• Title/Summary/Keyword: myeloma cells

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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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Analysis of Plant Hormones by Immunoassay I . Production of Monoclonal Antibodies to Indole- 3-acetic acid (면역측정법을 이용한 식물 홀몬의 분석 I. 옥신(IAA)에 대한 단크론 항체 생산)

  • 황태익;임현옥;이재와
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.36 no.6
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    • pp.506-512
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    • 1991
  • Monoclonal antibodies (mAb) to indole-3-acetic acid (IAA) were produced and characterized. Spleen cells from mouse immunized with IAA coupled to bovine serum albumin were fused with SP2/0-Ag14 myeloma cells. Three clones secreted specific antibodies to IAA were established to hybridoma cell lines and designated WLI-G1, WLI-G3 and WLI-Ell. The antibodies produced were classified into IgG, types and revealed the high degree of specificity by cross-reaction in the IAA derivatives and its analogues. In the IAA-ELISA with mAb, the measuring range of the assay was 1-500 p mol, and Ka and binding capacity calculated from Scatchard plot were 6.7 X 10$^{-10}$ L/M and 6 x 10$^{-10}$ L/M respectively. The ELISA with mAb can be used to quantitate IAA directly in crude plant eatract. The results showed that the immunoassay was easy and sensitive method to perform and applicate for quantitative analysis of IAA in plant.

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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.

Studies on antigenicity and production of monoclonal antibody for diagnosis of canine heartworm(Dirofilaria immitis) (개 심장사상충(Dirofilaria immitis) 진단을 위한 항원성 조사 및 단크론항체 생산)

  • Lee, Cheol-soon;Jee, Cha-ho
    • Korean Journal of Veterinary Research
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    • v.40 no.1
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    • pp.130-137
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    • 2000
  • In order to diagnose canine heartworm infection by antigen capture ELISA, the crude somatic(S), partial somatic(below 45kDa) and excretory/secretory(E/S) antigen of adult heartworm were identified and the antigenicity was examined by silver stain, immunoblot and ELISA. Then, production of monoclonal antibody to specific antigen carried out in this experiment. The bands to S antigen and E/S antigen were recognized between 10 and 200kDa and common bands were recognized strongly 14, 18, 28, 43kDa by silver stain. By western blot analysis, fractions to S antigen were recognized 14, 16, 18, 20, 24, 28, 32, 43, 50, 55kDa, etc. and only a 14kDa to E/S antigen in positive sera which were positive in modified Knott's test and necropsy. In ELISA, the positive sera reacted to antigens(SA, $SA_{45}$, E/S) were significantly different from negative sera by Student's t-test(p<0.05). Four hybridoma cell lines(14, 16, 17, 32kDa) than produce specific monoclonal antibodies for these antigens were obtained by immunizing BALB/c mice with a partially purified somatic antigen (below 45kDa) preparation, by fusing spleen cells with SP2/O cell myeloma cells, and by screening cell culture supernatants for antibody. In these results, it was confirmed that partial somatic antigen(below 45kDa) or E/S antigen can be used for serologic diagnosis of heartworm infection and monoclonal antibody reacting with specific antigen(14kDa) can be used for antigen capture ELISA in prepatent period of canine heartworm infection.

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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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A Clinical Study on Multiple Myeloma (다발성 골수종의 임상적 고찰)

  • Hwang, Hyeong-Ki;Lee, Choong-Ki;Hyun, Myung-Soo;Shim, Bong-Sup;Lee, Hyun-Woo
    • Journal of Yeungnam Medical Science
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    • v.8 no.2
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    • pp.106-113
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    • 1991
  • A clinical review of 31 cases of multiple myeloma which were diagnosed by criteria of the SWOG between May 1983 and February 1990 at Yeungnam University Hospital was done. The results were as followings : 1. The peak incidence was in 7th decade and male to female ratio was 1.8 : 1. 2. The most common presenting symptom at first diagnosis was bone pain (58%), but fever, dyspnea, dizziness and palpable mass were also noted. 3. The distribution of laboratory findings as following diagnostic criteria of Southwest oncology group(SWOG) : plasmacytoma on tissue biopsy was noted 6 cases, bone marrow plasmacytosis with more than 10% plasma cells was 22 cases, monoclonal globulin spike on serum electrophoresis was 24 cases, lytic bone lesions was observed 22 cases. 4. Initial clinical stages were classified as 2 cases in stage I, 3 cases in stage II, 26 cases in stage III(84%) 5. Immunoelectrophoresis revealed the distribution of IgG 64%, light chain 22%, IgA 10%. Kappa to Lambda ratio of 1.1 : 1. 6. Hematologic & biochemical fingins revealed anemia with <8.5% of hemoglobulin in 42%, hypercalcemia with < 10.6mg% of serum calcium in 22%, azotemia >2.0mg% of serum creatinine in 19%. 7. The multiple punched out lesion of bone x-ray examination were noticed skull(65%), rib(42%), L-spine(35%), pelvis(23%), T-spine(19%). The initial skeletal roentgenographic findings showed osteoporosis, osteolytic lesion and fracture in 55%, only osteolytic lesion in 23%, only osteoporosis in 10%. 8. Complications of multiple myeoloma, such as 10 cases of renal impairment, 8 cases of infection, 16 cases of compression fracture of spine were observed.

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Vinblastine Determination Measured by a Sensitive ELISA Inhibition Assay (ELISA Inhibition Assay에 의한 Vinblastine의 측정)

  • Jae Wha kim;Mi Young Han;Hee Gu Lee;Eun Young Song;Tai Wha Chung;Kyung Soo Nam;In Seong Choe
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.121-126
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    • 1996
  • Specific monoclonal antibodies(mAbs) against bis-indole alkaloids related to vinblastine were established to develop a simple and specific immunoassay system for the quantitation of Vinca alkaloids. Vinca alkaloids were extracted from tissue cultured cells of periwinkle plant (Vinca rosea L.). Spleen cells from Balb/c mice immunized with vinblastine-bovine serum albumin(VBL-BSA) conjugate as immunogen were fused with myeloma cells(Sp2/0-Ag.14) in the presence of polyethylene glycol. In the preliminary experiments, 32 clones which highly reacted with VBL-BSA conjugate were selected by ELISA(Enzyme-linked immunosorbent assay).These clones were further analyzed by inhibition assay of ELISA. The results obtained with two typical monoclonal antibodies, KN-1 and KN-2, were described. KN-1 exhibited considerable reactivities with soluble dimeric bis-indole alkaloids, whereas no cross reacted with monomeric bis-indole alkaloids. However KN-2 showed cross reactivity with mono- and di-meric bis-indole alkaloids. Furthermore, KN-1 was applied to the immunoassay system for determining the VBL amounts of in vitro cultured cell extracts. This assay system could detect dimeric vinca alkaloid as low as 0.05 nM.

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The Development of Diagnosis for Atopic Dermatitis by Evaluating the Expression of Skin Barrier Proteins Using a Non-Invasive Method (비침습적 피부장벽단백질 측정을 통한 아토피 피부염의 진단 및 유용성)

  • Kim, In Sik;Lee, Ji-Sook
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.395-400
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    • 2017
  • Atopic dermatitis is a chronic inflammatory skin disease caused by a variety of genetic and environmental factors, dysregulation of immunological response, as well as dysfunction of the skin barrier proteins. The purpose of this study is to develop an ELISA kit suitable for evaluating the expression of skin barrier proteins. Proteins were obtained from the skin via AriNo and D-Squame patches. The efficiency of protein collection from the skin, using the Arino patch, was shown to be more effective than using D-Squame; while the efficiency of lysis using 0.1% Triton-X100 was higher than that of other lysis solutions, including 0.1 M Tris-HCL, 0.1% Tween-20, and 5 mM KOH. Recombinant skin barrier proteins, such as filaggrin and involucrin, were produced by molecular biological methods. Monoclonal antibodies against filaggrin and involucrin were produced by immunization of mice, fusion of spleen cells and myeloma cells, as well as a selection of antibody-producing hybridoma cells. The filaggrin expression in the skin of subjects suffering from atopic dermatitis was lower than that in normal mice. Involucrin expression was not altered between normal individuals and subjects with atopic dermatitis. These findings contribute to an elucidation of the importance of the skin barrier protein expression in atopic dermatitis and the development of a diagnostic kit for atopic dermatitis.

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.

White Sport Syndrome Virus Disease of Shirmp and Diagnostic Methods

  • Zhan, Wen-Bin
    • Journal of Aquaculture
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    • v.15 no.1
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    • pp.7-13
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    • 2002
  • Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.