• The Journal of the Korean Society for Microbiology
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• v.21 no.1
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• pp.151-161
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• 1986

In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

• Food Science of Animal Resources
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• v.34 no.5
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• pp.570-575
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• 2014

This study was conducted to compare the effects of two forms of radiation (electron and X-ray; generated by an electron beam accelerator) on the conformation and antigenic properties of hen's egg albumin, ovalbumin (OVA), which was used as a model protein. OVA solutions (2.0 mg/mL) were individually irradiated by electron beam or X-ray at the absorbed doses of 0 (control), 2, 4, 6, 8, and 10 kGy. No differences between the two forms of radiation on the structural properties of OVA were shown by spectrometric and electrophoretic analyses. The turbidity of OVA solution increased and the main OVA bands on polyacrylamide gels disappeared after irradiation, regardless of the radiation source. In competitive indirect enzyme-linked immunosorbent assay, OVA samples irradiated by electron beam or X-ray showed different immunological responses in reactions with monoclonal and polyclonal antibodies (immunoglobulin G) produced against non-irradiated OVA. The results indicate that electron beam irradiation and X-ray irradiation produced different patterns of structural changes to the OVA molecule.

• Korean Journal of Clinical Laboratory Science
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• v.41 no.1
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• pp.42-46
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• 2009

Unlike most bacteria, Treponema pallidum subspecies cannot be readily isolated or sustained in cell culture for numerous generations. In korea, two non treponemal tests are currently considered as standard; the VDRL slide test and RPR card test. These tests are based on an antigen composed of an alcoholic solution containing measured amount of cardiolipin, cholesterol, and sufficient purified lecithin to produce reactivity. The nontreponemal reagin tests measure immunoglobulin M (IgM) and IgG to lipoidal material released from damaged host cells as well as to lipoprotein-like material and possibly by cardiolipin released from the treponemes. The object of the evaluation was to evaluate the performance of the Mediace RPR kit on the automated biochemistry analyzer system as a method for screen method of syphilis as well as to identify BFP possibility. For evaluation of routine screening test, a total 2,380 specimens tested by Mediace RPR from 28th Oct, 2007 to 22th Feb, 2008. For evaluation of BFP possiblility, we measured samples which have potential BFP reaction in Syphilis test such as ANA (anti-nuclear antibody) positive (135 samples), CRP (C-reactive protein) positive (100 samples), RF (Rheumatoid factor) positive (26 samples), and other potential BFP cases (17 samples) including total 278 samples. These samples were tested quantitative test Mediace RPR with Hitachi 7600 P module. For comparison with current manual test, VDRL slide test were performed. Of these 2380 specimens, 2350 were negative, 30 were positive, and one were positive with TPHA. Both methods agreed for 2356 (98.9%) samples. Of the 30 samples showed positive results over 1.0 R.U, 6 samples showed positive results with VDRL test. Of these 6 samples, 1 samples showed positive with TPHA test. The combination of the Automated Biochemistry analyzer and VDRL test for retest can be increase efficiency of syphilis screening test.

• The Korea Journal of Herbology
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• v.29 no.6
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• pp.95-102
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• 2014

Objectives : The purpose of this study is to prove the effect of GamiBangkeehwangkee-tang (f$\acute{a}$ngj$\check{i}$hu$\acute{a}$ngq$\acute{i}$-t$\bar{a}$ng, BHT) ethanol extract on the immunity and rheumatoid arthritis related inflammatory cytokines. Methods : We checked viability in RAW 264.7 cell after treat by BHT. Then we measured inflammatory and immunity factors of DBA/1 mice with rheumatoid arthritis induced by collagen after BHT oral administration. Also, we checked micro-CT image, bone volume and bone inflammation ratio in micro-CT and structural parameter test. Results : BHT showed cell viability of 95% or higher in all concentration in RAW 264.7 cells. BHT treated group decreased level in serum of IgM and IgG test by 36% and 25% respectively. And BHT treated group showed significant decrease in WBC, neutrophil, lymphocyte and monocytes immune cell ratio in blood by 47%, 22%, 56% and 85% respectively. Also, BHT treated group decreased level in serum of $IL-1{\beta}$, IL-6, IL-17, $TNF-{\alpha}$ and hs-CRP tests by 29%, 33%, 32%, 24% and 56% respectively. Finally, BHT treated group showed increase ratio of bone volume that decrease ratio of bone inflammation. Conclusions : In this study, the results were observed rheumatoid arthritis factors cytokine decrease in serum. And BHT showed immunoglobulin and immune cells ratio decrease in serum and blood. Also, BHT depending on effects of inflammatory and immunity in hs-CRP test, micro-CT, structural parameter test. Thus, these results can used as a effective drug of BHT for inflammation and immunity.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.11
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• pp.4459-4463
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• 2014

Background: Gastric cancer is the second leading cause of cancer-related deaths worldwide and infection with H. pylori is considered essential for its development. Helicobacter pylori infects more than 50% of the world's population with higher prevalence in developing countries than developed countries. The prevalence of H. pylori varies in different societies and geographical locations. The objectives of this study were to estimate the seroprevalence and determine the risk factors of H. pylori infection in dyspeptic patents in Ethiopia. Materials and Methods: A cross-sectional study involving 209 dyspeptic patients was carried out from February 15 to April 30, 2013. Five to ten ml venous blood was collected from each dyspeptic patient and analyzed for detection of Helicobacter pylori immunoglobulin (IgG). The socio-demographic characteristic, hygienic practices, alcohol consumption, sources of drinking water and types of latrine were also obtained with a pre-tested questionnaire. Results: The overall seroprevalence of Helicobacter pylori was 72.2%. There was statistically significant difference in the prevalence of H. pylori among age groups (p=0.02). Seroprevalence of H. pylori was higher in those patients who used unprotected surface water (76.4%) than those with access to piped tap water (65.9%). There was also statistically significant differences in prevalence of H. pylori with the habit of hand washing before meal (p=0.01) and alcohol consumption (p=0.001). Conclusions: The prevalence of H. pylori was high in the study area and increased with age of dyspeptic patients. Alcohol consumption and the type of drinking water are risk factors that have associations with the prevalence of H. pylori. Molecular epidemiological techniques can show a true picture of H. pylori and improvement in the drinking water quality is recommended.

• Journal of Veterinary Clinics
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• v.8 no.1
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• pp.1-10
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• 1991

The B-lymphocyte differentiation from committed B-cell progenitors to antibody-secreting cells was discussed. B-cell progenitors derived from hematopoietic stem cells undergo the rearrangement of immunoglobulin(Ig) gene. The earliest cells as B-cell precursors have cytoplasmic Is(${\mu}$ chain). The entire Is molecule is expressed on the surface after synthesis of L chain. The resting B cells(Go stage) stimulated by binding antigen via Ig-receptors are activated(G$_1$ stage) and followed by proliferation(S stage), coupled with further selection(affinity maturation. class switch). The production of antibody against a particular antigen depends on the activation of B cells with surface Is capable of reacting with that antigen. This process does not occur in isolation but is controlled by helper and suppressor T cells and antigen presenting cells(APC). The mechanism of T cell-dependent B-cell response for production of antibody is largely explained by the cell to cell cooperation and soluble helper factors of T cells. 1) The antigen specific B cells and helper T cells are linked by Is-receptors, leading to the delivery of helper signals to the B cells. 2) Helper T cells recognize the processed antigen-derived peptides with the MHC class II molecules(la antigen) and is stimulated to secrete B-cell proliferation and differentiation factors which activate B cells of different antigenic specificity. The two models are shown currently 1) At low antigen concentration, only the antigen-specific B cell binds antigen and presents antigen-derived peptides with la molecules to helper T cells, which are stimulated to secrete cytokines(IL-4, IL-5, etc.) and 2) At high antigen concentration, antigen-derived peptides are presented by specific B cells, by B cells that endocytose the antigens, as well as by APC Cytokines secreted from helper T cells also lead to the activation of B cells and even bystander B cells in the on- vironmment and differentiate them into antibody-secreting plasma cells.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.7
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• pp.889-897
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• 2011

Mycoplasma Pneumonia of swine (MPS) decreases the daily growth of pigs, and, co-infection with a virus sometimes causes severe pneumonia. Genetic selection of pigs resistant to the pulmonary MPS lesion might solve the economic loss due to MPS in animal production. Here, we examined the immunophenotype of Landrace line (Miyagino L2), genetically selected to reduce the incidence of pulmonary MPS lesion for 5 generations in Miyagi Prefecture Animal Industry Experiment Station. Although this line is expected to be resistant to the pulmonary MPS lesion, the biological characteristics of its immune function are not clear. We investigated details of the immunorelated phenotype of Miyagino L2 at the hematological and molecular biological level, including cytokine expression, and compared the results with that of non-genetically selected Landrace. Miyagino L2 showed decreased antigen-specific IgG and IgM production and increased CD8-positive T-cell population, and high levels of cortisol concentration, suggesting that the MPS-resistant phenotype is associated these immunological differences. Additionally, T-cell CD4 expression was highly correlated with the MPS expected breeding value. Although the detailed mechanisms underlying this high correlation remain unknown, our result suggested that the genetic selection of the expression level of CD4 might be useful to improve MPS resistance in pig production.

• Childhood Kidney Diseases
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• v.19 no.2
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• pp.176-179
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• 2015

Rituximab (RTX), a monoclonal antibody against the B-cell marker CD20, is commonly used as a treatment for antibody-mediated diseases or B-lymphocyte-mediated diseases. Destruction of B cells may reverse the disease course in many conditions; however, patients who are treated with RTX cannot respond appropriately to de novo infection due to lack of B lymphocytes. Here, we report one such case. A 7-year-old renal allograft recipient presented with severe anemia due to parvovirus infection after RTX treatment. The patient had focal segmental glomerulosclerosis and had received cadaveric kidney transplantation 6 months previously. She was treated with high-dose steroid for acute rejection and RTX for Epstein Barr Virus infection 3 months previously. At presentation, her hemoglobin level was 5.4 g/dL and leukocyte and platelet counts were normal. She had microcytic normochromic anemia and high viral load of parvovirus B19(70,578 copies/mL). Intravenous immunoglobulin ($200mg/kg{\cdot}d$) treatment controlled the progression of anemia and parvovirus infection. De novo parvovirus infection during the B lymphocyte-depletion period may have precipitated the severe anemia in this case. Close monitoring of infection is required after RTX therapy.

• Korean Journal of Veterinary Research
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• v.53 no.4
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• pp.217-224
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• 2013

Transgenic plants have been tested as an alternative host for the production and delivery of experimental oral vaccines. Here, we developed transgenic potatoes that express the major antigenic sites A and D of the glycoprotein S from transmissible gastroenteritis coronavirus (TGEV-$S_{0.7}$) under three expression vector systems. The DNA integration and mRNA expression level of the TGEV-$S_{0.7}$ gene were confirmed in transgenic plants by PCR and northern blot analysis. Antigen protein expression in transgenic potato was determined by western blot analysis. Enzyme-linked immunosorbent assay results revealed that based on a dilution series of Escherichia coli-derived antigen, the transgenic line P-2 had TGEV-$S_{0.7}$ protein at levels that were 0.015% of total soluble proteins. We then examined the immunogenicity of potato-derived TGEV-$S_{0.7}$ antigen in mice. Compared with the wild-type potato treated group and synthetic antigen treated group, mice treated with the potato-derived antigen showed significantly higher levels of immunoglobulin (Ig) G and IgA responses.

• Journal of Nutrition and Health
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• v.27 no.5
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• pp.483-494
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• 1994

The purpose of this study was to investigate the effect of nutritional status on the cell-mediated and humoral immunity in female college students. The nutritional status of twenty subjects was determined by six-days food records, anthropometric measurements, and biochemical assessments of serum nutrients. Cell-mediated and humoral immunity of the subjects was analyzed by in vivo and in vitro assessments. The results were summerized as follows : First, The average daily energy intake was 1437Kcal(CHO : PRO : FAT = 61:13:26), which corresponds to 71.9% of RDA. Anthropometric measurements showed that 50% of the subjects was under-weight(BMI<20), only 5% was over-weight(25