• IMMUNE NETWORK
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• v.10 no.6
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• pp.239-246
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• 2010

Background: Monoclonal antibodies (mAbs) recognizing Class III epitope of CD34 are essential for flow cytometric diagnosis of leukemia. Methods: 27H2 mAb was developed from a mouse alternatively immunized with human acute leukemia cell lines, KG1 and Molm-1. Using flow cytometric analysis of various leukemic cell lines and peripheral blood, immunohistochemical study of frozen tonsil, we characterized 27H2 mAb. Antigen immunoprecipitated with 27H2 mAb immunobloted with anti-CD34 mAb. A case of bone marrow sample of acute lymphoblastic leukemia (ALL) patient was obtained at CBNU Hospital. For epitope identification enzyme treatment with neuraminidase and O-sialoglycoprotein endopeptidase (OSGE) and blocking assay with known classIII mAb (HPCA-2) were done. Results: Only KG1 and Molm-1 revealed positive immunoreactivity. Immunohistochemical staining disclosed strong membranous immunoreactivity on high endothelial venules. Antigen immunoprecipitated by 27H2 mAb showed approximately 100 kDa sized band immunoblotted with anti-CD34 under non-reducing conditions. Epitope recognized by 27H2 mAb disclosed resistancy to both neuraminidase and OSGE treatment and completely blocked with known class III mAb preincubation. CD34 positive leukemic cells in BM of pre B cell ALL patient detected by FITC-conjugated 27H2 and HPCA-2 were identified with similar sensitivity. Conclusion: A novel murine mAb recognizing class III epitope of human CD34 with high affinity, which is useful for flow cytometric diagnosis of leukemia, was developed.

• Journal of Industrial Technology
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• v.25 no.B
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• pp.241-251
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• 2005

Exopolysaccharide (CBP) from submerged culture broth of Ganoderma lucidum mycelium and the water soluble (BWS) and water insoluble (BWI) fractions of CBP were prepared by gel filtration. Antitumor activity and effects on proliferation and differenciation of human cancer cells and mouse NIH 3T3 cells were studied. Cytotoxicity test of CBP, BWS and BWI fractions on human cancer cell lines was performed by using sulforhodamine B (SRB) assay. A549 (lung carcinoma), Colo320 DM and HSR (colon carcinoma), and NIH 3T3 cells were used. BWI fraction showed the strongest cytotoxicity (maximum 20% survival) to all human cells tested. However it did not induced apoptosis. Interestingly BWI fraction did not exert cytotoxic effect on NIH 3T3 cells at low concentration of cells ($5{\times}10^4$) but strong toxic effect at high concentration of cells($5{\times}10^5$) which showed transformed morphology. These results suggest that BWI may have cancer cell specific anticancer activity. However, BWI fraction did not effect the amount of pRb and c-myc protein, which implied that BWI fraction did not act at the early stage of signal transduction pathway. CBP fraction induced differenciation of human leukemic cell line, HL-60 cells suggesting the carcinogenesis prevention of normal cell and possible induction of normalization for cancer cell.

• IMMUNE NETWORK
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• v.6 no.4
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• pp.179-184
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• 2006

Background: Identification of antigen-specific T cells has yielded valuable information on pathologic process and the disease state. Assays for quantification of inflammatory cytokines or lytic-granule molecules have been generally used to evaluate antigen specific T cell response, however their applicability have been hampered due to the limited source of autologous antigen-presenting target cells (APC). Methods: K562, a leukemic cell line deficient of human leukocyte antigen (HLA), was transfected with a gene encoding HLA-A*02 (K562/ A*02) and its function as stimulator cells in inducing activation of HLA-matched T cells was evaluated by IFN-${\gamma}$ enzyme linked immunospot (ELISPOT) assay. Results: The stable transfectant K562/ A*02 pulsed with HLA- A*02 restricted peptide could specifically induce IFN-${\gamma}$ secretion by CD8+ T cells compared to no detectable secretion by CD4+ T cells. However, CD56+ NK cells secreted IFN-${\gamma}$ in both K562/ A*02 with peptide and without peptide. The number of IFN-${\gamma}$ secreted CD8+ T cells was increased according to the ratio of T cells to K562 and peptide concentration. Formalin-fixed K562/ A*02 showed similar antigen presenting function to live K562/ A*02. Moreover, K562/ A*02 could present antigenicpeptide to not only A*0201 restricted CD8+ T cells but also CD8+ T cells from A*0206 donor. Conclusion: These results suggest that K562/ A*02 could be generally used as target having specificity and negligible background for measuring CD8+ T cell responses and selective use of K562 with responsder matched HLA molecules on its surface as APC may circumvent the limitation of providing HLA-matched autologous target cells.

• Molecules and Cells
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• v.46 no.10
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• pp.611-626
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• 2023

Acute myeloid leukemia (AML) is a heterogeneous disease caused by distinctive mutations in individual patients; therefore, each patient may display different cell-type compositions. Although most patients with AML achieve complete remission (CR) through intensive chemotherapy, the likelihood of relapse remains high. Several studies have attempted to characterize the genetic and cellular heterogeneity of AML; however, our understanding of the cellular heterogeneity of AML remains limited. In this study, we performed single-cell RNA sequencing (scRNAseq) of bone marrow-derived mononuclear cells obtained from same patients at different AML stages (diagnosis, CR, and relapse). We found that hematopoietic stem cells (HSCs) at diagnosis were abnormal compared to normal HSCs. By improving the detection of the DNMT3A R882 mutation with targeted scRNAseq, we identified that DNMT3A-mutant cells that mainly remained were granulocyte-monocyte progenitors (GMPs) or lymphoid-primed multipotential progenitors (LMPPs) from CR to relapse and that DNMT3A-mutant cells have gene signatures related to AML and leukemic cells. Copy number variation analysis at the single-cell level indicated that the cell type that possesses DNMT3A mutations is an important factor in AML relapse and that GMP and LMPP cells can affect relapse in patients with AML. This study advances our understanding of the role of DNMT3A in AML relapse and our approach can be applied to predict treatment outcomes.

• YAKHAK HOEJI
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• v.40 no.2
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• pp.117-125
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• 1996

3'-Deamino-3-hydroxy-4'-fluoro- or 3'-deamino-3'-hydroxy-4'-azido-daunorubicin(6,8) and -doxorubicin(7,9) have been synthesized, respectively. Compounds 7,8 and 9 were mo re cytotoxic than daunorubicin(1) and doxorubicin(2) against L1210 murine leukemic cell in vitro. When administered intraperitoneally for 9 days starting 1 day after tumor inoculation, compounds 7(T/C 605%) and 9(T/C 488%) showed significant antitumor activity for ip-inoculuated L1210 murine leukemia at wide range of doses.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.336.3-337
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• 2002

We previously reported that a synthetic naphthoquinone analog. 2.3-dichloro-5.8-dihydroxy-1, 4-naphthoquinone (NA). effectively induces apoptosis in human leukemic HL-60 cells. However. the cellular mechanism by which NA induces cell death remain unclear. In this study. we show that NA induces activation of capases. release of cytochrome c and upregulation of proapoptotic Bax protein. Futhermore. NA suppressed phosphorylation of Akt and Bad. suggesting that Akt regulates NA-induced apoptosis. Expresson of a dominant negative Akt enhancde NA-induced apoptosis. suggesting that naphthoquinone analog induces apoptosis through activating proapoptotic pathway and by the inactivation of antiapoptotic pathway.

• Korean Journal of Clinical Pharmacy
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• v.19 no.2
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• pp.89-95
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• 2009

Gemtuzumab ozogamicin (GO) is an antibody-targeted chemotherapeutic agent consisting of calicheamicin, a potent cytotoxic antibiotic linked to a recombinant humanized anti CD33 monoclonal antibody directed against the CD33 antigen present on leukemic myeloblasts in most patients with acute myeloid leukemia (AML). GO is indicated for the treatment of patients with CD33 positive AML in first relapse who are 60 years of age or older and who are not considered candidates for cytotoxic chemotherapy. GO has shown moderate activity as a single agent in patients with CD33-positive refractory or relapsed acute myeloid leukaemia, with more promising results in acute promyelocytic leukaemia. The side effect profile may be an improvement on conventional chemotherapy, except for a higher frequency of veno-occlusive disease or sinusoidal obstructive syndrome, especially after a subsequent haematopoietic stem cell transplantation. Because of the different mechanisms of action and non-overlapping toxicities, the integration of this immunoconjugate with standard chemotherapy is a rational approach.

• Journal of Veterinary Clinics
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• v.3 no.1
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• pp.265-276
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• 1986

Pathological study was made of 173 gallbladders among 652 slaughter pigs. The size and the pH of the bile juice were made of 50 among 173 gallbladders with lesions and 50 normal gallbladders as control. The results are summerized as follows : 1. The histological lesions were shown hyperemia and congestion(83.2%), Iymphocytic infiltration(37.5%), Iympoid hyperplasia(27%), epithelial downgrowth(27%), erosion and ulcer(22%), papillary hyperplasia (20.2%), pericholecystic edema(19.5%), hemorrhage(11%), and leukemic cell infiltration(0.6%). 2. In the gallbladders with lesions mean size was 4.26${\times}$7.88cm in diameter and mean pH is 7.2. In the control means size and mean pH were 4.57${\times}$8.13cm in diameter and 7.1. The difference of size between gallbladders with lesions and these as control was not statistically significant. The pH of bile juice between two showed also no statistically significant difference.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.158.2-158.2
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• 2003

Vitamin K-related analogs induce growth inhibition in various cancer cell lines. We report that 2,3-dichloro-5,8-dihydroxy-1, 4-naphthoquinone (DDN), a naphthoquinone analog, induces mitochondria-dependent apoptosis in human promyeloid leukemic HL-60 cells. DDN induced cytochrome c release, cleavage of Bid, and activation of caspases -8, -9 and -3. Cleavage of Bid, the caspase-8 substrate, was inhibited by the broad caspase inhibitor zVAD-fmk, whereas cytochrome c release was not affected by zVAD-fmk. (omitted)

• Biomolecules & Therapeutics
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• v.18 no.2
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• pp.178-183
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• 2010

Costunolide is an active compound isolated from the stem bark of Magnolia sieboldii, and is considered a potential therapeutic for the treatment of various cancers. In this study, we investigated the underlying mechanism whereby costunolide induces the apoptosis of human leukemia cells. Using apoptosis analysis and quantitative reverse transcription-polymerase chain reaction (RT-PCR) results obtained during this study show that costunolide is a potent inducer of apoptosis and that it is triggered due to the premature activation of Cdc2. $G_1$-synchronized cells, which cannot undergo mitosis, were found to be more sensitive to costunolide, and Cdc2 mRNA levels were increased by costunolide treatment. Furthermore, the Cdk inhibitors, olomucine and butyrolactone I, were found to suppress costunolide-induced apoptosis. In addition, the PKC activator TPA rescued cells from cell death by costunolide, and this was prevented by the PKC inhibitor staurosporin. The present study suggests that costunolide induces the apoptosis of HL-60 leukemic cells by modulating cyclin-dependent kinase Cdc2.