• Title/Summary/Keyword: antibody formation

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Effects of the Glycoprotein Isolated from Pteridium aquilinum on the Immune Function of Mice (고사리 단백다당(Pteridium aquilinum Glycoprotein, PAG)이 마우스 면역활성에 미치는 영향)

  • Park, Hyeon-Ae;Kweon, Mee-Hyang;Han, Hyung-Mee;Sung, Ha-Chin;Yang, Han-Chul
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.976-982
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    • 1998
  • The effects of the glycoprotein (PAG) isolated from Pteridium aquilinum on the immune function was examined in mice. PAG was intraperitoneally administered into BALB/C mice for 14 days and the antibody forming ability to hen egg lysozyme (HEL) and the blastogenic responses of splenocytes were measured. PAG treatment significantly increased antibody formation to HEL in a dose-dependent manner. Blatogenesis of splenocytes in response to lipopolysaccharide (LPS, B-cell specific mitogen) or phytohemagglutinin (PHA, T-cell specific mitogen) was also increased after treatment with PAG, indicating that the PAG increases both humoral and cellular immunities. To examine whether the immune function of PAG was via a direct effect on the lymphocytes, splenocytes were isolated from BALB/C mice, exposed to various concentrations of PAG in vitro and the blastogenic responses were measured. In vitro exposure to PAG significantly increased blastogenesis of splenocytes to LPS up to $500{\;}{\mu}g/kg$, whereas the blastogenic response to PHA was not altered by PAG treatment. To identify the fraction responsible for the increase in the immune function, the effect of periodate digest, pronase digest or purified polysaccharide on the antibody production to HEL was examined. Crude protein fraction of PAG significantly increased the antibody formation to HEL. On the other hand, both crude and purified polysaccharide fractions did not have any effects on the antibody production ability. These data indicated that 1) PAG increased both humoral and cellular immune functions, 2) the increase in humoral immunity was probably via a direct action of PAG on lymphocytes and 3) the protein portion of PAG was responsible for the increase in humoral immunity.

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Fucoidan Increases Porcine Neutrophil Extracellular Trap Formation through TNF-α from Peripheral Blood Mononuclear Cells

  • Changwoo Nahm;Yoonhoi Koo;Taesik Yun;Hakhyun Kim;Byeong-Teck Kang;Mhan-Pyo Yang
    • Journal of Veterinary Clinics
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    • v.40 no.3
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    • pp.175-181
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    • 2023
  • Fucoidan extracted from brown seaweed has a variety of biological activities. Neutrophil extracellular traps (NETs) formation is an immune response for the invasion of pathogens. Neutrophils release granule protein and chromatin that form extracellular fibers that bind microbes. These NETs degrade virulence factors and kill bacteria. The aim of this study was to investigate the effect of fucoidan on NET formation of porcine peripheral blood polymorphonuclear cells (PMNs). The NET formation was determined by fluorescence emission of propidium iodide (PI) in PMNs by a fluorescence microplate reader. The production of tumor necrosis factor (TNF)-α from peripheral blood mononuclear cells (PBMCs) was measured by ELISA method. Fucoidan itself did not show any direct effect on NET formation. However, NET formation of PMNs was increased by the culture supernatant from PBMCs treated with fucoidan. The NET formation of PMNs were also enhanced by treatment with recombinant porcine (rp) TNF-α. The ability of culture supernatant from PBMCs treated with fucoidan to increase the NET formation of PMNs was inhibited by addition of goat anti-rp TNF-α polyclonal antibody (pAb) (IgG) prior to the culture. The increase of NET formation by rp TNF-α was also inhibited by goat anti-rp TNF-α pAb (IgG). The level of TNF-α in culture supernatant from PBMCs was increased by treatment with fucoidan. These results suggest that fucoidan increases porcine NET formation, which is mediated by TNF-α produced from PBMCs.

Proteasome Inhibitors Affect Appressorium Formation and Pathogenicity of the Rice Blast Fungus, Magnaporthe oryzae

  • Wang, Yiming;Kim, Sang-Gon;Wu, Jingni;Yu, Seok;Kang, Kyu-Young;Kim, Sun-Tae
    • The Plant Pathology Journal
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    • v.27 no.3
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    • pp.225-231
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    • 2011
  • Previously, we identified the 20S proteasome ${\alpha}$-subunit of Magnaporthe oryzae (M. oryzae) induced during appressorium formation, and detected an increase in multiple protein ubiquitination during the early appressorium formation process (Kim et al., 2004). In this study, we further attempted to determine whether the proteasome is involved in the appressorium formation of M. oryzae both in vitro and in planta, using proteasome inhibitors. A significant increase in 20S proteasome during fungal germination and appressorium formation was observed using Western blot analysis with 20S proteasome antibody, demonstrating that proteasome-mediated protein degradation was involved in appressorium formation. Pharmacological analysis using proteasome inhibitors, MG-132, proteasome inhibitor I (PI) and proteasome inhibitor II (PII) revealed that germination and appressorium formation were delayed for 4 to 6 h on rice leaf wax-coated plates. Similarly, the treatment of proteasome inhibitors with fungal conidia on the rice leaf surface delayed appressorium formation and host infection processes as well. Additionally, fungal pathogenicity was strongly reduced at 4 days' postfungal infection. These data indicated that the fungal 20S proteasome might be involved in the pathogenicity of M. oryzae by the suppression of germination and appressorium formation.

Zinc Enhances Neutrophil Extracellular Trap Formation of Porcine Peripheral Blood Polymorphonuclear Cells through Tumor Necrosis Factor-Alpha from Peripheral Blood Mononuclear Cells

  • Heo, Ju-Haeng;Kim, Hakhyun;Kang, Byeong-Teck;Yang, Mhan-Pyo
    • Journal of Veterinary Clinics
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    • v.37 no.5
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    • pp.249-254
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    • 2020
  • Neutrophil extracellular trap (NET) formation is an immune response for the invasion of microbes. The purpose of this study is to examine the effect of zinc on NET formation of porcine peripheral blood polymorphonuclear cells (PMNs). The NET formation of PMNs was measured by fluorescence microplate reader. The production of tumor necrosis factor (TNF)-α in the culture supernatants from zinc-treated peripheral blood mononuclear cells (PBMCs) was measured by enzyme-linked immunosorbent assay (ELISA). Zinc itself did not have no effect on NET formation. However, the NET formation of PMNs was increased by culture supernatants from PBMCs treated with zinc. Also, the NET formation of PMNs was increased by recombinant porcine (rp) TNF-α. The production of TNF-α in PBMCs culture supernatants was shown to increase upon zinc treatments. These NET formations of PMNs increased by either culture supernatant from PBMCs treated with zinc or rpTNF-α were inhibited by treatment of anti-rpTNF-α polyclonal antibody (pAb). These results suggested that zinc has an immunostimulating effect on the NET formation of PMNs, which is mediated by TNF-α released from zinc-treated PBMCs. Therefore, zinc may play an important role for NET formation in the defense of porcine inflammatory diseases.

Antibody Producibilities of Salmonella typhi in Mice fed on Different Fatty Acids (지방산을 투여한 마우스의 Salmonella typhi에 대한 항체 생성력)

  • 이정화;김용호;이원재;함건주
    • Biomedical Science Letters
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    • v.1 no.1
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    • pp.45-54
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    • 1995
  • The effect of different fatty acids supplementation on antobody production of Salmonella typhi was studied in ICR mice. Subjects supplemented their diets with $50\mu$g of extracted pig oil(as a saturated fatty acid) and fish oil (as a unsaturated fatty acid) / 2 days for 8 weeks. Blood was collected control and experimental groups of mice after 8 weeks of oil supplementation. The different fatty acids supplementation reduced unsaturated fatty acids composition in mice liver such as $C_{18:3}, \; C_{20:3}\; and\; C_{20:4}\; except\; C_{18:1}\; and\; C_{18:2}/C_{18:0}$ in fish oil and pig oil groups compared to control group. Also, the phagocytic activities of mice macrophages for Candida albicans was reduced by 6% in pig oil group and 9% in fish oil group than control group. The antigen-stmulated lympocite proliferative response was significantly increased by fatty acid in pig oil group(48%) but 57% in fish oil group. The different fatty acid supplementation increased antibody production in both experimental groups than control group ; this increase was only significant in pig oil group(1:$2^4$) on mice but not in fish oil group(1:$2^0$) compared to control group(1:$2^0$), however, increased antibody titer in both groups in vitro spleen cell culture supernatant(1:$2^3$ in fish oil group and 1:$2^2$ in pig oil group compared to control group 1:$2^0$). Thus, fish oil supplementation was immunosuppresive agent in macrophage phagocytosis, in-vivo antobody producibilities and lympocyte proliferation but pig oil supplementation was more effective than fish oil in antibody formation in-vivo. We find that antibody producibilities affected by fed on different fatty acids were considered by balance between saturated and unsaturated fatty acid, and $C_{20:3}/C_{20:4}$ ratio. Also, it affected to antigen-stimulated lymphocyte proliferation and macrophage phagocytic activities.

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Effect of specific serum IgG antibody against Streptococcus mutans on the adherence of S. mutans to smooth surface in vitro (특이혈청항체(特異血淸抗體) IgG분획(分劃)이 Streptococcus mutans의 평활면(平滑面) 부착(附着)에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Lee, Jean-Yong;Choi, Eu-Gene;Ha, Youn-Mun;Kim, Chan-Soo
    • The Journal of the Korean Society for Microbiology
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    • v.17 no.1
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    • pp.75-85
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    • 1982
  • In order to demonstrate the effect of specific serum IgG antibody on the adherence of Streptococcus mutans to smooth surface and the mechanism of effective adherence inhibition by IgG antibody, in the present study authors obtained purified IgG from different immunogen preparations of S. mutans NCTC 10449(serotype c) and observed the effect of each IgG preparation on the adherence of each S. mutans strain cultured in different conditions. In addition, the present study was undertaken to observe the cross-reactivity of IgG and the effect of sucrose concentration on the adherence of S. mutans in vitro non-growth condition. The adherence of S. mutans to glass surface was effectively inhibited by serum IgG antibody. At the same IgG concentrations, anti-2% fructose grown/1N NaCl washed S. mutans NCTC 10449 cell showed greater adherence inhibitory effect to S. mutans strains than anti-2% sucrose grown and anti-S. mutans NCTC 10449 cell wall, and the greater inhibitory effects of IgG preparations were observed in assay using 2% fructose grown S. mutans cell preparations than using 0.1% sucrose grown cell preparations. These results suggest that the more effective adherence inhibition by serum IgG antibody is due to the reaction with S. mutans cell surface antigens rather than glucan and cell-associated glucosyltransferase. The greatest adherence inhibitory effect of IgG to S. mutans strains was observed on homologous NCTC 10449 strain and the inhibition cross-reactivities were observed between serotype c, e, and f strains. More pronounced cross-reactivity of adherence inhibition of IgG to S. mutans was observed in assay using anti-2% fructose grown/1N NaCl washed cell than using other IgG preparations, and observed in assay using 2% fructose grown S. mutans cell preparations than 0.1% sucrose grown cell preparations. It was interested that low, but adequate concentration of reactive IgG antibody significantly increased the adherence ability of S. mutans. This result may be due to the formation of small cell aggregates resulted in a increase in the numbers of organisms which adhered to glass surface. The adherence of S. mutans to glass surface was possible in the absence of glucan-synthetic activity. Low level of sucrose significantly increased the adherence ability of S. mutans to glass surface, but excessive amount of sucrose induced large cell aggregates resulted in a decrease in the numbers of organism which adhered.

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Effect of Reactive Oxygen Species on the Formation of Nε-(carboxymethyl)lysine, an Endproduct of Maillard Reaction of Proteins (단백질의 Maillard 반응의 최종산물인 CML 형성에 미치는 ROS의 영향)

  • Lee, Jong-Sun;Yang, Ryung;Shin, Dong-Bum
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.1
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    • pp.123-131
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    • 2004
  • The effect of reactive oxygen species (ROS) on the formation of $N^{\varepsilon}$-(carboxymethly)lysine (CML). one of the endproducts in the Maillard reaction of protein (or glycation), was investigated. Glyoxal, a main precursor of CML formation, was produced from both glucose and fructose during their autoxidation. The transition metal ion showed to involve in the formation of glyoxal by the metal catalyzed oxidation, suggesting that ROS accelerated the reducing sugar autoxidation. The stimulative effect of ROS on the autoxidation was more prominent in glucose than in fructose. Polyunsaturated acids (PUFAs) were shown to form glyoxal by peroxidation in proportion to the degree of unsaturation, but ROS did not affect on PUFA peroxidation. Ascorbic acid also lysine (CMHL) in the model system using hippuryl lysine and glucose had a significant effect on ROS, whereas it had no effect on ROS using glyoxal as a reactant. Almost the same trend was obtained by the analysis of antigen coated indirect noncompetitive ELISA using monoclonal antibody (6D12). These data indicated that ROS affected glucose autoxidation as well as mediated both CML and glyoxal formation, but did not affect the reactive compounds such as fructose, PUFAs and ascorbic acid.

Stability of $^{188}Re$ Labeled Antibody for Radioimmunotherapy and the Effect of Stabilizing Agents (방사면역치료용 $^{188}Re$ 표지 항체의 안정성과 안정제의 효과)

  • Chang, Young-Soo;Kim, Bo-Kwang;Jeong, Jae-Min;Chung, June-Key;Lee, Seung-Jin;Lee, Dong-Soo;Lee, Myung-Chul
    • The Korean Journal of Nuclear Medicine
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    • v.36 no.3
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    • pp.195-202
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    • 2002
  • Purpose: For clinical application of beta-emitter labeled antibody, high specific activity is imporiant. Carrier-free $^{188}Re$ from $^{188}W/^{188}Re$ generator is an ideal radionuclide for this purpose. However, low stability of $^{188}Re$ labeled antibody, especially in high specific activity, due to radiolytic decomposition by high energy (2.1 MeV) beta ray was problem. We studied the stability of $^{188}Re$ labeled antibody, and stabilizing effect of several stabilizers. Materials and Methods: Pre-reduced monoclonal antibody (CEA79.4) was labeled with $^{188}Re$ by incubating with generator-eluted $^{188}Re-perrhenate$ in the presence of stannous tartrate for 2 hr at room temperature. Radiochemical purity of each preparation was determined by chromatography. Human serum albumin was added to the labeled antibodies (2%). Stability of $^{188}Re-CEA79.4$ was investigated in the presence of ascorbic acid, ethanol, of Tween 80 as stabilizing agents. Results: Labeling efficiencies were $88{\pm}4%\;(n=12)$. Specific activities of $1.25{\sim}4.77MBq/{\mu}g$ were obtained. If stored after purging with $N_2$, all the preparations were stable for 10 hr. However, stability decreased in the presence of air. Perrhenate and $^{188}Re-tartrate$ was major impurity in declined preparation. colloid-formation was not a significant problem in all cases. Addition of ascorbic acid stabilized the labeled antibodies either under $N_2$ or under air by reducing the formation of perrhenate. Conclusion: High specific activity $^{188}Re$ labeled antibody is unstable, especially, in the presence of oxygen. Addition of ascorbic acid increased the stability.

Expression and Localization of Brain Glutamate Dehydrogenase with Its Monoclonal Antibody

  • Lee, Jong-Eun;Choe, Su-Yeong;Jo, Seong-U
    • Animal cells and systems
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    • v.2 no.1
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    • pp.71-80
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    • 1998
  • Glutamate dehydrogenase (GDH) is one of the main enzymes involved in the formation and metabolism of the neurotransmitter glutamate. In the present study, we investigated the distribution of the GDH-immunoreactive cells in the rat brain using monoclonal antibodies against bovine brain GDH isoprotein. GDH-immunoreactive cell were distributed in the basal ganglia, thalamus and the nuclei belong to substantia innominata, and its connecting area, subthalamic nucleus, zona incerta, and substantia niqra. We could see GDH-immunoreactive cells in the hippocampus, septal nuclei associated with the limbic system, the anterior thalamic nuclei connecting between the hypothalamus and limbic system, and its associated structures, amygdaloid nuclear complex, the dorsal raphe and median raphe nuclei and the reticular formation of the midbrain. The GDH-immunoreactive cells were shown in the pyramidal neurons of the cerebral cortex, the Purkinie cells of the cerebella cortex, their associated structures, ventral thalamic nuclei and the reticular thalamic nuclei that seem to function as neural conduction in the thalamus.

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Influence for Carcinoma Cell and Lymphatic Cell of Acetyl Arsonate (아세틸 아르소네이트의 면역세포와 암세포에 미치는 영향: L1210, Sarcoma 180, MOLT-4 등 사람과 생쥐에 대한 작용비교)

  • Jeong, Yong-Ja;Seong, Yeong-Gi
    • YAKHAK HOEJI
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    • v.40 no.5
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    • pp.599-607
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    • 1996
  • Acetylarsonate was prepared for testing antitumor and immunological effects. It showed cytotoxicity directly on Sarcoma 180. L1210 and MOLT-4 by MTT assay. It did not seemed to trigger the mitosis of human lymphocytes in culture, but that showed the cytotoxicity with higher dose. The rosette formation and spleen weight of mouse which acetylarsonate was administered to for 2 weeks were increased. Furthermore, peripheral helper T- and cytotoxic/suppressor T-lymphocytes were increased in acetylarsonate-injected-mice significantly when it was estimated with simultaneous 2 color analysis using anti Lyt2-FITC and L3T4-PE monoclonal antibody by Flow cytometer.

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