• 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.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.11 no.1
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• pp.69-81
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• 1998

The aim of this study was to investigate the effects of Chungdaesan(CDS) by various administration routes on skin anaphylactic reaction. The most classic and popular skin reaction in vivo is passive cutaneous anaphylaxis(PCA) In this study, therefore, the author investigated the effect of CDS on PCA reaction activated by anti-dinitrophenyl immunoglobulin E antibody The results showed that CDS potently suppressed orally, topically, intraperitoneally, and intradermally administered. However, it did not show suppressive activity when intravenously administered. In addition CDS significantly inhibited anti-DNP IgE induced mast degranulation in mice skin. Moreover, CDS suppressed anaphylactic histamine release from mast cells induced by anti-dinitrophenyl immunoglobulin E antibody. These results indicate that CDS suppresses the PCA reaction by stabilization of mast cells in vivo and in vitro am] also suggest that the differential activity following administration routes may be caused by the difference of bioavailability.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.6
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• pp.1487-1492
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• 2003

Gunjung-tang(GJT) has been reported at Shanghanlun(傷寒論), which has been used for the treatment of general weakness, digestive organ disease and the treatment of allergic disorders in clinical medicine. However, its effect in experimental models remains unknown. In a rat and mouse model, the role of GJT was examined in mast cell-dependent allergic reactions and secretion of inflammatory cytokines. GJT inhibited anti-dinitrophenyl IgE antibody-induced passive cutaneous anaphylaxis reaction by Intraperitoneal pretreatment. GJT inhibited both IL-1β and TNF-α secretion on egg albumin induced allergic tissues and compound 48/80 induced thymus. Furthermore, GJT enhanced Band T lymphocytes proliferation. Our findings provide evidence that GJT inhibits the IgE-dependent allergic reactions and inflammatory cytokines secretion, and enhanced immune response.

• Parasites, Hosts and Diseases
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• v.31 no.3
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• pp.231-238
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• 1993

Effects of Paragonims westermai infection were observed in mice on the change of serum IgE level, the number of peripheral eosinophils and the distribution of Thy $1.2^{+}{\;}(CD3),{\;}L3T4^{+}{\;}(CD4),{\;}and{\;}Lyt-2^{+}$ (CD8) splenic T Iymphocytes without mitogen serum IgE increased at 3 weeks after the infection and reached a peak on week 4 and maintained high levels of IgE until the 23r6 week. Peripheral eosinophil numbers Increased at the second week and attained peak level on week 9. The frequency of $L3T4^{+}$(CD4) and $Lyt-2^{+}$ (CD8) T Iymphocytes decreased slightly until 4 weeks after the infection, but not significantly. Absolute number of $L3T4^{+}{\;}and{\;}Lyt-2^{+}$ T Iymphocytes, and the ratio of L3T4/Lyt-2 were not markedly changed over the period of observation. The frequency of Thy $1.2^{+}$ (CD3) T lymphocytes in the infected group slightly decreased until 4 weeks after the infection and showed significant reductions at the 2nd and 4th week of the infection (p < 0.05).

• Proceedings of the Korean Society of Applied Pharmacology
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• 1997.04a
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• pp.95-95
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• 1997

Among the various receptor molecules discovered so far the ${\beta}$2-adrenergic receptors have been regarded as excellent model systems for the so called 7 transmembrane helix receptor and have been the focus of extensive studies. For the analysis of receptor structure and function a monoclonal antibody plays a crucial role, thus providing useful tools for the study of receptor. However, because of the minute quantity of receptor molecules which could be obtained from natural sources, the generation of specific monoclonal antibody against receptor molecules from the purified receptors has been regarded as virtually impractical in consideration of cost and experimental times. The purpose of the present study was to generate and characterize a monoclonal antibody against human ${\beta}$2-adrenergic receptor. For the production of antibody, C-terminal regions of the human ${\beta}$2-adrenergic receptor was produced as a fusion protein with Glutathion S-transferase (GST) in E. Coli. The expression of the fusion protein was identified by SDS-PAGE and Western blot using monoclonal anti-GST antibody. The fusion protein was purified to an apparent homogeniety by affinity chromatography with Glutathion Sepharose CL-4B and used as an antigen for the immunization of BALB/C mice. The Production of monoclonal antibody was achieved by fusion of the immunized spleen cells and SP/2-0 myeloma cells. Positive hybridomas were screened by ELISA and were cloned by two consecutive rounds of limiting dilution. The monoclonal antibody produced in this study (mAb${\beta}$C02) was IgM type and purified by immunoaffinity chromatography using anti-mouse IgM agarose as an affinity matrix. MAb${\beta}$C02 showed strong and specific immunoreactivity against both the fusion protein and human ${\beta}$2-adrenergic receptor in ELISA and Western blot. The molecular weight of immunoreactive band was 64 kDa and exactly coincided with the previously reported molecular weight of ${\beta}$2-adrenergic recepters. The results of the present study suggest that mAb${\beta}$C02 may be used for the study of receptor function and regulation in normal or nonphysiological status.

• Annals of Clinical Neurophysiology
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• v.15 no.1
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• pp.27-29
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• 2013

• The Journal of Korean Obstetrics and Gynecology
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• v.23 no.4
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• pp.1-9
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• 2010

Purpose: This study was designed to investigate enhancing the immunogenicity effects of Platycodon grandiflorum(PG) on adaptive immune system. Methods: To investigate the effect of PG as an adjuvant, we used the ovalbumin (OVA) as an antigen at first. The proliferation of lymphocytes, the antibody titer, the subisotypes of antibodies and the production of cytokines were measured. Results: The proliferation of lymphocytes and the antibody titer were increased after PG treatment. The increased subisotypes of antibodies were IgG2 and IgG3 induced from T1-helper cells. However IgE induced from T2-helper cells was decreased. The production of cytokines derived from T1-helper cells was increased but that from T2-helper cells was decreased. Conclusion: It is supposed that PG has an immunogenicity effect as an adjuvant on adaptive immune system.

• The Journal of the Korean Society for Microbiology
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• v.22 no.3
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• pp.197-208
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• 1987

Chlamydia trachomatis has now shown that this interesting intracellular parasite is a cause of nongonococcal urethritis, infantile pneumonia, pelvic inflammatory disease and epididymitis, in addition to lymphogranuloma venerum and inclusion conjunctivitis. There are several diagnostic methods for C. trachomatis, but the method using monoclonal antibody is the most sensitive and specific. The hybride cell were prepared by fusion of myeloma cell($P_3X_{63}\;Ag_8{\cdot}V_{653}$) of mouse and lymphocyte of mouse(BALB/c) that were immunized with formalin killed C. trachomatis serotype D. The cell mixtures after fusion were dispensed into 640 wells of the 96 well culture plates and continuously cultured in HAT medium for 2 weeks. The supernatants of culture media in 83(13%) wells were reacted with C. trachomatis, which were determined by enzyme-linked immunosorbent assay in 96 well microplate. The clones that secreted antibody to C. trachomatis were cloned by limiting dilution. Only six monoclones secreted antibody to C. trachomatis. The antibody titer of ascitic fluid that collected from same BALB/c mice bearing hybridoma cells was above 1:100,000. These monoclonal antibodies that were IgG reacted with elementary and reticulate bodies of all serotypes(Ba, D, E, F, G, H, J and LGV type-I) using ELISA and indirect immunofluorescence stain, but there were no cross reaction with other bacteria(coagulase negative Staphylococcus, Proteus and E. coli). We concluded these six monoclones secreted the same monoclonal antibody to C. trachomatis. The sensitivity and specificity of the monoclonal antibody compared with Microtrak(confirmatory test of C. trachomatis, Syva) was 100%, respectively.

• YAKHAK HOEJI
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• v.49 no.5
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• pp.386-391
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• 2005

The discovery of drugs for the treatment of mast cell-mediated allergic disease is a very important subject in human health. The Amomum xanthiodes (Zingiberaceae) has been used for centuries as a traditional medicine in Korea and is known to have an anti-inflammatory effect. However, its specific mechanism of action is still unknown. In this report, we investigated the effect of hot water extract from Amomum xanthiodes (EAX) on the mast cell-mediated allergic reaction and studied its possible mechanisms of action. EAX inhibited compound 48/80-induced systemic anaphylaxis and serum his­tamine release in mice. EAX decreased the passive cutaneous anaphylaxis reaction activated by anti-dinitrophenyl (DNP) IgE antibody. EAX dose-dependently reduced histamine release from rat peritoneal mast cells activated by compound 48/80 or anti-DNP IgE. EAX increased cAMP and decreased compound 48/80-induced intracellular $Ca^{2+}$ levels. Our findings provide evidence that EAX inhibits mast cell-derived allergic reactions, and also demonstrate the involvement of cAMP and intracellular $Ca^{2+}$ in these effects.

• Pediatric Infection and Vaccine
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• v.4 no.1
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• pp.126-132
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• 1997

Measles outbreak in the world was decreased since measles vaccine had been introduced. Although vaccination rate is high, measles was not eradicated and measles reappeared among vaccinated children. We measured measles-specific antibody from the vaccinated and unvaccinated groups who had experienced apparent measles in the Seongnam city in 1993. The results were as follows. 1) The data included total 126 children (M:F=1 : 1). Age distribution of measles outbreak revealed 6 children in 5yr, 11 in 6yr, 20 in 7yr, 39 in 8yr, 22 in 9yr, 11 in 10yr, 11 in 11yr, and 6 in 12yr. 2) MMR vaccination rate was 78.6%(99/126) in the children who had experienced measles. Positive rate of measles-specific IgM Ab was 80.8% (80/99) among the vaccinated group and among 9E.6.% (25/27) the unvaccinated. 3) Positive rate of measles-specific IgG Ab was 90.9% (90/99) among MMR-vaccinated group, and 85.2% (23/27) in unvaccinated group. In conclusion, measles-specific IgM antibody have been detected more than 1 month in most patients. The relatively high proportion of measles-specific IgM positivity may mean primary vaccine failure. To booster the antibody titers and to prevent measles epidemic in school-aged children, revaccination of measles should be considered.