• Title/Summary/Keyword: myeloma cell

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Monoclonal Antibody-Based Indirect-ELISA for Early Detection, Diagnosis and Monitoring of Epiphytic Didymella bryoniae in Cucurbits.

  • Lee, Seon-Chul;Shim, Chang-Ki;Kim, Dong-Kil;Bae, Dong-Won;Kyo, Seo-Il;Kim, Hee-Kyu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.133.1-133
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    • 2003
  • Gummy stem blight, caused by Didymella bryoniae occurs exclusively on cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. Through this study, we optimized cultural conditions for mass-production of pycnidiospore by Metal Halide Lamp irradiation. In brief, the mycelial was cultured at $26^{\circ}C$ on PDA, for 2 days under the darkness, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$, a great number of pycnidia was simultaneously formed. Thus produced pycnidiospores were used as immunogen. From fusions of myeloma cell (v-653) with splenocytes from immunifed mice were car ried out. And, two hybridoma cell lines that recognized the immunogen Didymella bryoniae were obtained. One Monoclonal Antibody, Db1, recognized the supernatant and the other monoclonal antibody, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid two MAb Db1 and Db15, were immunotyped and identified as IgG1 and IgG2b, respectively. Titer of MAb Db1 and MAb Db15 was measured absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with Didymella bryoniae but none reacted with other of fungi and CMV, CGMMV Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{3}$ well by indirect ELISA characterization of the MAb Db1, Db15 antigen by heat and protease treatments show that the epitope recognized by the MAb Bb1, Db15 were a glycoprotein.

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Extramedullary plasmacytoma in the maxillary sinus: a case report (상악동에 발생한 골수외 형질세포종: 증례보고)

  • Kim, Hyoung-Keun;Hahm, Tae-Hoon;Nam, Woong;Cha, In-Ho;Kim, Hyung-Jun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.6
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    • pp.543-547
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    • 2010
  • Plasmacytoma is a rare malignant neoplasm in the head and neck region and comprises approximately 3% of all plasma cell tumors. This lesion is a unifocal, monoclonal, neoplastic proliferation of plasma cells that usually arises within the bone. Infrequently, it is observed in soft tissue, in which case, the term extramedullary plasmacytoma is used. Approximately 80-90% of extramedullary plasmacytomas involve the mucos-Associated-Lymphoid Tissue of the upper airways with 75% of these involving the nasal and paranasal regions. The plasmacytoma is usually detected in adult males, with an average age at diagnosis of 55 years. The male-to-female ratio is 3:1.Radiographically, the lesion may be seen as a well-defined, unilocularradioluceny with no evidence of a sclerotic border. Some investigators believe that this lesion represents the least aggressive part of the spectrum of plasma cell neoplasms that extend to multiple myeloma.Therefore, plasma cytoma is believed to have clinical importance. We report a case of extramedullary plasmacytoma in the right maxillary sinus of a 59-year-old male with review of the relevant literature.

Characterization of Monoclonal Antibodies against Heavy and Light Chains of Flounder (Paralichthys olivaceus) Immunoglobulin

  • Jang, Han-Na;Woo, Jong-Kyu;Cho, Young-Hye;Kyong, Seo-Bong;Choi, Sang-Hoon
    • BMB Reports
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    • v.37 no.3
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    • pp.314-319
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    • 2004
  • Flounder (Paralichthys olivaceus) Immunoglobulins (Igs) were purified from the serum of mouse IgG-immunized flounder by using affinity chromatography. Under denaturing conditions in SDS-PAGE, the flounder Igs appeared to be composed of 2 heavy (H) chains (72 and 77 kDa) and two light (L) chains (26 and 28 kDa). Monoclonal antibodies (MAbs) were produced by the fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells that were previously sensitized against affinity-purified flounder Igs. In a Western blot analysis, the produced MAbs, FIM511, FIM519, and FIM562 recognized both the 72 and 77 kDa H chains, 26 kDa, and 28 kDa L chain, respectively. Mouse antiserum against flounder Igs reacted more strongly with the L chain of 28 kDa than with 26 kDa, suggesting that the 28 kDa molecule is more immunogenic than the 26 kDa L chain molecule. In a FACS analysis, the ratios of the Ig+ cell population in the flounder head kidney and spleen cells were 49% and 24%, respectively. Unexpectedly, however, the ratios of the Ig+ B-like cell population in the flounder were not significantly augmented, even after the immunization of an immunogenic antigen. This suggests that the humoral immune response in fish could be considerably different from that in mammals. The produced MAbs in this study would be useful in characterizing flounder Ig+ B-like cells and in developing flounder Ig detecting an immunoassay system.

Production of Monoclonal Antibody to Infectious Laryngo- tracheitis Virus by Cell Fusion (닭 전염성 후두기관염 바이러스에 대한 단크론성 항체생산)

  • Chung Ok Choi;Chung Gil Lee;Sung Man Cho;Soo Hwan An;Joon Hun Kwon
    • Korean Journal of Poultry Science
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    • v.15 no.3
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    • pp.199-206
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    • 1988
  • A total of 3 hybridoma clones producing monoclonal antibody (MCA) against infectious laryngotracheitis virus (ILTV) was established by somatic cell hybridization between mouse myeloma cells and spleen, cells from mice immunized with ILTV. The MCAs were screened by the indirect flourescent antibody (IFA) staining and the specific hybridomas were cloned by limiting dilution method. The MCAs produced by the 3 hybriomas were all classified as immunogloblin G and found to be reacting against common antigen(s) of high and low pathogenic ILTV examined. The titer of these antibodies in mouse ascitic fluid was from $10^5$ to $10^6$. Indirect fluorescent antibody test using these antibodies was found to be quite effective for the detection of ILTV from infected chickens being the most sensitive among the test methods adopted.

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Solitary Plasmacytoma of the Skull - A Case Report - (두개골에 발생한 고립성 형질세포종 - 증례보고 -)

  • Han, Jeong Hoon;Park, Hae Kwan;Min, Chang Ki;Cho, Jung Ki;Park, Sung Chan;Cho, Kyung Keun;Lee, Kyung Jin;Rha, Hyoung Kyun;Choi, Chang Rak;Kang, Joon Ki
    • Journal of Korean Neurosurgical Society
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    • v.29 no.5
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    • pp.701-705
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    • 2000
  • Solitary plasmacytomas are rare and account for 5-10% of all plasma cell disorders. These tumors are categorized as solitary plasmacytomas of bone(osseous) or extramedullary plasmacytomas(non-osseous). About a half of solitary plasmacytomas of bone occur in the spine but rarely in the skull. We report a case of solitary plasmacytoma of the skull presented with a painless palpable left parietal calvarial mass in an otherwise asymptomatic 38- year-old man. Skull radiographs showed a large radiolucent lesion with well defined non-sclerotic margins. Computed tomograph scan demonstrated a markedly enhancing mass extending from the epidural to the subcutaneous space. The patient underwent surgery and tumor was completely excised. Pathological examination showed tumor to be a plasmacytoma synthesizing IgG. Postoperatively, the patient received radiotherapy. There was no evidence of systemic involvement on postoperative laboratory wokups. Our recommended treatment is a complete surgical excision combined with postoperative radiation therapy. The patient should be follwed carefully for more than 10 years because of either local recurrence or possible progression to multiple myeloma.

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Monoclonal Antibody-Based Indirect-ELISA for Early Detection and Diagnosis of Epiphytic Didymella bryoniae in Cucurbits

  • Lee, Sun-Cheol;Han, Ki-Soo;Lee, Jung-Han;Kim, Dong-Kil;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • v.19 no.5
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    • pp.260-265
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    • 2003
  • Gummy stem blight caused by Didymella bryoniae occurs exclusively in cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. In this study, cultural conditions for the mass-production of pycnidiospore by Metal Halide (MH) lamp irradiation were maximized. The mycelia were cultured at $26^{\circ}C$ on PDA for 2 days under dark condition, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$. Results show that a great number of pycnidia were simultaneously formed. The pycnidiospores produced were then used as immunogen. Fusions of myeloma cell (v-653) with splenocytes from immunized mice were carried out. Two hybridoma cell lines that recognized the immunogen D. bryoniae were obtained. One monoclonal antibody (MAb), Dbl, recognized the supernatant while another MAb, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid of the two MAb, Dbl and Db15, the immunotypes of which were identified as IgG1 and IgG2b, respectively. Titers of MAb Dbl and MAb Db15 were measured and the absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with D. bryoniae but none reacted with other viral isolates, Cucumber mosaic virus and Cucumber green mottle mosaic virus. Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{-3}$/well by indirect ELISA. Characterization of the MAbs Dbl, Db15 antigen by heat and protease treatments, which suggests that the epitope recognized by these two MAbs was glycoprotein.

Preparation and Characterization of Cell Hybrids Producing a Monoclonal Antibody to Human Fibroblast Interferon (Hu IFN-$\beta$) (사람 선유아세포 인터페론(Hu IFN-$\beta$)에 대한 단 Clone성 항체생산세포의 조작과 그 성질에 관한 연구)

  • 김현수;현형환;최경희;문홍모;유무영
    • Microbiology and Biotechnology Letters
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    • v.14 no.3
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    • pp.219-223
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    • 1986
  • In order to preparr the hybridoma cells which produce a monoclonal antibody to human fibroblast interferon(Hu IFN-$\beta$), spleen cells from BALB/cmice immunized with the purified Hu IFN-$\beta$ were fused with NS-O cells, a myeloma cell line. Forty hybrids with high titer among 1300 hybrids Isolated by an ELISA screening method were subcloned using the soft agarose cloning and limiting dilution methods, and 11 hybrids were selected. As a result of iso-typing the hybrids using the mouse monoclonal typing kit, two hybridomas were found to produce 1gG 2a type of monoclonal an-tibodies. The ascites fluid from nude mice inoculated intraperitoneally with the above hybridomas was removed and purified using a protein A-Sepharose CL-4B. Monoclonal antibody was proven to have only the heavy and light chains on SDS-polyacrylamide gel electrophoresis.

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A Case of Extramedullary Plasmacytoma in Palatine Tonsil Achieved Complete Remission by Radiotherapy (국소 방사선 치료에 완전 관해된 구개편도에 발생한 골수외 형질세포종)

  • Lee Jung-Hwa;Kim Ki-Seong;Kim Hyo-Jung;Park Hee-Chul;Bae Hoon-Sik;Ha Jun-Wook;Zang Dae-Young
    • Korean Journal of Head & Neck Oncology
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    • v.21 no.2
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    • pp.165-169
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    • 2005
  • Extramedullary plasmacytoma(EMP) is a rare tumor that originates as a clone of malignant transformed plasma cells. EMPs make up 4% of all plasma cell tumors and 90% of EMP cases occur in the head and neck area. The diagnosis of EMP is based on the morphologic and immunophenotypical finding of a localized collection of monoclonal plasma cells in the absence of plasma cell proliferation elsewhere. Patients with EMP can be treated by surgery or radiotherapy alone or in combination. EMP has good prognosis but long-term follow-up is critical because local recurrence or systemic progression to multiple myeloma may develop more than 30 years after the initial diagnosis of EMP. We report a case of EMP in palatine tonsil of 21-year old man who was treated successfully by radiotherapy alone.

Expression Analysis of the Ligand to Ly-6E.1 Mouse Hematopoietic Stem Cell Antigen

  • Hwang, Dae-Youn;Min, Dul-Lei;Sonn, Chung-Hee;Chang, Mi-Ra;Lee, Mi-Hyun;Paik, Sang-Gi;Kim, Young-Sang
    • Animal cells and systems
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    • v.1 no.1
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    • pp.157-164
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    • 1997
  • Ly-6E.1 antigen was proposed as a regulatory molecule of T lymphocyte activation, a hematopoietic stem cell marker, a memory cell marker, and an adhesion molecule. Though there were several reports suggesting the presence of Ly-6 ligand, the characterization of the ligand was not yet performed, As an attempt to screen the expression of Ly-6E.1 ligand, we prepared a probe for detecting Ly-6E.1 ligand by producing a fusion protein between Ly-6E.1 and $hlgC_{r1}$, A mammalian cell expression vector with Ly-6E.$1/hlgC_{r1}$ chimeric cDNA was transfected in SP2/0-Ag14 myeloma cells, and stable transfectants were selected. The fusion protein was produced as a dimer and maintained the epitopes for monoclonal antibodies specific for Ly-6E.1 and for anti-human lgG antibody. The purified fusion protein through Gammabind G column was used for FACS analyses for the expression of Ly-6E.1 ligand. The fusion protein interacted with several cell lines originating from B cells, T cells, or monocytes. The fusion Protein also strongly stained bone marrow, lymph node, and spleen cells, but thymic cells weakly, if any. The staining was more obvious in C57BL/6 $(Ly-6^b)$ than Balb/c $(Ly-6^a)$ mice. These results suggest that the interaction of Ly-6E.1 with Ly-6E.1 ligand may function both in the stem cell environment and in the activation of mature lymphocytes. The fusion protein may be a valuable tool in characterization of biochemical properties of the Ly-6E.1 ligand and, further, in isolating its cDNA.

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Anticancer Effect of Eriobotrya japonica Lindl by Specificity Test with Several Cancer Cell Lines (비파(枇杷)의 항암효과에 대한 암세포특이성 검정)

  • Whang, Tay-Eak;Lim, Hyun-Ock;Lee, Jae-Wa
    • Korean Journal of Medicinal Crop Science
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    • v.4 no.4
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    • pp.314-320
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    • 1996
  • Antitumouric effect of loquat was investigated treating the extract from leaves, stems, fruits or seeds to normal and cancer cell lines to check by MTT method wheather the cancer cells are spec­ifially attacked. The results are summarized as follows. Water of MeOH extract from each organ were applied to human normal and cancer cell lines, SNU-1 and SNU-C4. The water extract from fruit flesh gave no dffect to normal cell lines by killed all the cancer cell lines. The water extract from fruit flesh was purified by Sephadex LH-20 and separated into 9 fractions which were than applied to 8 cancer cell lines. The eighth fraction out of the 9 fractions gave no effect to normal cells but exerted specific cytotoxicity to breast cancer, stomach and liver cancer cells. The eighth fraction was orally administed and injected to 10 mice each suffering from the abdominal cancer induced by myeloma cells, SP2/0-Ag14. In the groups received the treatment, only one mouse each died in 2 months but the rest survived until the end of the experimental period, which those in the control plot died in 10 to 13 days. The present results confirmed that loquat contained some substance that had specific cytotoxicity to human cancer cells.

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