• Title/Summary/Keyword: mononuclear cells

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Heat-Shock Protein 70 as a Tumor Antigen for in vitro Dendritic Cell Pulsing in Renal Cell Carcinoma Cases

  • Meng, Fan-Dong;Sui, Cheng-Guang;Tian, Xin;Li, Yan;Yang, Chun-Ming;Ma, Ping;Liu, Yun-Peng;Jiang, You-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8947-8950
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    • 2014
  • Immunological functions of heat shock proteins (HSPs) have long been recognized. In this study we aimed to efficiently purify HSP70 from renal cell carcinoma and test it as a tumor antigen for pulsing dendritic cells in vitro. HSP70 was purified from renal cell carcinoma specimens by serial column chromatography on Con A-sepharose, PD-10, ADP-agarose and DEAE-cellulose, and finally subjected to fast protein liquid chromatography (FPLC). Dendritic cells derived from the adherent fraction of peripheral blood mononuclear cells were cultured in the presence of IL-4 and GM-CSF and exposed to tumor HSP70. After 24 hours, dendritic cells were phenotypically characterized by flow cytometry. T cells obtained from the non-adherent fraction of peripheral blood mononuclear cells were then co-cultured with HSP70-pulsed dendritic cells and after 3 days T cell cytotoxicity towards primary cultured renal cell carcinoma cells was examined by Cell Counting Kit-8 assay. Dendritic cells pulsed in vitro with tumor-derived HSP70 expressed higher levels of CD83, CD80, CD86 and HLA-DR maturation markers than those pulsed with tumor cell lysate and comparable to that of dendritic cells pulsed with tumor cell lysate plus TNF-${\alpha}$. Concomitantly, cytotoxic T-lymphocytes induced by HSP70-pulsed dendritic cells presented the highest cytotoxic activity. There were no significant differences when using homologous or autologous HSP70 as the tumor antigen. HSP70 can be efficiently purified by chromatography and induces in vitro dendritic cell maturation in the absence of TNF-${\alpha}$. Conspecific HSP70 may effectively be used as a tumor antigen to pulse dendritic cells in vitro.

Regulatory Mechanism of Lipopolysaccharide(LPS)-Induced Interleukin-8 Gene Expression in Mononuclear Phagocytic Cells (단핵식세포에서 내독소에 의한 인터루킨-8 유전자 발현 조절기전에 관한 연구)

  • Yoo, Chul-Gyu;Suh, Gee-Young;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.5
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    • pp.462-474
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    • 1994
  • Background : In acute lung injury, activated neutrophils play an important role in tissue damage. For neutrophils to participate in lung inflammation, chemotactic factors released from mononuclear phagocytes are needed to bring these cells to the local site of inflammation, with interleukin-8 (IL-8) being one of the most specific and important chemotactic factors for neutrophils. IL-8 also induces the expression of adhesion molecules and activates neutrophils to release various inflammatory mediators. Lipopolysaccharide(LPS) is one of the most important causes of adult respiratory distress syndrome and can cause release of many inflammatory cytokines including IL-8 leading to acute lung injury. But little is known about the regulatory mechanism of LPS-induced IL-8 gene expression in mononuclear phagocytes. Method : Human alveolar macrophages(HAM) and peripleral blood monocytes(PBMC) were isolated from healthy volunteers. Time and dose relationship of LPS-induced IL-8 mRNA expression was observed by Northern blot analysis. To evaluate the regulatory mechanism of LPS-induced IL-8 gene expression, pretreatment of actinomycin D(AD, $5{\mu}g/ml$) and cycloheximide(CHX, $5{\mu}g/ml$) was done and Northern blot analysis for IL-8 mRNA and ELISA for immunoreactive IL-8 protein in culture supernatant were performed. Results : 1) In HAM, dose and time dependent LPS-induced IL-8 mRNA expression was observed with peak mRNA level at 8 hours post-stimulation. 2) In PBMC, dose and time dependent LPS-induced IL-8 mRNA expression was also observed with peak mRNA level at 4 hours post-stimulation. 3) AD decreased expression of LPS-induced IL-8 gene expression at both mRNAand protein levels in both types of cells. 4) CHX decreased expression of LPS-induced IL-8 gene expression at protein level in both cell types but in HAM, superinduction of IL-8 mRNA was observed while decreased expression of IL-8 mRNA was observed in PBMC. Conclusion : Time and dose dependent LPS-induced IL-8 gene expression was observed in mononuclear phagocytes which is at least partly regulated pretranslationally. LPS-induced IL-8 mRNA expression in HAM needs no de novo protein synthesis and may be under the control of a labile repressor protein while de novo protein synthesis may be needed in PBMC.

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Effect of Egg White Derivatives on Phagocytic Response of Feline Peripheral Blood Phagocytes (고양이 말초혈액 탐식세포의 탐식능에 있어서 계난백유래물질의 효과)

  • 양만표;김기홍
    • Journal of Veterinary Clinics
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    • v.16 no.1
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    • pp.37-41
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    • 1999
  • The immunostimulating effects of egg white derivatives (EWD) on the phagocytic response of feline peripheral blood polymorphonuclear cells (PMN) as well as mono- nuclear cells (MNC) were examined. The phagocytic activity was analyzed by a flow cytometry system. The EWD showed directly an enhanced effect on the phagocytic response of MNC but not PMN. The phagocytic activity of MNC was enhanced by culture supernatant from MNC and PMN treated with EWD, respectively. Similarly, the phagocytic activity of PMN was enhanced by culture supernatant from MNC but not PMN treated with EWD. It was, therefore, indicated that the enhanced phagocytic activity of feline PMN could be mainly mediated by humoral factor(s) released from MNC treated with EWD. These results suggested that EWD could enhance the phagocytosis of feline peripheral blood phagocytes.

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In Vitro Selection of Cancer-Specific RNA Aptamers

  • Lee Young-Ju;Lee Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1149-1153
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    • 2006
  • In this study, nuclease-resistant RNA aptamers that are specific for Jurkat T leukemia cells were selected by a subtractive systemic evolution of ligands by exponential enrichment (SELEX) method. A randomized nuclease-resistant RNA library was incubated with normal peripheral blood mononuclear cells (PBMC) in each round to preclude RNAs that recognize the common cellular components on the surface of normal and cancer cells. The precluded RNAs were used for the selection of Jurkat T cell-specific aptamers, and the specific RNAs were then gradually enriched from start to the following selections. After 16 rounds of the subtractive SELEX, the selected aptamers were found to preferentially bind to Jurkat T cells, but not to the normal PBMC, evidenced by fluorescence-activated cell sorting analysis. Thus, the subtractive SELEX can be used to identify ligands to cancer-specific biological markers without prior knowledge of the nature of markers. The aptamers could be applied to specific cell sorting, tumor therapy, and diagnosis, and moreover, to find cancer cell-specific markers.

A STUDY OF THE MALIGNANT FIBROUS HISTIOCYTOMA (악성 섬유성 조직구종에 관한 연구)

  • Kim Young-Jin;Oh Phill-Gyo;Choi Eui-Hwan
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.24 no.2
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    • pp.451-457
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    • 1994
  • Malignant fibrous histiocytoma(MFH) is relatively rare in the oro-maxillofacial region, particularly in the oral cavity. MFH usually arise in the extremities, the thigh being the most common site. The incidence of malignant fibrous histiocytoma in bones is rather low compared with that in soft tissues. MFH is predominant in the 40s and 50s. Histologically, the lesion are said to show high cellularity with fibrous stroma, cellular and nuclear pleomorphism, an admixture of fibroblast-like spindle cells which tend to be arranged in whorls or cartwheel or storiform patterns, rounded mononuclear cells and multinucleated giant cells. The cells frequently have abundant eosinophilic cytoplasm which has a foamy or vesicular appearance. Treatment consists of varying combinations of radiation therapy, chemotherapy, and surgery. We have observed a case of malignant fibrous histiocytoma occured in the right maxilla of 32-year-old woman.

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Effects of Antioxidant Supplementation on Antioxidant Status and PHA-Stimulated Interleukin-2 Production by Peripheral Blood Mononuclear Cells in the Elderly Women

  • Kim Jung Hee;Kim Ok Hyon;Lim Jae Yeon
    • Journal of Community Nutrition
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    • v.7 no.1
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    • pp.42-48
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    • 2005
  • This study was done to investigate effects of antioxidant supplementation on phytohemagglutinin (PHA) -stimulated interleukin-2 (IL-2) production by peripheral blood mononuclear cells (PBMCs) in elderly women. This study was designed as a placebo-controlled, single-blinded, randomized intervention trial. Twenty four elderly women aged over 60 years, visitings social welfare center in Seoul were divided into 3 groups, placebo (n = 8), vitamin C supplemented (n = 8) , and vitamin E supplemented (n = 8) groups. Experimental groups were given either 1000mg of L-ascorbic acid or 400 ill of d- $\alpha$-tocopherol for 4 weeks. There was no significant difference in antioxidant vitamins intakes and their plasma levels among pre-intervention groups. Plasma vitamin C or E levels was significantly increased after vitamin C or E sup-plementations. The increases of plasma thiobarbituric acid-reactive substance (TBARS) levels in the placebo group were significantly higher than those of the supplemented 2 groups. There were no significant differences in the changes of plasma IL-2 level between pre- and post-intervention among the 3 groups. However there was a significant increase in PHA­stimulated IL-2 production by PBMCs after 4-week vitamin E or vitamin C supplementation. Particularly, vitamin E supplemented group showed a higher PHA-stimulated IL-2 production than vitamin C supplemented group. These results indicate that vitamin E or vitamin C supplementation might enhance mitogen-stimulated cytokine production by immune cells, which could be one of the factors to improve health status in the elderly.

Label-free Noninvasive Characterization of Osteoclast Differentiation Using Raman Spectroscopy Coupled with Multivariate Analysis

  • Jung, Gyeong Bok;Kang, In Soon;Lee, Young Ju;Kim, Dohyun;Park, Hun-Kuk;Lee, Gi-Ja;Kim, Chaekyun
    • Current Optics and Photonics
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    • v.1 no.4
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    • pp.412-420
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    • 2017
  • Multinucleated bone resorptive osteoclasts differentiate from bone marrow-derived monocyte/macrophage precursor cells. During osteoclast differentiation, mononuclear pre-osteoclasts change their morphology and biochemical characteristics. In this study, Raman spectroscopy with multivariate techniques such as Principal Component Analysis (PCA) and Linear Discriminant Analysis (LDA) were used to extract biochemical information related to various cellular events during osteoclastogenesis. This technique allowed for label-free and noninvasive monitoring of differentiating cells, and clearly discriminated four different time points during osteoclast differentiation. The Raman band intensity showed significant time-dependent changes that increased up to day 4. The results of Raman spectroscopy agreed with results from atomic force microscopy (AFM) and tartrate-resistant acid phosphatase (TRAP) staining, a conventional biological assay. Under AFM, normal spindle-like mononuclear pre-osteoclasts became round and smaller at day 2 after treatment with a receptor activator of nuclear $factor-{\kappa}B$ ligand and they formed multinucleated giant cells at day 4. Thus, Raman spectroscopy, in combination with PCA-LDA, may be useful for noninvasive label-free quality assessment of cell status during osteoclast differentiation, enabling more efficient optimization of the bioprocesses.

Cell Cycle Regulation and Induction of Apoptosis by β-carotene in U937 and HL-60 Leukemia Cells

  • Upadhyaya, K.R.;Radha, K.S.;Madhyastha, H.K.
    • BMB Reports
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    • v.40 no.6
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    • pp.1009-1015
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    • 2007
  • In this communication, we report the efficacy of $\beta$-carotene towards differentiation and apoptosis of leukemia cells. Dose ($20{\mu}M$) and time dependence (12 h) tests of $\beta$-carotene showed a higher magnitude of decrease (significance p < 0.05) in cell numbers and cell viability in HL-60 cells than U937 cells but not normal cell like Peripheral blood mononuclear cell (PBMC). Microscopical observation of $\beta$-carotene treated cells showed a distinct pattern of morphological abnormalities with inclusion of apoptotic bodies in both leukemia cell lines. When cells were treated with $20{\mu}M$ of $\beta$-carotene, total genomic DNA showed a fragmentation pattern and this pattern was clear in HL-60 than U937 cells. Both the cell lines, on treatment with $\beta$-carotene, showed a clear shift in $G_1$ phase of the cell cycle. In addition the study also revealed anti-oxidant properties of $\beta$-carotene since there was reduction in relative fluorescent when treated than the control at lower concentration. Collectively this study shows the dual phenomenon of apoptosis and differentiation of leukemia cells on treatment with $\beta$-carotene.

IL-23 P19 Expression Induced by IL-17 and $IL-1{\beta}$ in Rheumatoid Arthritis Synovial Mononuclear Cells (류마티스관절염 환자의 활액 세포에서 IL-17과 $IL-1{\beta}$에 의한 IL-23p19의 발현 증가)

  • Cho, Mi-La;Heo, Yu-Jung;Oh, Hye-Jwa;Kang, Chang-Min;Lee, Seon-Yeong;Hong, Yeon-Sik;Kim, Ho-Youn
    • IMMUNE NETWORK
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    • v.8 no.1
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    • pp.29-37
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    • 2008
  • Interleukin-23 (IL-23) is a novel pro-inflammatory cytokine which has been implicated to play a pathogenic role in rheumatoid arthritis (RA). This study was undertaken to investigate the IL-23 inductive activity of the proinflammatory cytokine IL-17, $IL-1{\beta}$ and tumor necrosis factor (TNF-${\alpha}$) in RA synovial fluid mononuclear cells (SFMC). Expression of IL-23p19, IL-17, $IL-1{\beta}$ and TNF-${\alpha}$ in joint was examined by immunohistochemistry (IHC) of patients with RA and osteoarthritis (OA). The effects of IL-17 and $IL-1{\beta}$ on expression of IL-23p19 in human SFMC from RA patients were determined by reverse transcriptase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). IL-23p19 was expressed in the RA fibroblast like synoviocyte (FLS), but not from OA FLS. Similar to the protein expression, IL-23p19 mRNA could be detected by RT-PCR in RA SFMC. IL-17 and $IL-1{\beta}$ could induce RA SFMC to produce the IL-23p19. The effects of IL-17 were much stronger than $IL-1{\beta}$ or TNF-${\alpha}$. These responses were observed in a doseresponsive manner. In addition, IL-17 or $IL-1{\beta}$ neutralizing antibody down-regulated the expression of IL-23p19 induced by LPS in RA-SFMC. Our results demonstrate that IL-23p19 is overexpressed in RA synovium and IL-17 and $IL-1{\beta}$ appears to upregulate the expression of IL-23p19 in RA-SFMC.

The Relationship between the Serum Cytokine and Clinical Improvement in Major Depressive Disorder (주요 우울증에서 혈중 Cytokine과 임상적 호전과의 관계)

  • Kim, Hyon Chul;Lee, Sang Kyu;Kim, Do Hoon;Son, Bong Ki
    • Korean Journal of Biological Psychiatry
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    • v.10 no.1
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    • pp.70-79
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    • 2003
  • Object : Currently, the alteration of cytokine system has been known to play an important role in regard to depressive symptom. We focused on the relationship between immunological parameters and clinical improvement in major depressive disorder. Method : Data were collected on 26 patients with major depressive disorder using a 8-week prospective follow-up design. After 8-week treatment period with fluoxetine, patients were classified into a response group and a non-response group according to their psychopathological outcome as evaluated by Hamilton Depression Rating Scale. The differences of the immunological parameters between pre-treatment phase and post-treatment phase were compared among patients. The difference of those was also compared within each phase among them. The relationship between socio-demographic variables, depression, cytokine, mononuclear cells was examined by correlation analysis. Multiple regression analyses were performed to explore the predictors of clinical improvement of major depressive disorder. Result : Pre-treatment levels of IL-$1{\beta}$ in the response group were significantly higher than those in the non-response group. Pre-treatment levels of IL-$1{\beta}$ of all patients and in the response group were positively correlated with pre-treatment monocyte counts. Patients with subsequent remission showed significantly lower IL-6 values at baseline than those with non-response. Post-treatment values of IL-6 did not differ significantly among the patients. The correlation test showed more frequent relations among cytokines and mononuclear cells in the response group than in the non-responder group. Especially, serum level of IL-6 in pre-treatment phase was only significantly correlated with HAMD score after 8-week treatement phase, while other cytokines and mononuclear cells were not. Pretreatment level of IL-6 was of paramount importance in predicting clinical improvement of depressive symptom. Conclusion : The immune system of major depressive disorder patients might dichotomize the patients into subsequent responders and non-responders. Immune system might be of great influence on the clinical improvement of major depressive disorder. The mode of interaction between depression and cellular immune function and the mediators responsible for the cytokine production need to be studied further.

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