• Biomolecules & Therapeutics
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• v.4 no.1
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• pp.46-50
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• 1996

Inhibition of the growth of human cancer cells by proteins secreted from 373-L1 cells was investigated in the present study. The growth of human cancer cells was inhibited by co-culture with 373-L1 cells under 10% FBS and DME, DME, GIT and serumless medium, respectively. The conditioned medium of cultured 373-L1 cells under serumless medium was concentrated 100-fold through an ultrafiltration cell with a 10,000 molecular weight cutoff at 4$^{\circ}C$ under positive pressure using nitrogen(373-L1 EM). 373-L1 EM inhibited the growth of HeLa, Hep G 2, KHOS-Np, A43l and MCF-7 cells. 3T3-L1 EM was purified with FPLC, DEAE-ion exchange chromatography and phenyl-sepharose chromatography. The major protein of 373-L1 EM has a molecular weight of 66,000-68,000 in SDS-PAGE analysis. The results suggest that the inhibitory activity of 373-L1 EM appears to be due to some protein(m.w.66,000-68,000) secreted by 373-L1 cells.

• IMMUNE NETWORK
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• v.23 no.2
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• pp.17.1-17.16
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• 2023

Latent membrane protein 2A (LMP2A), a latent Ag commonly expressed in Epstein-Barr virus (EBV)-infected host cells, is a target for adoptive T cell therapy in EBV-associated malignancies. To define whether individual human leukocyte antigen (HLA) allotypes are used preferentially in EBV-specific T lymphocyte responses, LMP2A-specific CD8⁺ and CD4⁺ T cell responses in 50 healthy donors were analyzed by ELISPOT assay using artificial Ag-presenting cells expressing a single allotype. CD8⁺ T cell responses were significantly higher than CD4⁺ T cell responses. CD8⁺ T cell responses were ranked from highest to lowest in the order HLA-A, HLA-B, and HLA-C loci, and CD4⁺ T cell responses were ranked in the order HLA-DR, HLA-DP, and HLA-DQ loci. Among the 32 HLA class I and 56 HLA class II allotypes, 6 HLA-A, 7 HLA-B, 5 HLA-C, 10 HLA-DR, 2 HLA-DQ, and 2 HLA-DP allotypes showed T cell responses higher than 50 spot-forming cells (SFCs)/5×105 CD8⁺ or CD4⁺ T cells. Twenty-nine donors (58%) showed a high T cell response to at least one allotype of HLA class I or class II, and 4 donors (8%) had a high response to both HLA class I and class II allotypes. Interestingly, we observed an inverse correlation between the proportion of LMP2A-specific T cell responses and the frequency of HLA class I and II allotypes. These data demonstrate the allele dominance of LMP2A-specific T cell responses among HLA allotypes and their intra-individual dominance in response to only a few allotypes in an individual, which may provide useful information for genetic, pathogenic, and immunotherapeutic approaches to EBV-associated diseases.

• Molecules and Cells
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• v.47 no.2
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• pp.100031.1-100031.13
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• 2024

It is now well-accepted that obesity-induced inflammation plays an important role in the development of insulin resistance and type 2 diabetes. A key source of the inflammation is the murine epididymal and human visceral adipose tissue. The current paradigm is that obesity activates multiple proinflammatory immune cell types in adipose tissue, including adipose-tissue macrophages (ATMs), T Helper 1 (Th1) T cells, and natural killer (NK) cells, while concomitantly suppressing anti-inflammatory immune cells such as T Helper 2 (Th2) T cells and regulatory T cells (Tregs). A key feature of the current paradigm is that obesity induces the anti-inflammatory M2 ATMs in lean adipose tissue to polarize into proinflammatory M1 ATMs. However, recent single-cell transcriptomics studies suggest that the story is much more complex. Here we describe the single-cell genomics technologies that have been developed recently and the emerging results from studies using these technologies. While further studies are needed, it is clear that ATMs are highly heterogeneous. Moreover, while a variety of ATM clusters with quite distinct features have been found to be expanded by obesity, none truly resemble classical M1 ATMs. It is likely that single-cell transcriptomics technology will further revolutionize the field, thereby promoting our understanding of ATMs, adipose-tissue inflammation, and insulin resistance and accelerating the development of therapies for type 2 diabetes.

• Journal of Life Science
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• v.16 no.4
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• pp.589-597
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• 2006

Genistein, a natural isoflavonoid phytoestrogen, is a strong inhibitor of protein tyrosine kinase and DNA topoisomerase activities. There are several studies documenting molecular alterations leading to cell cycle arrest and induction of apoptosis by genistein as a chemopreventive agent in a variety of cancer cell lines; however, its mechanism of action and its molecular targets on human bladder carcinoma and leukemic cells remain unclear. In the present study, we have addressed the mechanism of action by which genistein suppressed the proliferation of T24 bladder carcinoma and U937 leukemic cells. Genistein significantly inhibited the cell growth and induced morphological changes, and induced the G2/M arrest of the cell cycle in both T24 and U937 cells with a relatively stronger cytotoxicity in U937. The G2/M arrest in T24 cells was associated with the inhibition of cyclin A, cyclin B1 and Cdc25C protein expression without alteration of tumor suppressor p53 and cyclin-dependent kinase (Cdk) inhibitor p21(WAF1/CIP1). However, the inhibitory effects of genistein on the cell growth of U937 cells were connected with a marked inhibition of cyclin B1 and an induction of Cdk inhibitor p21 proteins by p53-independent manner. These data suggest that genistein may exert a strong anticancer effect and additional studies will be needed to evaluate the different mechanisms between T24 and U937 cells.

• Korean Journal of Medicinal Crop Science
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• v.10 no.1
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• pp.51-57
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• 2002

The biological activities of immune modulating activities of the extracts from Echinacea purpurea, Chrysanthmum indicum L. and Circium japonicum var. ussuriense KITAMURA were compared. About 70% of the growth of human hepatocarcinoma and 80% of human gastric cancer cell was inhibited in adding 0.5mg/ml of the ethanol extracts of Echinacea purpurea, Chrysanthmum indicum L. and Circium japonicum var. ussuriense KITAMURA, respectively. The growth of human breast cancer cells was also inhibited in adding 0.5mg/ml of the extracts as well as 60% of the human cancer cells. It was proved that the growth of human normal lung cell, scored as 15% for the extracts. Overall selectivity of the extracts on several human cancer cell line was over 3, which is higher than those from the conventional herbs. The growth of both human immune B and T cells was enhanced up to 1.4 to 2.0 times by adding the extracts, compared to the controls. The secretion of tumor necrosis $factor-alpha(TNF-{\alpha})$ from T cell was also increased up to 94 pg/ml in adding the Echinacea purpurea ethanol extract (0.5mg/ml). Circium japonicum var. ethanol extract also increased up to about 96 pg/ml of interleukin-6(IL-6) from B cell.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.121-121
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• 2003

The standard protocol for the production of transgenic mouse from ES-injected embryo has to process via chimera producing and several times breeding steps, In contrast, tetraploid-ES cell complementation method allows the immediate generation of targeted murine mutants from genetically modified ES cell clones. The advantage of this advanced technique is a simple and efficient without chimeric intermediates. Recently, this method has been significantly improved through the discovery that ES cells derived from hybrid strains support the development of viable ES mice more efficiently than inbred ES cells do. Therefore, the objective of this study was to generate transgenic mice overexpressing human resistin gene by using tetrapioid-ES cell complementation method. Human resistin gene was amplified from human fetal liver cDNA library by PCR and cloned into pCR 2.1 TOPO T-vector and constructed in pCMV-Tag4C vector. Human resistin mammalian expression plasmid was transfected into D3-GL ES cells by lipofectamine 2000, and then after 8~10 days of transfection, the human resistin-expressing cells were selected with G418. In order to produce tetraploid embryos, blastomeres of diploid embryos at the two-cell stage were fused with two times of electric pulse using 60 V 30 $\mu$sec. (fusion rate : 93.5%) and cultured upto the blastocyst stage (development rate : 94.6%). The 15~20 previously G418-selected ES cells were injected into tetraploid blastocysts, and then transferred into the uterus of E2.5d pseudopregnant recipient mice. To investigate the gestation progress, two El9.5d fetus were recovered by Casarean section and one fetus was confirmed to contain human resistin gene by genomic DNA-PCR. Therefore, this finding demonstrates that tetraploid-ES mouse technology can be considered as a useful tool to produce transgenic mouse for the rapid analysis of gene function in vivo.

• Journal of Nutrition and Health
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• v.43 no.2
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• pp.152-160
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• 2010

The purpose of this study was to investigate the effects of aging process on the immunity in human subjects. In this investigation, nineteen families of three generations (daughters on college age, their mothers, and grandmothers) participated to avoid genetic variation among individuals. Dietary food records, anthropometric measurements and biochemical assessments of serum nutrients were used to evaluate the nutritional status of subjects. The immune parameters of subjects were assessed by the total and differential WBC count. Total B and T lymphocytes, and T cell subsets were quantified by flowcytometer. Serum immunoglobulin G, A, M concentrations were also measured as an index of humoral immunity. The result of this study can be summarized as follows: 1. Along with the aging process, body fat was found to be increased whereas lean body mass and total body water were diminished. Since there were no significant difference in serum vitamin E levels in all age groups, serum retinal concentrations tended to decrease as one gets old. 2. Although total number of T lymphocytes seemed to be unchanged, B lymphocytes and NK cell numbers were increased by aging. The Percentage of CD8 + lymphocytes was lower in the elderly subjects compared with the younger, resulting in higher ratio of CD4 +/CD8 + lymphocytes in the elderly. Serum Ig G and Ig A levels remained unchanged, but IgM levels were significantly decreased as the age processes continue. Taking all together, it could be suggested that the alteration of immune cell population by aging is selective and possibly nonage factors such as nutrition may be attributable to the change of immunity in the elderly. The nutritional status and aging process may selectively affect both the cell-mediated (CD8 +, CD4 + CD8 + ratio, NK cell) and humoral (B lymphocyte, Immunoglobulin M, G) immune parameters in human subjects.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.3
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• pp.367-373
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• 2014

In this study, we investigated the anticancer activity of methyl gallate (MG), which is the major biologically active component of Galla Rhois, in RC-58T/h/SA#4 human prostate cancer cells. MG inhibited cell proliferation in a dose-dependent manner. Cell death induced by MG increased the population of cells in sub-G1 phase, formation of apoptotic bodies, nuclear condensation, and DNA fragmentation. Apoptosis induced by MG was associated with activation of initiator caspases-8 and -9 as well as effector caspase-3. Endocrine disruptors such as dioxin and bisphenol A increased growth of RC-58T/h/SA#4 cells in charcoal-treated FBS (cFBS) medium. Cell proliferation was highest upon treatment with 1 nM and $0.1{\mu}M$ dioxin and bisphenol A, respectively. MG also dose-dependently inhibited cell proliferation in RC-58T/h/SA#4 cells treated with endocrine disruptors. These results indicate that MG exerts anticancer effects on RC-58T/h/SA#4 primary human prostate cancer cells.

• The Korean Journal of Physiology and Pharmacology
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• v.25 no.3
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• pp.251-258
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• 2021

Flos magnoliae (FM), the dry flower buds of Magnolia officinalis or its related species, is a traditional herbal medicine commonly used in Asia for symptomatic relief of and treating allergic rhinitis, headache, and sinusitis. Although several studies have reported the effects of FM on store-operated calcium entry (SOCE) via the ORAI1 channel, which is essential during intracellular calcium signaling cascade generation for T cell activation and mast cell degranulation, the effects of its isolated constituents on SOCE remain unidentified. Therefore, we investigated which of the five major constituents of 30% ethanoic FM (vanillic acid, tiliroside, eudesmin, magnolin, and fargesin) inhibit SOCE and their physiological effects on immune cells. The conventional whole-cell patch clamp results showed that fargesin, magnolin, and eudesmin significantly inhibited SOCE and thus human primary CD4+ T lymphocyte proliferation, as well as allergen-induced histamine release in mast cells. Among them, fargesin demonstrated the most potent inhibitory effects not only on ORAI1 (IC50 = 12.46 ± 1.300 μM) but also on T-cell proliferation (by 87.74% ± 1.835%) and mast cell degranulation (by 20.11% ± 5.366%) at 100 μM. Our findings suggest that fargesin can be a promising candidate for the development of therapeutic drugs to treat allergic diseases.

• Journal of Life Science
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• v.17 no.12
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• pp.1641-1648
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• 2007

Yippee-like proteins, which have been identified as the homolog of Drosophila yippee protein containing a zinc-finger domain, are known to be highly conserved among eukaryotes. However, their functional roles are still poorly understood. Recently we initiated ordered differential display (ODD)-polymerase chain reaction (PCR) to isolate genes of which expressions are altered following activation of human T cells. On the ODD-PCR image, one PCR-product detected in unstimulated T cells was not detectable at the time when the activated T cells traversed near $G_1/S$ boundary following activation by immobilized anti-CD3. Cloning and nucleotide sequence analysis revealed that the PCR-product was yippee-like 5 (YPEL5) gene, which was known as a human homolog of the Drosophila yippee gene. Northern blot analysis confirmed the amount of ${\sim}2.2$ kb YPEL5 mRNA expression detectable in unstimulated T cells was sustained until 1.5 hr after activation and then rapidly declined to undetectable level by 5 hr. Ectopic expression of YPEL5 gene in human cervix epitheloid carcinoma HeLa cells caused a significant reduction in cell proliferation to the level of 47% of the control. Expression of GFP-YPEL5 fusion protein in HeLa cells showed its nuclear localization. These results demonstrated that the expression level of human YPEL5 mRNA was negatively regulated in the early stage of T cell activation, and suggested that YPEL5 might exert an inhibitory effect on the cell proliferation as a nuclear protein.