• Title/Summary/Keyword: minCD

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Proteomic Analysis of Resting and Activated Human $CD8^+$ T Cells

  • Koo Jung-Hui;Chae Wook-Jun;Choi Je-Min;Nam Hyung-Wook;Morio Tomohiro;Kim Yu-Sam;Jang Yang-Soo;Choi Kwan-Yong;Yang Jung-Jin;Lee Sang-Kyou
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.911-920
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    • 2006
  • [ $CD8^+$ ] T Iymphocytes with the cytotoxic activity and capability to release various cytokines are the major players in immune responses against viral infection and cancer. To identify the proteins specific to resting or activated human CD8$^+$ T cells, human CD8$^+$ T cells were activated with anti-CD3+anti-CD28 mAb in the presence of IL-2. The solubilized proteins from resting and activated human CD8$^+$ T cells were separated by high-resolution two-dimensional polyacrylamide gel electrophoresis, and their proteomes were analyzed. Proteomic analysis of resting and activated T cells resulted in identification of 35 proteins with the altered expression. Mass spectrometry coupled with Profound and SWISS-PROT database analysis revealed that these identified proteins are to be functionally associated with cell proliferation, metabolic pathways, antigen presentation, and intracellular signal transduction pathways. We also identified six unknown proteins predicted from genomic DNA sequences specific to resting or activated CD8$^+$ T cells. Protein network studies and functional characterization of these novel proteins may provide new insight into the signaling transduction pathway of CD8$^+$ T cell activation.

Marked Expansion of CD11c+CD8+ T-Cells in Melanoma-bearing Mice Induced by Anti-4-1BB Monoclonal Antibody

  • Ju, Seong-A;Park, Sang-Min;Lee, Sang-Chul;Kwon, Byoung S.;Kim, Byung-Sam
    • Molecules and Cells
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    • v.24 no.1
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    • pp.132-138
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    • 2007
  • 4-1BB (CD137), a member of the tumor necrosis factor receptor superfamily, is expressed on activated T-cells, and 4-1BB signaling due to interaction with 4-1BB ligand or ligation with anti-4-1BB monoclonal antibody (mAb) costimulates T cells. It has been shown that administration of anti-4-1BB mAb induces anti-tumor immunity in mice, but the nature of the cellular subsets responsible for this immunity is uncertain. In this study we found that anti-4-1BB mAb administration to B16F10 melanoma-bearing mice induced marked expansion of $CD11c^+CD8^+$ T-cells in parallel with suppression of pulmonary tumors. The mAb-treated mice produced higher levels of $IFN-{\gamma}$ in their tumor tissues, spleen and lymph nodes than mice exposed to control antibody. When the $CD11c^+CD8^+$ T-cells were purified and re-stimulated in vitro, they produced high levels of the Th1 cytokines, $IFN-{\gamma}$ and IL-2, but low levels of the Th2 cytokines, IL-4 and IL-10. Furthermore, they expressed high levels of 4-1BB and CD107a, a marker of activated cytotoxic T-lymphocytes. Our results suggest that $CD11c^+CD8^+$ T-cells play a role in the anti-tumor immunity induced by anti-4-1BB mAb.

Changes in Phosphate Transporter Activity Evaluated by Phosphonoformic Acid Binding in Cadmium-Exposed Renal Brush-Border Membranes

  • Chung, Jin-Mo;Ahn, Do-Whan;Kim, Kyoung-Ryong;Park, Yang-Saeng
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.5
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    • pp.513-519
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    • 1999
  • Direct exposure of renal tubular brush-border membranes (BBM) to free cadmium (Cd) causes a reduction in phosphate (Pi) transport capacity. Biochemical mechanism of this reduction was investigated in the present study. Renal proximal tubular brush-border membrane vesicles (BBMV) were isolated from rabbit kidney outer cortex by Mg precipitation method. Vesicles were exposed to $50{\sim}200\;{\mu}M\;CdCl_2$ for 30 min, then the phosphate transporter activity was determined. The range of Cd concentration employed in this study was comparable to that of the unbound Cd documented in renal cortical tissues of Cd-exposed animals at the time of onset of renal dysfunction. The rate of sodium-dependent phosphate transport $(Na^+-Pi\;cotransport)$ by BBMV was determined by $^{32}P-Iabeled$ inorganic phosphate uptake, and the number of $Na^+-Pi$ cotransporters in the BBM was assessed by Pi-protectable $^{14}C-labeled$ phosphonoformic acid $([^{14}C]PFA)$ binding. The exposure of BBMV to Cd decreased the $Na^+-Pi$ cotransport activity in proportion to the Cd concentration in the preincubation medium, but it showed no apparent effect on the Pi-protectable PFA binding. These results indicate that an interaction of renal BBM with free Cd induces a reduction in $Na^+-Pi$ cotransport activity without altering the carrier density in the membrane. This, in turn, suggest that the suppression of phosphate transport capacity $(V_{max})$ observed in Cd-treated renal BBM is due to a reduction in $Na^+-Pi$ translocation by existing carriers, possibly by Cd-induced fall in membrane fluidity.

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Anticancer Effect of Activated Natural Killer Cells on Human Non-small Cell Lung Cancer (비소세포성폐암에 대한 자연살해세포의 항암효능)

  • Park, Min-Gyeong;Sung, Hye-Ran;Park, Ji-Sung;Kim, Jee-Youn;Han, Sang-Bae;Lee, Chong-Kil;Yun, Byung-Kui;Song, Suk-Gil
    • YAKHAK HOEJI
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    • v.55 no.3
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    • pp.267-272
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    • 2011
  • Human NK cells, identified 30 years ago based on their ability to spontaneously kill tumor cells, constitute a subset of lymphocytes, which play an important role in the first line of immune defense and the effective function of these cells are enhanced by cytokines. Lung carcinoma has been one of the most commonly diagonosed cancer as well as the leading cause of cancer death in male. Here we provide the evidence that human natural killer cells has inhibitory effects on tumor growth of human lung cancer cell NCI-H460 (non-small cell lung cancer). Enriched NK cell population was obtained by 2 weeks cultivation in interleukin-2(IL-2)-containing medium. The resulting population comprised 26% CD3$^+$ cells, 9% CD3$^+$CD4$^+$ cells, 16% CD3$^+$CD8$^+$ cells, 76% CD56$^+$ cells, 6% CD3$^+$CD56$^+$ cells and 70% CD3$^-$CD56$^+$ cells. Activated NK cells at doese of 2.5, 5, and 10 million cells per mouse inhibited 2%, 12% and 45% of NCI-H460-induced tumor growth in nude mouse xenograft assays, repectively. This result suggests that NK cell-based immunotherapy may be used as an adoptive immunotherapy for lung cancer patients.

Comparison of small bowel findings using capsule endoscopy between Crohn's disease and intestinal tuberculosis in Korea

  • Kim, Yong Gil;Kim, Kyung-Jo;Min, Young-Ki
    • Journal of Yeungnam Medical Science
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    • v.37 no.2
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    • pp.98-105
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    • 2020
  • Background: Little is known about capsule endoscopy (CE) findings in patients with intestinal tuberculosis who exhibit small bowel lesions. The aim of the present study was to distinguish between Crohn's disease (CD) and intestinal tuberculosis based on CE findings. Methods: Findings from 55 patients, who underwent CE using PillCam SB CE (Given Imaging, Yoqneam, Israel) between February 2003 and June 2015, were retrospectively analyzed. Results: CE revealed small bowel lesions in 35 of the 55 patients: 19 with CD and 16 with intestinal tuberculosis. The median age at diagnosis for patients with CD was 26 years and 36 years for those with intestinal tuberculosis. On CE, three parameters, ≥10 ulcers, >3 involved segments and aphthous ulcers, were more common in patients with CD than in those intestinal tuberculosis. Cobblestoning was observed in five patients with CD and in none with intestinal tuberculosis. The authors hypothesized that a diagnosis of small bowel CD could be made when the number of parameters in CD patients was higher than that for intestinal tuberculosis. The authors calculated that the diagnosis of either CD or intestinal tuberculosis would have been made in 34 of the 35 patients (97%). Conclusion: The number of ulcers and involved segments, and the presence of aphthous ulcers, were significantly higher and more common, respectively, in patients with CD than in those with intestinal tuberculosis. Cobblestoning in the small bowel may highly favor a diagnosis of CD on CE.

Preparation and Bioevaluation of 177Lu-labelled Anti-CD44 for Radioimmunotherapy of Colon Cancer

  • Lee, SoYoung;Hong, YoungDon;Jung, SungHee;Choi, SunJu
    • Journal of Radiation Industry
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    • v.9 no.4
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    • pp.187-192
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    • 2015
  • CD44 is a particular adhesion molecule and facilitates both cell-cell and cell-matrix interactions. In particular, splice variants of CD44 are particularly overexpressed in a large number of malignancies and carcinomas. In this study, the $^{177}Lu$-labelled CD44 targeting antibody was prepared and bioevaluated in vitro and in vivo. Anti-CD44 was immunoconjugated with the equivalent molar ratio of cysteine-based DTPA-NCS and radioimmunoconjugated with $^{177}Lu$ at room temperature within 15 minutes. The stability was tested in human serum. An in vitro study was carried out in HT-29 human colon cancer cell lines. For the biodistribution study $^{177}Lu$-labelled anti-CD44 was injected in xenograft mice. Anti-CD44 was immunoconjugated with cysteine-based DTPA-NCS and purified by a centricon filter system having a molecular cut-off of 50 kDa. Radioimmunoconjugation with $^{177}Lu$ was reacted for 15 min at room temperature. The radiolabeling yield was >99%, and it was stable in human serum without any fragmentation or degradation. The radioimmunoconjugate showed a high binding affinity on HT-29 colon cancer cell surfaces. In a biodistribution study, the tumor-to-blood ratio of the radioimmunoconjugate was 43 : 1 at 1 day post injection (p.i) in human colon cancer bearing mice. The anti-CD44 monoclonal antibody for the targeting of colon cancer was effectively radioimmunoconjugated with $^{177}Lu$. The in vitro high immunoactivity of this radioimmunoconjugate was determined by a cell binding assay. In addition, the antibody's tumor targeting ability was demonstrated with very high uptake in tumors. This radioimmunoconjugate is applicable to therapy in human colon cancer with highly expressed CD44.

β-carotene regulates cancer stemness in colon cancer in vivo and in vitro

  • Lee, Kyung Eun;Kwon, Minseo;Kim, Yoo Sun;Kim, Yerin;Chung, Min Gi;Heo, Seung Chul;Kim, Yuri
    • Nutrition Research and Practice
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    • v.16 no.2
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    • pp.161-172
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    • 2022
  • BACKGROUND/OBJECTIVES: Colorectal cancer (CRC) is the third most common cancer worldwide and has a high recurrence rate, which is associated with cancer stem cells (CSCs). β-carotene (BC) possesses antioxidant activity and several anticancer mechanisms. However, no investigation has examined its effect on colon cancer stemness. MATERIALS/METHODS: CD133+CD44+ HCT116 and CD133+CD44+ HT-29 cells were isolated and analyzed their self-renewal capacity by clonogenic and sphere formation assays. Expressions of several CSCs markers and Wnt/β-catenin signaling were examined. In addition, CD133+CD44+ HCT116 cells were subcutaneously injected in xenograft mice and analyzed the effect of BC on tumor formation, tumor volume, and CSCs markers in tumors. RESULTS: BC inhibited self-renewal capacity and CSC markers, including CD44, CD133, ALDH1A1, NOTCH1, Sox2, and β-catenin in vitro. The effects of BC on CSC markers were confirmed in primary cells isolated from human CRC tumors. BC supplementation decreased the number and size of tumors and delayed the tumor-onset time in xenograft mice injected with CD133+CD44+ HCT116 cells. The inhibitory effect of BC on CSC markers and the Wnt/β-catenin signaling pathway in tumors was confirmed in vivo as well. CONCLUSIONS: These results suggest that BC may be a potential therapeutic agent for colon cancer by targeting colon CSCs.

The Effects of Sinapis Semen, Raphani Semen, and mixture decoction on the Asthmatic Mouse Model (백개자, 나복자 및 두 배합 약물의 천식 동물 모델에 대한 효과)

  • Kim, Chang-Min;Lee, Young Cheol;Lee, Jang-Cheon
    • The Korea Journal of Herbology
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    • v.28 no.6
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    • pp.15-23
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    • 2013
  • Objectives : To clarify the possible effects of Sinapis Semen and Raphani Semen on the development of pulmonary eosinophilic inflammation in a asthmatic mouse model. Methods : BALBav/c mice were sensitized to OVA followed intratracheally and by aerosol allergene challenges. We investigated the effect of Sinapis Semen and Raphani Semen on airway hyperresponsiveness, eosinophiic infitratio, immune cell phenotype, The2 cytokine product, and OVA-spedific IgE production. Results : Total lung cells, eosinophils, and lung leukocytes, OVA specific IgE levels, and Th 2cytokine levels such as IL-5, IL-13, IL-17, TNF-alpha, and eotaxin in BALF were reduced compared with those of OVA sensitized asthma mice (control). The absolute numbers of $CD3^+$, $CD3^+/CD69^+$, $CD3^-/CCR3^+$, $CD4^+$, $CD8^+$, $Gr-1^+/CD11b^+$, $B220^+/CD22^+$, $B220^+/IgE^+$ cells in lung tissiues significantly reduced compared to those of control. Specially total lung cells in BALF and the absolute number of $CD3^+/CD69^+$ and, $B220^+/IgE^+$ cells in lung tissiue effectively reduced in Sinapis Semen plus Raphani Semen compared to those of Sinapis Semen and Raphani Semen. Conclusions : These results indicate that Sinapis Semen plus Raphani Semen has deep inhibitory effects on airway inflammation and hyperresponsiveness in asmatic mouse model and also has effect of suppression of IL-5, IL-13, IL-17, OVA specific IgE production in BALF. The results verified that Sinapis Semen, Raphani Semen, and Sinapis Semen plus Raphani Semen could act as a immunomodulator which possess anti-inflammatory and anti-asthmatic property by modulating the relationship of Th1/Th2 cytokine imbalance.

Changes of enzyme activity in the hemolymph and hepatopancreas of the abalone, Haliotis discus hannai (Ino, 1953) exposed to cadmium (카드뮴 노출에 따른 북방전복, Haliotis discus hannai (Ino, 1953) 의 hemolymph 및 hepatopancreas의 효소활성의 변화)

  • Min, Eun-Young;Lee, Jung Sick;Kwak, Ihn-Sil;Kim, Jae Won;Kang, Ju-Chan
    • The Korean Journal of Malacology
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    • v.30 no.1
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    • pp.41-49
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    • 2014
  • This study was conducted to investigate the effects of cadmium (Cd) exposure on biochemical factors in the hemolymph and hepatopancreas of the abalone, Haliotis discus hannai. The abalone were exposed to 0, 5, 10, 20 and 40 ${\mu}g/L$ Cd for 4 weeks. The phenoloxidase (PO) activity was decreased in hemolymph of abalone exposed to 40 Cd ${\mu}g/L$ for 4 weeks compared to the control (P < 0.05). The hemolymph enzymes, alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were markedly elevated in 40 Cd ${\mu}g/L$ after 4 weeks. The hemolymph calcium concentrations were significantly decreased in 20 and 40 Cd ${\mu}g/L$ for 4 weeks. Hepatopancreas superoxide dismutase (SOD) and catalase (CAT) activities were significantly increased by Cd. SOD was increased in both 20 and 40 Cd ${\mu}g/L$ and CAT, in 40 Cd ${\mu}g/L$ after 2 weeks (P < 0.05). These results suggested that the abalone SOD and CAT including PO may serve as a protective mechanism against oxidative stress by Cd. We conclude that a Cd concentration, 40 ${\mu}g/L$ in water may curtail hemolymph homeostasis and anti-oxidative reactions in abalone hepatopancreas. From these results, these biochemical factors may represent a convenient method of monitoring heavy metal pollution in coastal areas.

Dietary intake and body burden of cadmium and lead among Korean college students (일부 대학생들의 카드뮴과 납 섭취량 및 체내부하)

  • Moon, Chan-Seok;Paik, Jong Min
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.21 no.1
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    • pp.25-32
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    • 2011
  • Objectives: This study aims to evaluate the dietary intake and blood concentrations for examination of exposure levels and exposure characteristics of Cd and Pb via food among college students aged 20 years living in Busan area. Methods: Food duplicate per a day and venous blood samples were accepted from healthy man and women college students. The samples were analyzed with graphite furnace atomic absorption spectrophotometer. Results: Dietary intake of Cd and Pb in the participants were $17.5{\mu}g\;Cd/day$, $21.7{\mu}g\;Pb/day$. Blood concentrations were $2.01{\mu}g\;Cd/L$, $4.32{\mu}g\;Pb/dL$, respectively. Conclusions: Smoking and drinking habits might be high influential factors in 20 years college students. Dietary intake of Cd and Pb, however, from food group of seaweed, or fish and shellfish in 20 years college students were lower than that in any other age group.