi-Young;Jeong, Dong-Keun;Yang, Moon-Sik;Kim, Tae-Geum
    • Journal of Plant Biotechnology
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    • 제36권3호
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    • pp.268-274
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    • 2009
  • Porphyromonas gingivalis, the gram-negative anaerobic oral bacterium, initiates periodontal disease by binding to saliva-coated oral surface. The cholera toxin B subunit (CTB) genetically linked to FimA1 (1-200 aa) or FimA2 (201-337 aa) of the P. gingivalis fimbrial antigen were introduced into Solanum tuberosum cells by Agrobacterium tumefaciens-mediated transformation method. The integration of CTB-FimA1 or CTB-FimA2 fusion genes were confirmed in the chromosome of transformed leaves by genomic DNA PCR amplification method. Synthesis and assembly of the CTB-FimA fusion proteins into oligomeric structures with pentamer size was detected in transformed tuber extracts by immunoblot analysis. The binding activities of CTB-FimA fusion proteins to intestinal epithelial cell membrane receptors were confirmed by GM1-ganglioside enzyme-linked immunosorbent assay (GM1-ELISA). The ELISA showed that the expression levels of the CTB-FimA1 or CTB-FimA2 fusion proteins were 0.0019, 0.002% of the total soluble protein in transgenic tuber tissues, respectively The synthesis of CTB-FimA monomers and their assembly into biologically active oligomers in transformed potato tuber tissues demonstrates the feasibility of using edible plants for the production of enterocyte targeted fimbrial antigens that could elicit mucosal immune responses.

실험적 Acanthamoeba 수막뇌염에서 세포성 면역에 관한 연구 (Cell-mediated immunity in experimental amoebic meningoencephalitis)

  • 임경일;정평림;김태우
    • Parasites, Hosts and Diseases
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    • 제27권2호
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    • pp.73-78
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    • 1989
  • 병원성이 강한 Acanthamoeba culbertsoni를 마우스에 감염시킨 후 경과된 감염기간에 따른 세포매개성 면역반응과 혈청 내 항체가의 변동을 관찰하였다. [9H]-thymidine을 이용한 마우스 비장세포의 아세포화를 관찰하였는데 사용된 mitogen으로는 concanavalin A(Con A)와 lipopoITsaccharide(LPS)였으며, 항체의 측정을 위하여 효소표식 면역검사법(ELISA)을 시행하였다. T림프구 및 B림프구의 아세포화 정도는 A. culbertsoni 감염 7일 후 증가하는 경향을 보였으며 다른 기간에 서는 대조군에 비해 별다른 차이를 보이지 않았다. 또한 감염된 마우스의 혈청 내 항체가도 감염 7 일 후부터 증가하는 경향을 관찰할 수 있었다.

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Induction of Apoptosis in Glioma Cells and Upregulation of Fas Expression Using the Human Interferon-β Gene

  • Guo, Yan;Wang, Gan;Gao, Wen-Wei;Cheng, Shi-Wen;Wang, Ren;Ju, Shi-Ming;Cao, He-Li;Tian, Heng-Li
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권6호
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    • pp.2837-2840
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    • 2012
  • We investigated whether IFN-${\beta}$ inhibits the growth of human malignant glioma and induces glioma cell apoptosis using the human IFN-${\beta}$ gene transfected into glioma cells. A eukaryonic expression vector ($pSV2IFN{\beta}$) for IFN-${\beta}$ was transfected into the glioma cell line SHG44 using liposome transfection. Stable transfection and IFN-${\beta}$ expression were confirmed using an enzyme-linked immunosorbent assay (ELISA). Cell apoptosis was also assessed by Hoechst staining and electron microscopy. In vivo experiments were used to establish a SHG44 glioma model in nude mice. Liposomes containing the human IFN-${\beta}$ gene were injected into the SHG44 glioma of nude mice to observe glioma growth and calculate tumor size. Fas expression was evaluated using immunohistochemistry. The IFN-${\beta}$ gene was successfully transfected and expressed in the SHG44 glioma cells in vitro. A significant difference in the number of apoptotic cells was observed between transfected and non-transfected cells. Glioma growth in nude mice was inhibited in vivo, with significant induction of apoptosis. Fas expression was also elevated. The IFN-${\beta}$ gene induces apoptosis in glioma cells, possibly through upregulation of Fas. The IFN-${\beta}$ gene modulation in the Fas pathway and apoptosis in glioma cells may be important for the treatment of gliomas.

HER2/neu Expression in Head and Neck Squamous Cell Carcinoma Patients is not Significantly Elevated

  • Sardari, Yasaman;Pardis, Soheil;Tadbir, Azadeh Andisheh;Ashraf, Mohammad Javad;Fattahi, Mohammad Javad;Ebrahimi, Hooman;Purshahidi, Sara;Khademi, Bijan;Hamzavi, Marzieh
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권6호
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    • pp.2891-2896
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    • 2012
  • Background: HER2/neu, a member of EGFR family, is over expressed in some tumors. The purpose of this study was to determine the salivary level and tissue expression of HER2/neu in patients with head and neck squamous cell carcinoma (HNSCC) and any correlation with clinicopathologic parameters. Methods: An enzyme-linked immunosorbent assay (ELISA) was used to evaluate the salivary level and immunohistochemistry (IHC) to assess tissue expression of HER2/neu in 28 patients with HNSCC and 25 healthy controls. Results: The salivary levels of HER2/neu in HNSCC patients was not significantly higher than in the healthy controls (p>0.005). There was no apparent correlation in salivary HER2/neu level with clinicopathological features such as age, sex, grade, tumor size and nodal status. All HNSCC specimens were positive (membranous or/and cytoplasmic) for HER2/neu, except one sample. Only one HNSCC specimen was stained in cytoplasm purely. All control specimens were membranous and cytoplasmic positive for HER2/neu. There was a significant difference between cytoplasmic staining in case and control groups (p-value<0.05). Conclusion: In our cases, no overexpression of HER2/neu was observed. Thus, our findings suggested that the use of Her-2 as a salivary marker of HNSCC cannot be recommended.

Salivary and Serum Interleukin-6 Levels in Oral Premalignant Disorders and Squamous Cell Carcinoma: Diagnostic Value and Clinicopathologic Correlations

  • Dineshkumar, Thayalan;Ashwini, Balakuntla Krishnamurthy;Rameshkumar, Annasamy;Rajashree, Padmanaban;Ramya, Ramadas;Rajkumar, Krishnan
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권11호
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    • pp.4899-4906
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    • 2016
  • Aim: To assess the diagnostic utility of serum and salivary interleukin 6 (IL-6) levels in the differential diagnosis of potentially malignant lesions and conditions (PMLs/PMCs) and oral squamous cell carcinoma (OSCC) in a high oral cancer prevalence region. Methods: After appropriate ethical clearance and informed consent, salivary and blood samples were collected from 100 participants in each group (OSCC, PMLs, and healthy controls). Serum and salivary IL-6 levels were measured by enzyme-linked immunosorbent assay and data were subjected to appropriate statistical analysis. Results: Significant differences in IL-6 concentration were noted between OSCC and PML/C patients in both serum and saliva, with salivary levels being 2 to 3 fold higher than serum values in all the groups. Receiver operating characteristic curve analysis demonstrated 96% specificity and 99% sensitivity for salivary IL-6 in differentiating PML from OSCC. Conclusions: The results of the present study suggest that the pro-inflammatory cytokine, IL-6, is elevated in the saliva of patients with OSSC compared to PMD and controls, and thus may prove to have diagnostic and/or prognostic significance.

Inhibition of Trypsin-Induced Mast Cell Activation by Water Fraction of Lonicera japonica

  • Kang, Ok-Hwa;Choi, Yeon-A;Park, Hye-Jung;Lee, Joo-Young;Kim, Dae-Ki;Choi, Suck-Chei;Kim, Tae-Hyun;Nah, Yong-Ho;Yun, Ki-Jung;Choi, Suck-Jun;Kim, Young-Ho;Bae, Ki-Hwan;Lee, Young-Ml
    • Archives of Pharmacal Research
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    • 제27권11호
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    • pp.1141-1146
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    • 2004
  • Lonicera japonica Thunb.(Caprifoliaceae) has long been known as an anti-inflammatory. In the present study, the effect of water fraction of Lonicera japonica (LJ) on trypsin-induced mast cell activation was examined. HMC-1 cells were stimulated with trypsin (100 nM) in the presence or absence of LJ (10, 100, and 1000 $\mu$ g/mL). TNF-$\alpha$ and tryptase production were measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-PCR. Extracellular signal-regulated kinase (ERK) phosphorylation was assessed by Western blot. Trypsin activity was measured by using Bz-DL-Arg-p-nitroanilide (BAPNA) as substrate. LJ (10, 100, and 1000 $\mu$g/mL) inhibited TNF-$\alpha$ secretion in a dose-dependent manner. LJ (10, 100, and 1000 $\mu$g/mL) also inhibited TNF-$\alpha$ and tryptase mRNA expression in trypsin-stimulated HMC-1. Furthermore, LJ inhibited trypsin-induced ERK phosphorylation. However, LJ did not affect the trypsin activity even 1000 $\mu$g/mL. These results indicate that LJ may inhibit trypsin-induced mast cell activation through the inhibition of ERK phosphorylation than the inhibition of trypsin activity.

Nypa fruticans wurmb regulates the secretion level of inflammatory cytokines in vitro models.

  • Jin, Yu-Mi;Kim, Seong-Seon;Lee, Jong-Hyun;Jeon, Yong-Deok;Jin, Jong-Sik
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2018년도 추계학술대회
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    • pp.128-128
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    • 2018
  • Nypa fruticans wurmb (NF) has been used as traditional medicinal food in Asian countries. Especially, NF has been used for conventional medicine to treat inflammatory periodontal diseases. Previous studies have been shown that NF has large amount of useful constituents such as phenolic acids, polyphenols and flavonoids. Also, NF is known as having medicinal effects such as anti-oxidant, anti-inflammatory and cholesterol-lowering effects. NF has recently been attracted to use complementary medicinal food on inflammatory diseases in Korea. However, there are no obvious effects in inflammatory and metabolic diseases also mechanisms has been studied yet. The purpose of this study was to investigate the anti-inflammatory effects of NF and steamed-NF (SNF), which recently has been used as health food, using Human keratinocyte cell line (HaCaT) and Human mast cell line (HMC-1). The cytotoxicities of NF and SNF were measured by using MTT assays in HaCaT cells and HMC-1 cell. To evaluate anti-inflammatory effects of NF and SNF, HaCaT cells were stimulated with tumor necrosis factor $(TNF)-{\alpha}$ and Interferon $(IFN)-{\gamma}$. Also, HMC-1 cells were stimulated with phorbol-12-myristate-13-acetate (PMA) and A23187 calcium ionophore (A23187) to induce allergic inflammation. Inflammatory cytokine were measured by enzyme-linked immunosorbent assay (ELISA). In this result, the extract of NF and SNF (0.01 - 1mg/ml) did not show cytotoxicity in HaCaT cells and HMC-1 cells. In addition, the NF and SNF suppressed the production of interleukin (IL)-6 and IL-8 in HaCaT cells at highest concentration. Furthermore, the treatment of SNF significantly inhibited the secretion level of IL-8 in PMA plus A23187-stimulated HMC-1 cells compared with NF treatment group. These results suggest that the extract of NF and SNF may serve as a potential therapy for skin inflammatory diseases.

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