• Title/Summary/Keyword: 세포성면역반응

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Export of Human Proinsulin in E. coli : High Export of Proinsulin Fusion Protein but not of Proinsulin Itself (대장균에서 인체 프로인슐린의 분비 발현 : 프로인슐린 융합체의 고분비 발현과 프로인슐린의 저분비 발현)

  • Yup Kang
    • KSBB Journal
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    • v.11 no.2
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    • pp.165-172
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    • 1996
  • To obtain a correctly folded human proinsulin, export of proinsulin using Staphylococcal protein A signal sequence-mediated secretion pathway has been attempted in E.coli. A secretion operon for proinsulin was constructed by consecutively connecting T7 promoter, SPA ribosome binding site, SPA signal sequence gene, and human proinsulin gene. Little immunoreactive proinsulin was detected in the periplasmic space and. culture medium, and not even in cytoplasmic space. The qualitative analysis of transcribed proinsulin mRNA and the in vitro transcription/translation experiment suggests that the negligible level of proinsulin export appears to be due to intracellular degradation of proinsulin, rather than due to the blockage during translocation. However, expression of proinsulin fusion protein such as MBP-proinsulin could dramatically increase export of proinsulin in E.coli.

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Studies on the Effect of Corilagin Isolated from Euphorbia helioscopia on Collagen-Induced Arthritis (II);CytokinesAnalysis of Collagen II Induced Arthritis Mice Model (택칠에서 분리한 Corilagin이 Collagen 유발 관절염에 미치는 영향 (II);Corilagin을 투여한 류마티스 관절염 유발 생쥐의 사이토카인 분석)

  • Shin, Sam-Kee;Chang, Jun-Pok;Doh, Eun-Soo
    • Korean Journal of Plant Resources
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    • v.21 no.4
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    • pp.336-340
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    • 2008
  • Corilagin (CRN) isolated from Euphorbia helioscopia as rheumatoid arthritis drug. CRN was medicated to the abdominal cavity of collagen induced arthritis (CIA) mice that was an animal model for rheumatoid arthritis and its effects on incidence and arthritis index were studied. The results were as folllows; It was exhibited that medicating corilagin inhibited the infiltration of activated T lymphocytes into an inflammatory joint. The production of IgG and IgM that were RF (rheumatoid factor) and inflammatory cytokine, IL-6 and $TNF-{\alpha}$were reduced. After measuring $IFN-{\gamma}$and IL-4, it was found that it was shifted into Th2 immune response as increasing in IL-4. After liver function test, studies on liver poisoning of AST/ALT should be continued.

Study of Deep Ground Sea-Like Water on Antioxidant Activity and the Immune Response in RAW264.7 Macrophages. (천연 암반 심해수의 항산화 활성 및 면역반응에 대한 연구)

  • Kim, Yu-Jung;Jung, Il-Sun;Song, Hyo-Ju;Choi, Eun-Young;Choi, In-Soon;Choi, Young-Ju
    • Journal of Life Science
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    • v.18 no.3
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    • pp.329-335
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    • 2008
  • Korean Deep ground sea-like water (KDSW) has a similar mineral composition with deep sea water. KDSW has demonstrated its usefulness and attracted in the medical fields. KDSW and Danasoo (desalted deep ground sea-like water) intake improve antioxidant, antidiabetic activity and immunity. Antioxidant activities of KDSW and Dnansoo were measured by using 2,2-diphenyl-l-picryl-hydrazyl (DPPH) free radical, superoxide dismutase-like activity (SODA) and photochemiluminescence (PCL). DPPH radical scavenging and SOD-like activities of KDSW and Danasoo were remarkably increased in a dose-dependent manner. Antioxidant activities of KDSW and Danasoo 85.32 and 14.02 nmol of ascorbic acid equivalent/ml KDSW and Danasoo, respectively, using the PCL method. Lipopolysaccharide (LPS)-induced nitric oxide (NO) production in macrophages RAW264.7 cells was inhibited up to 30% by treatment with Danasoo (20%). NO is synthesized by the enzyme of nitric oxide synthase (NOS) and plays an important role tumor growth and angiogenesis. The anticancer effects of Danasoo on human gastric and lung cancer cells was performed by levels of inducible nitric oxide synthase (iNOS). Danasoo significantly reduced iNOS expression of human gastric cancer (SNU-l) and lung carcinoma (A549). The serum glucose level was significantly reduced by Danasoo (20%) diet in streptozotocin (STZ)-induced diabetic rats. These result suggest that KDSW has excellent biological activities and thus it has great potential as a source for natural health products.

Biological Roles of the Glycan in the Investigation of the Novel Disease Diagnosis and Treatment Methods (신개념 질병 진단 및 치료 연구에 있어서의 당사슬의 생물학적 역할)

  • Kim, Dong-Chan
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1379-1385
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    • 2018
  • Glycans are attached to proteins as in glycoproteins and proteoglycans. They are found on the exterior surface of cells. O- and N-linked glycans are very common in eukaryotic cells but may also be found in prokaryotes. The interaction of cell surface glycans with complementary glycan binding proteins located on neighboring cells, other cell types, pathogens like virus, or bacteria is crucial in biologically and biomedically important processes like pathogen recognition, cell migration, cell-cell adhesion, development, and infection. Their implication in pathological condition, suggests an important role for glycans as disease markers. In addition, a great amount of research has been shown that appropriate glycosylation of a recombinant therapeutic protein is critical for product solubility, stability, pharmacokinetics and pharmacodynamics, bioactivity, and safety. Besides, cancer-associated glycosylation changes often involve sialic acid in glycan branch which play important roles in cell-cell interaction, recognition and immunological response. This review aims at giving a comprehensive overview of the glycan's biological function and describing the relevance among the glycosylation, disease diagnosis and treatment methods. Furthermore, the high-throughput analytic methods available to measure the profile changing patterns of glycan in the blood serum as well as possible underlying biochemical mechanisms.

The Preventive and Therapeutic Effects of Probiotics in Allergic Diseases Via Immune Modulation (프로바이오틱스의 면역조절을 통한 알레르기 예방 및 치료효과)

  • Kim, Yeon-Hui;Choi, Chang-Yong;Chun, Taehoon
    • Journal of Food Hygiene and Safety
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    • v.31 no.3
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    • pp.141-152
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    • 2016
  • The demand to develop more safe and efficient methods for treating allergic patient is now continuously growing due to the increasing prevalence of allergic diseases. Probiotics are endogenous microbial flora that gives health benefits within hosts. Probiotics are now considered as one of solutions to treat allergic patients since recent evidence shows that some of probiotics have immunomodulatory function. Also, the treatment of probiotics to patients is relatively safer than other anti-inflammatory agents. In this review, we summarized on immunomodulatory function of some probiotics which show preventive or therapeutic effects on major allergic diseases such as atopic dermatitis, allergic rhinitis, asthma, or food allergy. Based on previous literature, the treatment of probiotics can alleviate the symptoms of allergic diseases via balancing $Th_1/Th_2$ response or increasing the number of regulatory T ($T_{reg}$) cells.

Radiolabeling and Immunological Characteristics of In-house Anti-Leukemic Monoclonal Antibodies(Anti-CALLA, Anti-JL-1 Antibodies) (국산 항 백혈병 항체(항 CALLA, 항 JL-1)의 동위원소 표지 및 면역학적 특성에 관한 연구)

  • So, Young;Chung, June-Key;Jeong, Jae-Min;Lee, Dong-Soo;Lee, Myung-Chul;Koh, Chang-Soon;Park, Seong-Hoe
    • The Korean Journal of Nuclear Medicine
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    • v.29 no.1
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    • pp.98-104
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    • 1995
  • Recently murine monoclonal antibodies have been studied actively for radioimmuno-scintigraphy and radioimmunotherapy, especially on patients with leukemia and lymphoma. In this research, we studied radiolabeling and immunologic characteristics of two in-house anti-leukemic monoclonal antibodies(anti-CALLA & anti-JL-1 antibodies) to make the basis for their clinical application. Each antibody was radiolabeled successfully with $^{99m}Tc$ by pretargeting transchelation method and with $^{125}I$ by lodogen method. We also studied cell binding assay, Scatchard analysis and modulation phenomenon. $^{125}I$ showed 90% labeling efficiency for each anti-body which was satisfactory, but $^{99m}Tc$ showed labeling efficiency below 70%, for which we need better labeling method. In cell binding assay, the immunoreactivity(IR) was low for $^{99m}Tc$-labeled antibodies. Scatchard analysis showed satisfactory data for both binding affinity. The affinity constant and antibody binding sites per cell are around $10^9M^{-1}$ and $10^4$, respectively. There was no modulation phenomenon in cases of $^{125}I$ or $^{99m}Tc$ labeled antibodies. We expect that two anti-leukemic monoclonal antibodies may be useful in diagnosis and therapy for leukemia and lymphoma patients.

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Production of a Monoclonal Antibody to Human $\alpha$-Fetopotein and Development of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assays for Human $\alpha$-Fetoprotein (인간 $\alpha$-fetoprotein에 대한 모노클로날 항체의 제조 및 모노클로날 항체를 이용한 효소면역분석법의 개발)

  • Michung Yoon;Hyun-Hee Lee;Youngwon Lee
    • Biomedical Science Letters
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    • v.5 no.1
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    • pp.1-10
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    • 1999
  • This study was attempted to generate a monoclonal antibody against human $\alpha$-fetoprotein (AFP) and to produce an immunoassay, recognizing AFP in plasma and amniotic fluid. AFP was purified from human amniotic fluid and used to immunize mice. Spleens were taken from the mice and the cells were fused with mouse myeloma cells (Sp2/0-Ag-14) for the production of monoclonal antibodies by employing the hybridoma technology. As a result, a hybridoma cell line producing anti-AFP monoclonal antibody was cloned out and designated as MabF22. From isotyping analysis, it was found that monoclonal antibody MabF22 was IgG type with IgG1 heavy chain and k light chain. The binding specificity of MabF22 was analyzed by immunoblotting as well as by ELISA. MabF22 was highly specific, reacting with only AFP-containing samples. The binding affinity was determined by ELISA (free-capture mode) and Scatchard analysis. As a result, the value of Kd was 0.8$\times$10$^{-10}$M. The validity of the MabF22 for AFP assay was examined by two kinds of ELISAs, i.e., non-competitive and competitive ELISA. Both assays revealed that MabF22 reacted well with AFP in sample in a concentration-dependent manner. Standard curve and antibody titration curve were obtained by using purified AFP and MabF22. These results indicate that the monoclonal antibody produced in this study would be useful not only for research purposes but also for further development of immune-diagnostic kit for the measurement of AEP concentration.

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Regulatory Mechanism of Lipopolysaccharide(LPS)-Induced Interleukin-8 Gene Expression in Mononuclear Phagocytic Cells (단핵식세포에서 내독소에 의한 인터루킨-8 유전자 발현 조절기전에 관한 연구)

  • Yoo, Chul-Gyu;Suh, Gee-Young;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.5
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    • pp.462-474
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    • 1994
  • Background : In acute lung injury, activated neutrophils play an important role in tissue damage. For neutrophils to participate in lung inflammation, chemotactic factors released from mononuclear phagocytes are needed to bring these cells to the local site of inflammation, with interleukin-8 (IL-8) being one of the most specific and important chemotactic factors for neutrophils. IL-8 also induces the expression of adhesion molecules and activates neutrophils to release various inflammatory mediators. Lipopolysaccharide(LPS) is one of the most important causes of adult respiratory distress syndrome and can cause release of many inflammatory cytokines including IL-8 leading to acute lung injury. But little is known about the regulatory mechanism of LPS-induced IL-8 gene expression in mononuclear phagocytes. Method : Human alveolar macrophages(HAM) and peripleral blood monocytes(PBMC) were isolated from healthy volunteers. Time and dose relationship of LPS-induced IL-8 mRNA expression was observed by Northern blot analysis. To evaluate the regulatory mechanism of LPS-induced IL-8 gene expression, pretreatment of actinomycin D(AD, $5{\mu}g/ml$) and cycloheximide(CHX, $5{\mu}g/ml$) was done and Northern blot analysis for IL-8 mRNA and ELISA for immunoreactive IL-8 protein in culture supernatant were performed. Results : 1) In HAM, dose and time dependent LPS-induced IL-8 mRNA expression was observed with peak mRNA level at 8 hours post-stimulation. 2) In PBMC, dose and time dependent LPS-induced IL-8 mRNA expression was also observed with peak mRNA level at 4 hours post-stimulation. 3) AD decreased expression of LPS-induced IL-8 gene expression at both mRNAand protein levels in both types of cells. 4) CHX decreased expression of LPS-induced IL-8 gene expression at protein level in both cell types but in HAM, superinduction of IL-8 mRNA was observed while decreased expression of IL-8 mRNA was observed in PBMC. Conclusion : Time and dose dependent LPS-induced IL-8 gene expression was observed in mononuclear phagocytes which is at least partly regulated pretranslationally. LPS-induced IL-8 mRNA expression in HAM needs no de novo protein synthesis and may be under the control of a labile repressor protein while de novo protein synthesis may be needed in PBMC.

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Effect of Conjugated Linoleic Acid on Nuclear Factor-${\kappa}B$ Activation and Tumor Necrosis Factor-${\alpha}$ Production in RAW 264.7 Cells Exposed to High Concentration of Glucose (고농도의 당에 노출된 RAW 264.7 세포에서 conjugated linoleic acid의 TNF-${\alpha}$ 생산과 NF-${\kappa}B$의 활성 효과)

  • Lee, Minji;Kang, Byeong-Teck;Kang, Ji-Houn;Yang, Mhan-Pyo
    • Journal of Veterinary Clinics
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    • v.29 no.5
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    • pp.361-367
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    • 2012
  • Diabetes-related complications in human and veterinary medicine have been shown to be associated with hyperglycemia-induced inflammation. It has been recently suggested that the onset of insulin resistance may be caused by over-production of inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$ from immune cells. Conjugated linoleic acid (CLA) regulates inflammatory response through modulation of TNF-${\alpha}$ expression. The objective of this study was to examine the effect of CLA on nuclear factor kappaB (NF-${\kappa}B$) p65 binding activity, inhibitory kappaB ($I{\kappa}B$)-${\alpha}$ expression, and TNF-${\alpha}$ production from high glucose-treated RAW 264.7 cells. CLA was added to RAW cells that had been previously cultured with low or high concentration of glucose. The levels of TNF-${\alpha}$ protein in the culture supernatant of RAW cells exposed to high concentrations of glucose were higher than those of cells exposed to low concentrations of glucose. The treatment with the high concentration of glucose in RAW cells increased levels of NF-${\kappa}B$ p65 binding activity and the decreased $I{\kappa}B-{\alpha}$ expression when compared with those of low glucose. The treatments in combination with CLA and glucose (low and high) glucose in RAW cells increased TNF-${\alpha}$ production when compared with that glucose alone. These treatments with CLA increased TNF-${\alpha}$ production in high glucose-treated RAW cells than those with low glucose. These treatments of CLA also showed higher NF-${\kappa}B$ p65 binding activity and lower $I{\kappa}B-{\alpha}$ expression in high glucose than those in low glucose condition. This suggests that CLA can increase NF-${\kappa}B$ p65 binding activity and TNF-${\alpha}$ production from high glucose-treated RAW 264.7 cells and is likely to promote hyperglycemia-induced inflammation.

Changes in the Blood Components Caused by Water Intake (물 섭취에 의한 혈액 성분 변화에 관한 연구)

  • Kim, Hyun-Kyung;Kim, Soo-Hwan;Ryu, Jae-Ki
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.3
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    • pp.227-232
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    • 2017
  • Although water is an essential component of the human body and is involved in many physiological processes, the effect of a steady and sufficient water intake on blood components has not well elucidated. Therefore, we investigated the changes in hematological parameters, high-sensitivity C-reactive protein (hs-CRP), and immunoglobulin G (IgG) after water intake in 13 healthy adults. They were divided into two groups: The control group (N=4), which consumed water ad libitum, and the experimental group (N=9), which consumed 2 L of water per day. Two weeks later, blood cell counts, hematocrit, and hemoglobin content had increased in the experimental group, although not significantly (p>0.05); however, there was a significant increase in the mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) (p<0.05; and p<0.01, respectively), and a significant decrease in the mean platelet volume (MPV) (p<0.05). Of the immunologic parameters, a non-significant decrease in the concentration of hs-CRP, an indicator of cardiovascular disease risk, was observed (p>0.05). However, there was a dramatic and significant increase in the concentration of IgG (p<0.05). In conclusion, a steady and sufficient water intake may contribute to alleviate anemia by increasing hemoglobin. Additionally, it may decrease the risk of cardiovascular disease by decreasing platelet activation and concentration of hs-CRP. Furthermore, a steady intake of water may improve immune function by increasing the concentration of the components of humoral immunity.