• Journal of Microbiology and Biotechnology
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• v.19 no.2
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• pp.208-214
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• 2009

Steroid hormones have long been known to have a profound influence on the immune system. Although the functions of the nuclear receptors in the development of T cells are fairly well studied, the differential expression of these receptors in T helper cells is poorly understood. Here, we investigated the differential expression of nuclear receptors and coregulators in Th1 and Th2 cells by genome-wide micro array analysis. The result showed that several nuclear receptors and coregulators are differentially expressed in these cells. The result was confirmed by RT-PCR. The result showed that $RXR{\alpha}$ is highly expressed in Th2 cells. Overexpression of $RXR{\alpha}$ in a Jurkat human T cell line induced IL4 but not IFN-${\gamma}$ gene expression, suggesting that $RXR{\alpha}$ plays a selective role in Th1 and Th2 differentiation. In summary, these results suggest that Th1/Th2 differentiation is influenced by differential regulation of nuclear receptors and coregulators.

• KSBB Journal
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• v.26 no.3
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• pp.248-254
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• 2011

Various biometerials have been researched and have been developed for treatment of some disease through transplantation to body. They have been evaluated by in vitro cytotoxicity test using some skin-derived cell lines for prediction of their biocompatibility in vivo. However, the results of experiments using mesenchymal or epithelial cells could not be considered in vivo immune reaction. In this study, we evaluated the biomaterial-elution (elute from high density polyethylene film) using some cell lines (L929, Jurkat, U937) in vitro, and then that results were compared with in vivo results from guinea pig sensitization test. In sensitization test, saline and elution of syringe could not induce erythema, but only DNCB (hypersensitive chemical) induce erythema at guinea pig sensitization test. In cell experiment, the cytotoxicity results of inflammatory cells (Jurkat; T lymphocyte, U937; monocyte) was no difference with L929 (fibroblast) in the overall trend. However, inflammatory cell lines were only secreted inflammatory cytokine (TNF-${\alpha}$, INF-${\gamma}$) in some materials (biomateriallution, FAC, DNCB). And the biomaterial-elution did not have toxicity to the cells, but it induced the inflammatory cytokines in inflammatory cell lines only. So, we were predicted inflammatory reaction through the cytokine resultes of inflammatory cell lines, and it was more correlated with in vivo results than cytotoxicity test. Therefore, we suggested that the inflammatory cytokine assay using inflammatory cell lines are more effective method in vitro for evaluation of biocompatibility of biomaterials or chemicals.

• Journal of Radiation Protection and Research
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• v.43 no.2
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• pp.66-74
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• 2018

Background: Tumor response to anticancer therapies can much be influenced by microenvironmental factors. In this study, we determined the effect of these microenvironmental factors on DNA methylation using A549 human lung adenocarcinoma cell line. Materials and Methods: We subjected A549 cells to various conditions mimicking tumor microenvironment including hypoxia, acidosis (sodium lactate), oxidative stress ($H_2O_2$), bystander effect (supernatant from doxorubicin (Dox)-treated or irradiated cells), and immune cell infiltration (supernatant from THP-1 or Jurkat T cells). Genomic DNA was isolated from these cells and analyzed for DNA methylation. Clonogenic cell survival, gene expression, and metabolism were analyzed in cells treated with some of these conditions. Results and Discussion: We found that DNA methylation level was significantly decreased in A549 cells treated with conditioned media from Dox-treated cells or Jurkat T cells, or sodium lactate, indicating an active transcription. To determine whether the decreased DNA methylation affects radiation sensitivity, we exposed cells to these conditions followed by 6 Gy irradiation and found that cell survival was significantly increased by sodium lactate while it was decreased by conditioned media from Dox-treated cells. We further observed that cells treated with conditioned media from Dox-treated cells exhibited significant changes in expression of genes including BAX and FAS (involved in apoptosis), NADPH dehydrogenase (mitochondria), EGFR (cellular survival) and RAD51 (DNA damage repair) while sodium lactate increased cellular metabolism rather than changing the gene expression. Conclusion: Our results suggest that various tumor microenvironmental factors can differentially influence DNA methylation and hence radiosensitivity and gene expression in A549 cancer cells.

• Journal of Microbiology and Biotechnology
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• v.6 no.6
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• pp.474-481
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• 1996

Since the commercially available rabbit anti-cyclin D3, generated from c-terminal 16 amino acid residues which are common to human and murine cyclin D3, is highly cross-reactive with many other cellular proteins of mouse, a new rabbit polyclonal anti-cyclin D3 has been raised by using murine cyclin D3 protein expressed at a high level in Escherichia coli as the immunogen. To express murine cyclin D3 protein in E. coli, the cyclin D3 cDNA fragment encoding c-terminal 236 amino acid residues obtained by polymerase chain reaction (PCR) was inserted into the NcoI/BamHI site of protein expression vector, pET 3d. Molecular mass of the cyclin D3 overexpressed in the presence of IPTG (Isopropyl $\beta$-D-thiogalactopyranoside) was approximately 26 kDa as calculated from the reading frame on the DNA sequence, and the protein was insoluble and mainly localized in the inclusion bodies that could be easily purified from the other cellular soluble proteins. When renaturation was performed following denaturation of the insoluble cyclin D3 protein in the inclusion bodies using guanidine hydrochloride, 4.4 mg of soluble form of cyclin D3 protein was produced from the transformant cultured in 100ml of LB media under the optimum conditions. Four-hundred micrograms of the soluble form of cyclin D3 protein was used for each immunization of a rabbit. When the antiserum obtained 2 weeks after tertiary immunization was applied to Western blot analysis, it was able to detect 33 kDa cyclin D3 protein in both murine lymphoma cell line BW5147.G.1.4 and human Jurkat T cells at 3,000-fold dilution with higher specificity to murine cyclin D3, demonstrating that the new rabbit polyclonal anti-murine cyclin D3 generated against c-terminal 236 amino acid residues more specifically recognizes murine cyclin D3 protein than does the commercially available rabbit polyclonal antibody raised against c-terminal 16 amino acids residues.

• Korean Journal of Medicinal Crop Science
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• v.11 no.1
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• pp.62-70
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• 2003

The biological activities on human immune and cancer cell lines of the four kinds of Korean mistletoes (Korean Viscum album, var. coloratum, : Korean Viscum sp. in Quercus acutissima Carr., Korean Viscum sp. in Castanea crenata, Korean Viscum sp. in Betula platyphylla, and Korean Viscum sp. in Salix koreensis) extracts were investigated. The extracts were preparated with ethanol, and fractionated with n-butanol, ethyl acetate, chloroform, hexane, and second distilled water. Cytotoxic potencies of the fractions on human normal lung cell line (HEL 299) showed under 28% in the concentration of 0.5 mg/ml. Growth inhibition effect of the Korean mistletoe extracts on the several human cancer cell lines depends on the concentration of the extracts, and extracting solvent. The hexane, chloroform, and ethyl acetate fractions indicated a strong anticancer activity, but not in aqueous and butanol fractions. Some mistletoe fractions have a different characteristic on the cancer cell lines. Stimulation on the growth of human immuno cell lines(B cell : Raji, T cell: Jurkat) of the extracts were confirmed in the ethyl acetate, chloroform, hexane fractions, but not in aqueous system.