• Title/Summary/Keyword: B3 antibody

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Production of monoclonal antibody(MAb) for lymphocytes of Korean native cattle 2. MAb to E-rosette forming lymphocytes. (한우 림프구에 대한 단일항체 생산 2. E-rosette형성 림프구에 대한 단일항체)

  • Song, Hee-jong;Woon, Jae-ho;Kim, Tae-joong
    • Korean Journal of Veterinary Research
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    • v.35 no.4
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    • pp.777-783
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    • 1995
  • A monoclonal antibody(MAb), KCT-23ER, with specificity for E-rosetted T cells of Korean native cattle was prepared by cell hybridization of myeloma P3/NS-1/1-Ag-4-1 and spleen cells from BALB/c mice hyperimmunized with E-rosetted lymphocytes. The isotype of KCT-23ER to T lymphocytes was mouse $IgG_{2b}$. KCT-23ER was reacted with 53.6% to peripheral blood lymphocytes and with 67.8% to nylon wool nonadherent blood lymphocytes. And it was reacted with 72.2%, 59.2% and 35.3% to thymocytes, prescapular lymph node cells and splenocytes, respectively. Immunocytological reactive rates to E-rosetted and non-E-rosetted cells were 72.5% and 22.4%, respectively. These results indicated that KCT-23ER reacted to E-rosetted cells was one of the MAb for investigate of $CD_2$ receptor positive cell subset in the Korean native cattle.

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The health status of stray dogs and cats adopted to public animal shelter in Gwangju area, Korea (광주지역 유기동물 건강 실태 조사)

  • Na, Ho-Myung;Choi, Jong-Woog;Park, Jae-Sung;Lee, Yeun-Ey;Bae, Seong-Yeol;Park, Seong-Do;Kim, Eun-Sun;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.37 no.4
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    • pp.281-290
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    • 2014
  • The purpose of this study was to determine the zoonotic diseases of stray dog and cat in Gwangju, Korea. We chose randomly 300 samples which is 265, dogs and 35, cats in the public animal shelter from March to August of 2013. The animals' blood biochemistry values measured, and the out of normal values were 49.7% GOT, 36.3% GPT, and 78.0% GGT. The tested items were Dirofilaria immitis, Toxoplasma gondii, Brucella canis, Rabies virus. The positive results were 10% Dirofilaria immitis, 6.3% Toxoplasma gondii (antibody detected), 10% Rabies (antibody detected) but 0.0% in B. canis. The stray animals' antibody antigen positivity take effect high from Mar.-May. Therefore, we propose that those diseases should be monitering and vaccinating in Korea.

Production of Recombinant Humanized Anti-HBsAg Fab Fragment from Pichia pastoris by Fermentation

  • Deng, Ning;Xiang, Junjian;Zhang, Qing;Xiong, Sheng;Chen, Wenyin;Rao, Guirong;Wang, Xunzhang
    • BMB Reports
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    • v.38 no.3
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    • pp.294-299
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    • 2005
  • In this report, we describe the high-yield secretory expression of the recombinant human anti-HBsAg Fab fragment from Pichia pastoris that was achieved by co-integration of the genes encoding the heavy and light chains (both under the control of alcohol oxidase promoter) into the genome of the yeast cells. The fed-batch fermentations were carried out in a 5 L scale. Both chains of the Fab were successfully expressed upon methanol induction. The absorbance ($OD_{600}$) of the broth can reach 350~500 at the end of fed-batch phase. After the induction, the expression level of the recombinant Fab (soluble) reached 420~458 mg/L. The recombinant Fab fragment was purified from the crude culture supernatant by ion exchange chromatography and the purity of the recombinant Fab fragment was over 95%. The affinity activities of the crude fermentation supernatant and the purified Fab were analyzed by indirect ELISA, which showed that the purified recombinant Fab fragment had high affinity activity with hepatitis B surface antigen.

Regulation of Interleukin-17 Production in Patients with Rheumatoid Arthritis by Phosphoinositide 3-kinase (PI3K)/Akt and Nuclear Factor KappaB (NF-κB) Dependent Signal Transduction Pathway (류마티스 관절염 환자의 말초혈액 단핵세포에서 Phosphoinositide 3-Kinase (PI3K)/Akt와 Nuclear Factor KappaB (NF-κB) 신호전달을 통한 IL-17 생성조절)

  • Kim, Kyoung-Woon;Cho, Mi-La;Lee, Sang-Heon;Min, So-Youn;Park, Mi Kyung;Park, Sung-Hwan;Jue, Dae-Myung;Kim, Ho-Youn
    • IMMUNE NETWORK
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    • v.3 no.4
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    • pp.310-319
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    • 2003
  • Inflammatory mediators has been recognized as an important role in the pathogenesis of rheumatoid arthritis (RA). IL-17 is increasingly recognized as an important regulator of immune and inflammatory responses, including induction of proinflammatory cytokines and osteoclastic bone resorption. Evidence of the expression and proinflammatory activity of IL-17 has been demonstrated in RA synovium and in animal models of RA. However, the signaling pathways that regulate IL-17 production remain unknown. In the present study, we investigated the role of the phosphatidylinositol 3 kinase (PI3K)-Akt pathway in the regulation of IL-17 production in RA. PBMC were separated from RA (n=24) patients, and stimulated with various agents (anti CD3, anti CD28, PHA, ConA, IL-15). IL-17 levels were determined by sandwich ELISA and RT-PCR. The production of IL-17 was significantly increased in cells treated with anti-CD3 antibody, PHA, IL-15 or MCP-1 (P<0.05). ConA also strongly induced IL-17 production (P<0.001), whereas TNF-alpha, IL-1beta, IL-18 or TGF-beta did not. IL-17 was detected in the PBMC of patients with osteoarthritis (OA) but their expression levels were much lower than those of RA PBMC. Anti-CD3 antibody activated the PI3K-Akt pathway and activation of the PI3K-Akt pathway resulted in a pronounced augmentation of nuclear factor kappaB ($NF-{\kappa}B$). IL-17 production by activated PBMC in RA is completely or partially blocked in the presence of $NF-{\kappa}B$ inhibitor PDTC and PI3K-Akt inhibitor, wortmannin and LY294002, respectively. Whereas the inhibition of AP-1 and extracellular signal-regulated kinase (ERK)1/2 did not affect IL-17 production. These results provide new insight into that PI3K/Akt and $NF-{\kappa}B$ dependent signal transduction pathway could be involved in the overproduction of key inflammatory cytokine, IL-17 in rheumatoid arthritis.

Effects of Fermented Milk with Hot Water Extract from Acanthopanax senticosus and Codonopsis lanceolata on the Immune Status of Mouse (가시오가피와 더덕 추출물을 첨가한 발효유가 마우스의 면역기능에 미치는 영향)

  • Lim, Sang-Dong;Seong, Ki-Seung;Kim, Kee-Sung;Han, Dong-Un
    • Korean Journal of Food Science and Technology
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    • v.39 no.3
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    • pp.323-329
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    • 2007
  • We investigated the immunomodulatory actions of water extract from Acanthopanax senticosus in male ICR mice. The mice were treated with fermented milk containing three added doses of freeze dried extract: 3 mg/kg (A), 9 mg/kg (B), and 27 mg/kg (C) of body weight with Acanthopanax senticosus: Codonopsis lanceolata (8:2) for 7 and 10 weeks, respectively. Organ weights, plaque-forming cell tests, agglutination tests, IgG tests, differential white cell counts, and histological tests were performed at the 7th and 10th weeks of dietary treatment. There were no significant differences in body weight and organ weight. The spleen indices of group B at 7 weeks and group C at 10 weeks were significantly higher than those of the control group (p<0.05). For the plaque-forming cell test, groups B and C at 7 weeks, and group C at 10 weeks, showed significant increases over the control group (p<0.05). The agglutination test decreased with an extended experimental period. Groups A, B, and C at 7 weeks, and groups B and C at 10 weeks, had greater antibody responses to sheep red blood cells (SRBC) than the control group. The IgG antibody production of group C at 7 weeks and groups B and C at 10 weeks were significantly higher than the control group (p<0.05). In groups B and C, lymphocyte percentage was higher than the control group, and their spleen and thymus tissues showed active immune reactions.

Development of Chemiluminescence Immunoassay For the Measurement of Serum Thyroxine(T4)

  • Kim, J.B.;Choe, B.K.;Choi, S.H.
    • Korean Journal of Animal Reproduction
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    • v.11 no.1
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    • pp.16-21
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    • 1987
  • We describe a simple, solid-phase chemiluminescence immunoassay for the meausrement of serum T4. An immunoglobulin G fraction of antibody to thyroxine was passively absorbed onto the walls of polystyrene tubes. The labeled antigen was thyroxine-aminobutylethylisoluminol. After the bindings reaction (37$^{\circ}C$ for 1 hour), the solution is removed by aspiration and the antibody-bound fraction was washed once with buffer. Sodium hydroxide (5mol/1,200${mu}ell$) was added and the mixture incubated for 30 minutes at 6$0^{\circ}C$. Luminescence was initiated by oxidation of the label with micropeeroxidase-hydrogen peroxide and the signal of light emission was intergrated for 10 sec. The light yield was inversely proportional to the concentration of T4 in the standard or sample. An evaluation of the method gave the following values sensitivity of calibration curve 7.5$\pm$2.8 nmol/l (mean$\pm$SD). The intra-assay precision (CV%) was 8.9, 7.3 and 5.4. The inter-assay precision (CV%) was 10.2, 8.1 and 7.1. When seum samples were assayed for T4, the results obtained by solid-phase CIA and the conventional RIA agreed well(n=3.5, r=0.954).

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Purification of a Mosquitocidal Toxic Protein from B. thuringiensis strain H9B by Immuno-Affinity Chromatography (Immuno-Affinity Chromatography에 의한 B. thuringiensis H9B 균주의 모기살충성 내독소 단백질의 정제)

  • 김광현;배수장;이광배
    • Journal of environmental and Sanitary engineering
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    • v.12 no.2
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    • pp.59-64
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    • 1997
  • For purification of a 70kDa toxic protein of mosquitocidal delta-endotoxin from B. thuringiensis strain H9B, immuno-affinity chromatography was performed. After separation of 70kDa toxic proteins from the delta-endotoxin of the strain H9B on SDS-PAGE, the 70kDa toxic protein was subcutaneously injected into rabbit for making a polyclonal antibody. A anti-70kDa toxic protein was purified by a column chromatography packed with protein A-sepharose 4B gels. The 70kDa toxic protein from delta-endotoxin of the strain H9B was also purified by an immuno-affinity chromatography packed with CNBr-activated sepharose 4B gels conjugated anti-70kDa toxic protein after elution with 1/10M citric acid-1/5M Na$_{2}$HPO$_{4}$ buffer(pH3.2) containing 0.5M NaCl. The 70kDa toxic protein was purified through only one step-separation system, was demonstrated by SDS-PAGE and immunoblot.

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Effects of Glucose and Glutamine Concentrations on Hybridoma Growth ann Anti-HBsAg MAb Production in Enriched Medium (농축배지에서 Glucose와 Glutamine 농도가 하이브리도마 증식과 간염표면항원에 대한 단일클론항체 생산에 미치는 영향)

  • 전복환;조의철김동일
    • KSBB Journal
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    • v.5 no.4
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    • pp.365-371
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    • 1990
  • To improve the growth of mouse hybridoma 2c3.1 secreting anti-Hepatitis B surface antigen monoclonal antibody (anti-HBsAg MAb), we had constructed an enriched medium and observed the effects of fetal bovine serum and serum-free supplements including human serum albumin, 'insulin and transferrin', and monoethanolamine. For further enhancement of growth, the concentrations of two major energy sources, glucose and glutamine, were strengthened with various ratios in the enriched medium. Maximum cell growth and monoclonal antibody production obtained in various ratios of glucose/glutamine with an inoculation concentration of 2$\times$105 cells/ml were 0.73$\times$106-4.62$\times$106 cells/ml and 65.1-422.6 $\mu\textrm{g}$/ml, respectively. Glutamine was round to be a major energy source and a limiting nutrient in comparison to glucose for 2c3.1 cell cultivation in enriched media with low serum.

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Agonistic Anti-CD137 Monoclonal Antibody Treatment Induces CD11b+Gr-1+ Myeloid-derived Suppressor Cells

  • Lee, Jung-Mi;Seo, Jeong-Hwan;Kim, Yeon-Jeong;Kim, Yun-Sun;Ko, Hyun-Jeong;Kang, Chang-Yuil
    • IMMUNE NETWORK
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    • v.10 no.3
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    • pp.104-108
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    • 2010
  • CD137 (4-1BB/tnfrsf9) has been shown to co-stimulate T cells. However, agonistic anti-CD137 monoclonal antibody (mAb) treatment can suppress $CD4^+$ T cells, ameliorating autoimmune diseases, whereas it induces activation of $CD8^+$ T cells, resulting in diverse therapeutic activity in cancer, viral infection. To investigate the CD137-mediated T cell suppression mechanism, we examined whether anti-CD137 mAb treatment could affect $CD11b^+Gr-1^+$ myeloid-derived suppressor cells (MDSCs). Intriguingly, anti-CD137 mAb injection significantly increased $CD11b^+Gr-1^+$ cells, peaking at days 5 to 10 and continuing for at least 25 days. Furthermore, this cell population could suppress both $CD8^+$ T cells and $CD4^+$ T cells. Thus, this study demonstrated that, for the first time, anti-CD137 mAb treatment could induce $CD11b^+Gr-1^+$ MDSCs under normal conditions, suggesting a possible relationship between myeloid cell induction and CD137-mediated immune suppression.