• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.18 no.3
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• pp.18-25
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• 2005

Objectives: It has a growing interest in the prevention and medical treatment of allergic rhinitis. According to many studies, it's known that Sinicheongpye-um has the inhibitory effect on the allergic rhinitis. We have studied effect of the mice on OVA-induced Production of IL-4, IL-5, $Interferone-{\gamma}$ by Murine Splenocytes and effect of OVA-induced Total IgE. Methods: Mixing Ovalbumin(OVA) $10{\mu}g$ into PBS(phoshate buffered saline) and $Al(OH)_3$, gel solution and changing into $1\;m{\ell}$, we made it into OVA solution. That was administered to normal group. After the last administration into abdominal cavity, we caused allergic rhinitis in nasal cavity of mouse of control group and sample group administering 0.1% solution dropwise 3 times a day for 7 days. Keeping separated serum at -20 degree and after refloating spleen cells, cultivating the cells and centrifuging the upper liquid and keeping it at -20 degree, we measured the amount of IL-4. IL-5, $Interferone-{\gamma}$ and OVA-induced Total IgE by ELISA. Results: 1. In Total IgE, Sinichengpae-um treated group was proved significant inhibitory effect.(p<0.05) 2. In IL-4 study, Sinichengpae-um treated group was proved significant inhibitory effect.(p<0.005) 3. In IL-4 study, Sinichengpae-um treated group was proved significant inhibitory effect.(p<0.001) 4. In $Interferone-{\gamma}$ study, Sinichengpae-um treated group showed a increasing tendency. Conclusion: Based on the above result, it is considered that Sinichengpae-um has the inhibitory effect on the allergic rhinitis of mice and suggested thai it could be used in relieving patients of the symptoms which are caused by allergic rhinitis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.5
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• pp.547-552
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• 2017

The viral hemorrhagic septicemia virus (VHSV) has an extensive host range, and infects farmed and wild fish inhabiting both freshwater and marine ecosystems. Enzyme-linked immunosorbent assay (ELISA) is highly useful in diagnosing viral hemorrhagic septicemia. However, ELISA shows high, non-specific background reaction with fish antibodies. In this study, we optimized the antigen and antibody concentrations used for detecting specific antibodies in VHSV-infected olive flounder to reduce non-specific binding, and improve the sensitivity of ELISA. The results suggested that OD (optical Density) values were valid when ELISA was performed with $0.1{\mu}g/well$ of virus, involving blocking with blocking buffer (Roth, Roti-Block), 1:300-1:600 dilution with flounder antisera, and 1:1000 dilution with anti-flounder IgM and HRP-conjugated goat anti-mouse IgG for detecting the VHSV antibody in flounder sera. Furthermore, 11 different VHSV strains isolated in Korea from 2012 to 2016 were used to infect the fish. The results showed no correlation between viral pathogenicity and antibody production. This research is a basic study on the application of antibody detection in the diagnosis of viral hemorrhagic septicemia in the olive flounder.

• Journal of Acupuncture Research
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• v.33 no.2
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• pp.21-34
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• 2016

Objectives : The aim of the present study is to examine the effect and mechanism of Erycibae Caulis and Corydalis Tuber Pharmacopuncture (ECP) on a mouse model with collagen induced rheumatoid arthritis (CIA). Methods : We evaluated the Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Creatinine, and the Blood urea nitrogen (BUN) of serum to examine the safety of this study. In vivo, we compared the results of the non-treated group, the normal saline pharmacopuncture treated control group, the indomethacin treated group and the ECP group. We evaluated rheumatoid arthritis manifestation and the Rheumatoid Arthritis Index (AI). Also, immune cells in blood affected by ECP were evaluated by calculating the level of white blood cells (WBC), neutrophil, lympocytes and monocytes. Next, the level of Immunoglobulin M (IgM), Immunoglobulin G (IgG), Interleukin (IL)-$1{\beta}$, IL-6, IL-17, Tumor Necrosis Factor (TNF)-${\alpha}$ and Granulocyte-macrophage Stimulating Factor (GM-CSF)in serum were measured. We examined the imaging of cartilage degeneration using micro CT-arthrography of the hind paw. Additionally, we examined the effects of reducing bone volume (BV) ratio and bone surface/bone volume (BS/BV) ratio with 3D Micro-CT. Finally, we did a histopathologic examination analysis. Results : The absence of liver and kidney toxicity was evident. In vivo, edema of the joints of the ECP group decreased greatly in macroscopic observation. AI measurement of the ECP group also decreased significantly compared to the control group. The level of WBC, neutrophil, lympocytes, and monocytes in the blood decreased but there was no statistical significance of this data. IgM of the ECP group decreased significantly compared to the control group. IL-$1{\beta}$, IL-6, TNF-${\alpha}$, and GM-CSF production of the ECP group decreased significantly compared to the control group. As a result of examining joint condition with 3D micro CT, deformation and destruction of the joint was shown to have decreased. Bone density of ECP group increased at a statistically significant level compared to the control group. Degree of joint inflammation of ECP group decreased significantly compared to the control group. After H&E and M-T staining, infiltration of immune cells, subsidence of the cartilage, damage to the synovial cells and joint erosion decreased. Conclusion : This study showed that ECP hindered the process of rheumatoid arthritis and protected joints and cartilage.

• Archives of Pharmacal Research
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• v.24 no.6
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• pp.557-563
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• 2001

Oral tolerance is thought to play a role in preventing allergic responses and immune-mediated diseases. An improved mouse model of the oral tolerance to Japanese cedar pollen (JCP) as antigen was developed in order to detect induction of the tolerance, and the immunological characteristics of this model were also elucidated. Oral tolerance was induced by C3H/ HeN mice given an oral administration of 10 mg JCP 7 days before immunization with an i.p. injection of 0.1 mg JCP in complete Freunds adjuvant (CFA). The effects of oral JCP on systemic immunity were assessed by enzyme-linked immunosorbent assay (ELISA) of immunoglobulin (Ig) levels in serum collected on day 7 or 14 after immunization. Oral tolerance to JCP was adequately induced on day 7 after immunization and was more effective in C3H/HeN mice than in BALB/c mice. The tolerance was primarily concerned with the decreased serum levels of antigen-specific IgG. In these mice, oral administration of JCP also suppressed various immune responses to the antigen including delayed-type hypersensitivity (DTH), total Igl level and anti-JCP IgGl level. The suppression of these immune responses by the oral antigen was associated with a significant reduction in interleukin-4 (IL-4) production. These findings therefore indicate that this C3H/HeN mice model has potential use in detecting the induction of oral tolerance by JCP and suggest that this tolerance model may be effective in the treatment and prevention of allergic responses caused by the antigen.

• Korean Journal of Veterinary Research
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• v.35 no.4
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• pp.769-776
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• 1995

This study was undertaken to develop the monoclonal antibody(MAb) for lymphocytes of Korean native cattle by the cell hybridization of myeloma P3/NS-1/ 1-Ag-4-1 and spleen cells from BALB/c mice hyperimmunized with nylon wool column eluted peripheral T lymphocytes of Korean native cattle. The isotype of MAb KCT-14 against T lymphocyte was mouse $IgG_1$. KCT-14 positivity of mononuclear cells(MNC) from peripheral blood lymphocytes, nylon wool nonadherent and adherent-lymphocytes was 41.7%, 58.4% and 22.6%, respectively. And that of mesenteric lymph node-, spleen and thymus-MNC was 43.3%, 40.2% and 33.6%, respectively. Immunoperoxidase staining of frozen tissue sections showed that the MAb positive cells were located in the medulla of the thymus and in the paracortical area and the mantle zone of the germinal center in the lymph nodes. These results indicated that KCT-14 was one of the MAb for investigate of T lymphocyte subpopulations in the Korean native cattle.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.51 no.3
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• pp.328-331
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• 2018

We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against nervous necrosis virus (NNV, RGNNV genotype). We established six hybridoma clones secreting MAbs against NNV antigen: 2B1, 2B11, 2C12, 13C1-1, 13C1-2 and 14D11. All six MAbs belonged to the IgG2a isotype with a kappa light chain and their reactivity recognized against the 41 kDa coat protein of NNV by Western blot analysis. The affinity constants of the six MAbs were measured by enzyme-linked immunosorbent assay (ELISA). All six MAbs reacted with two NNV isolates (SgNag05 and Gemunodo06), while no reactivity was observed with five know fish viruses, namely marine birnavirus, infectious pancreatic necrosis virus, viral hemorrhagic septicemia virus, hirame rhabdovirus, and infectious hematopoietic necrosis virus. Moreover, high ELISA optical density (OD) values (0.87-1.42) were observed in the brain tissues of NNV-infected sevenband grouper, while low OD values (less than 0.12) were recorded in the brain tissues of uninfected fish. These results suggest that these six MAbs are highly competent and useful for the detection of NNV with the RGNNV genotype.

• Journal of Environmental Health Sciences
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• v.21 no.3
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• pp.16-22
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• 1995

This study was designed to investigate the antibody production to sheep red blood cells(SRBC) and proliferation of mitogen-stimulated spleen cells in Balb/c mice which received cadmium chloride. The mice were divided into three independent groups which were one control and two experimental groups by the cadmium treatment or not. No specific treatment was done for the control group. One of two experimental groups, which is called 'pre-treatment group' in this paper, was subcutaneously injected with low dose of cadmium chloride(0.5 mg/kg/day) for 5 consecutive days before the primary SRBC immunization. The other called 'non-pretreatment group' was only pretreated with normal saline. Both experimental groups were intraperitoneally injected with high dose of cadmium chloride(5 mg/kg) 8 hours before the primary immunization. Mice were intraperitoneally immunized twice with 2% SRBC suspension containing $10^8$ cells. The results obtained were as follows, 1. The PFG responses to SRBC were significantly increased in two experimental groups, cadmium pretreatment and non-pretreatment compared with that of control group(p<0.05). 2. The total antibody titers to SRBC in cadmium treated groups were similar to that of control group, but titers of IgG antibody were significantly elevated(p<0.01). 3. The proliferation response of spleen lymphocytes to various mitogens was suppressed in proportion to the concentration of cadmium and the degree of cadmium accumulation in liver was increased in the cadmium treated groups. These results suggest that cadmium chloride could affect on mouse immune response, especially its cell mediated immune response could be decreased while its humoral immune response could be increased, which may not be influenced by the administration methods or pretreatment of cadmium to mouse.

• 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.

• 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.

• The Journal of the Korean Society for Microbiology
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• v.22 no.4
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• pp.377-392
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• 1987

Monoclonal antibody to the Escherichia coli heat-stable enterotoxin(ST) was produced to develop a rapid and convenient diagnostic method to the ST. The toxin was purified from culture supernatant of enterotoxigenic E. coli O148H28($ST^+/LT^+$) and conjugated to bovine serum albumin(BSA). The ST-BSA conjugate was used to immunize BALB/c mice and the immune spleen cells from these mice were fused with $P3{\times}63$ Ag8.V653 plasmacytoma cells. Hybridomas were screened by ELISA and positive hybridomas were cloned by limiting dilution. Finally, one stable clone (AS36) specific to ST was selected for further growth and characterization. Antibody titers of culture supernatant and ascitic fluid from BALB/c mice were 1:1,024 and 1:20,480 respectively in ELISA. The isotype and subclass of monoclonal antibody was IgG1 in sandwich ELISA. To test the neutralizing effect of monoclonal antibody on toxin activity of ST, mixture of ascitic fluid and ST was assayed by infant mouse assay and this monoclonel antibody was proved to be a neutralizing antibody. The titer of ascitic fluid which completely neutralized biological activity of 4 units of ST was 1:4. Purified ST was quantitatively measured by competitive ELISA and minimum amount of ST detectable by this assay was 250pg, which was an amount six-fold smaller than that detectable by infant mouse assay. Four reference strains of enterotoxigenic E. coli from WHO were detected by competitive ELISA and highly specific, sensitive and reproducible result was obtained.