• The Korean Journal of Pain
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• v.18 no.1
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• pp.52-55
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• 2005

Complications following a well conducted epidural steroid injection are rare. A 50-year-old man developed a headache and neck stiffness 2 days after a lumbar epidural steroid injection. Under the impression of aseptic meningitis, fluid and nonsteroidal anti-inflammatory drug therapy was started immediately after cerebrospinal fluid (CSF) sampling. The CSF was turbid, and revealed a white blood cell count, protein, glucose and pressure of $550/{\mu}l$ (98% lymphocyte), 107.9 mg/dl, 48 mg/dl (serum 113 mg/dl) and $17cmH_2O$, respectively. The CSF stain and culture, and antibody test and polymerase chain reaction for pathogens were negative. A computed tomography (CT) scan of the brain revealed no abnormality, and a chest roentgenogram and the results of the neurological examination were normal. Under the impression of aseptic meningitis, the condition was managed conservatively, without antibiotics. Seven days later, the clinical symptoms had improved, and the patient discharged.

• Korean Journal of Pharmacognosy
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• v.32 no.3 s.126
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• pp.226-232
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• 2001

To explore the possible cancer therapeutic application of "Bo-yang-hwan-oh-tang" (BH), a herbal medicinal recipe used for improvement of blood stasis, we have examined its direct cytotoxicity against tumor cell, and induction of cytotoxic activity of lymphocytes. Water extract of BH alone did not exhibit direct cytotoxicity to Yac-1 target cells even with high concentrations (10 mg/ml). By exposure for 3 days, BH did not induce any nonspecific cytotoxic activity of mouse spleen cells, either, when assessed in a 4 hr $^{51}Cr-release$ assay. However, when BH was added during CD3 stimulation of non-adherent spleen cells, non-specific CTL activity was markedly promoted in a dose dependent manner. In contrast, BH did not alter activated NK cell activity following IL-2 stimulation. These data suggest that BH does not induce but upregulates non-specific CTL effecter function and that activated NK cell does not respond to BH. For elucidation of the mechanism underlying this function of BH, time kinetic study for IL-2 production using ELISA was undertaken. IL-2 production following CD3 stimulation was significantly augmented and higher level of IL-2 is sustained over 3 days in the culture medium by BH treatment. Moreover, addition of exogenous IL-2 during CD3 stimulation resulted in a similar level of cytotoxicity between control and BH-treated culture. These data indicate that the BH-mediated upregulation of non-specific CTL activity is contributed by augmentation of IL-2 production. Our data imply the possible application of BH for combination therapy of cancer with non-specific activator.

• The Journal of the Korean Society for Microbiology
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• v.18 no.1
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• pp.59-65
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• 1983

To investigate the effects of placental serum to in vitro culture of the normal human lymphocytes the peripheral blood lymphocytes were isolated by ficoll-hypaque separation method in 20 healthy human adults. The blast transformation respone of lymphocytes to mitogens were observed by stimulation with PHA($25{\mu}g/ml$) and Con A($25{\mu}g/ml$) using RPMI 1640 media containing 20% placental serum(PS), tetal calf serum(FCS), or humans AB serum(AB). And one-way mixed lymphocyte culture test was performed between these unrelated person compounded into stimulators and responders to investigate the effect of placental serum. The following results were obtained. 1. In 20 experimental cases, these were no significant diffenence between FCS, AB, and PS in untreated control cultures. 2. In PHA-treated cultures, whereas the blast transformation rate of the FCS groups and AB groups were $40.8{\pm}4.3%$ and $44.6{\pm}4.3%$ respectively, that of PS groups were $21.7{\pm}3.4%$, Similar results were obtained in Con A-meated cultures. Therefore, placental serum inhibited the mitogenic response of lymphocytes significantly. 3. In MLC tests, stimulation index of the FCS groups and AB groups were $18.5{\pm}3.5$ and $20.1{\pm}3.3$ respectively. But placental serum inhibited MLC response of lymphocytes significantly($7.4{\pm}1.9$).

• KSBB Journal
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• v.26 no.5
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• pp.386-392
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• 2011

Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig), an immunosuppressive agent, was expressed in transgenic rice cells using RAmy3D promoter and RAmy1A signal peptide for the inducible production and secretion into culture media by sugar depletion. In this study, sodium butyrate was used as a small molecular enhancer (SME) to enhance the production of hCTLA4Ig in transgenic rice cell suspension cultures. When 1 mM sodium butyrate was added in sugar-free media, relative viability was not reduced, while the productivity was improved 1.3-fold. In addition, by supplementing 87 mM sodium pyruvate as an alternative energy source during the production phase, death rate of the cells was decreased. When sodium pyruvate was not added, most cells became dead at day 6. However, by adding sodium pyruvate, 18% of viability can be maintained until day 10 and the production of hCTLA4Ig was enhanced 1.4-fold. When the combination of sodium pyruvate and sodium butyrate at optimum concentrations was added, the highest viability and hCTLA4Ig production could be obtained. The highest level of hCTLA4Ig reached up to 35 mg/L at day 10.

• 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.

• 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.

• IMMUNE NETWORK
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• v.3 no.4
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• pp.302-309
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• 2003

Background: CTLA4 (CD152), which is expressed on the surface of T cells following activation, has a much higher affinity for B7 molecules comparing to CD28, and is a negative regulator of T cell activation. In contrast to stimulating and agonistic capabilities of monoclonal antibodies specific to CTLA-4, CTLA4Ig fusion protein appears to act as CD28 antagonist and inhibits in vitro and in vivo T cell priming in variety of immunological conditions. We've set out to confirm whether inhibition of the CD28-B7 costimulatory response using a soluble form of human CTLA4Ig fusion protein would lead to persistent inhibition of alloreactive T cell activation. Methods: We have used CHO-$dhfr^-$ cell-line to produce CTLA4Ig fusion protein. After serum free culture of transfected cell line we purified this recombinant molecule by using protein A column. To confirm characterization of fusion protein, we carried out a series of Western blot, SDS-PAGE and silver staining analyses. We have also investigated the efficacy of CTLA4Ig in vitro such as mixed lymphocyte reaction (MLR) & cytotoxic T lymphocyte (CTL) response and in vivo such as experimental autoimmune encephalomyelitis (EAE), graft versus host disease (GVHD) and skin-graft whether this fusion protein could inhibit alloreactive T cell activation and lead to immunosuppression of activated T cell. Results: In vitro assay, CTLA4Ig fusion protein inhibited immune response in T cell-specific manner: 1) Human CTLA4Ig inhibited allogeneic stimulation in murine MLR; 2) CTLA4Ig prevented the specific killing activity of CTL. In vivo assay, human CTLA4Ig revealed the capacities to induce alloantigen-specific hyporesponsiveness in mouse model: 1) GVHD was efficiently blocked by dose-dependent manner; 2) Clinical score of EAE was significantly decreased compared to nomal control; 3) The time of skin-graft rejection was not different between CTLA4Ig treated and control group. Conclusion: Human CTLA4Ig suppress the T cell-mediated immune response and efficiently inhibit the EAE, GVHD in mouse model. The mechanism of T cell suppression by human CTLA4Ig fusion protein may be originated from the suppression of activity of cytotoxic T cell. Human CTLA4Ig could not suppress the rejection in mouse skin-graft, this finding suggests that other mechanism except the suppression of cytotoxic T cell may exist on the suppression of graft rejection.

• Journal of Life Science
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• v.18 no.11
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• pp.1471-1478
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• 2008

In this study, nitric oxide (NO) production in a macrophage-lymphocyte co-culture system was used to assess the cytokine producing capability of cells during endotoxin tolerance in mice. Incubation of peritoneal macrophages with interferon-$\tau$ (IFN-$\tau$) in the presence of lipopolysaccharide (LPS) augmented NO synthesis. Exogenous tumor necrosis factor-$\alpha$(TNF-$\alpha$) could also replace LPS for the stimulation of NO production. Macrophages co-cultured with splenic lymphocytes showed augmented NO synthesis by LPS alone. However, pretreatment of mice with 2.5 mg/kg LPS completely prevented the lethality and the increase of blood TNF-$\alpha$ and IFN-$\tau$ after the second challenge with a lethal dose of LPS. In addition, when macrophages prepared from LPS-tolerant mice were co-cultured with normal splenocytes, LPS also could not induce the production of NO, even in the presence of exogenous TNF-$\alpha$. Moreover, when normal macrophages were co-cultured with splenocytes obtained from LPS-tolerant mice, stimulation with LPS could not evoke the NO production enhancement. However, this down-regulation was able to reverse by exogenous IFN-$\tau$ or concanavalin A (ConA), a stimulator of IFN-$\tau$ production. Our results indicate that not only macrophages but also lymphocytes contribute to LPS tolerance. As INF-$\tau$ can enhance the expression of TNF-$\alpha$, the decrease of INF-$\tau$synthesis from lymphocytes may orchestrate with the decrease of TNF-$\alpha$ synthesis from LPS-tolerant macrophages for the production of tolerant state and the prevention of excessive inflammation. Therefore, LPS tolerance may be exploited for prophylaxis of severe sepsis in patients at risk.

• Korean Journal of Veterinary Research
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• v.16 no.2
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• pp.151-158
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• 1976

The tuberculin test have been widely utilized to diagnose tuberculosis of the dairy cattle. It was reported that approximately 70% of the dairy cattle with positive tuberculin reaction in Korea had been non-pathogenic lesion. So the studies on the specific method to diagnose tuberculosis of them is really need in Korea. Therefore this study investigated upon the diagnostic method for cattle tuberculosis in the aspect of cellular-immunology. The results obtained in this investigation are summarized as follows: 1. All the tuberculin tests to the swine inoculated with BCG.(B group), Mycobacterium avium (A group) and Mycobacterium smegmatis (S group), respectively, were represented high positive reaction and were no differences in the species of inoculated bacteria. 2. In the migration inhibition test using Iymphocyte in circulating blood of the swine inoculated with three species of acid-fast bacteria respectively, the test to A group was represented slight positive for migration in the presence of $15{\mu}g/ml$-PPD and the other reaction were clear and total positive for migration inhibition in the presence of $25{\mu}g/ml$-PPD or more, and the test to B group was the same results as to A group approximately. The test to S group was represented slight positive for migration inhibition in the presence of $15{\mu}g/ml$-PPD, but the results were the same in the presence of $25{\mu}g/ml$-PPD and $35{\mu}g/ml$-PPD. These results showed that there were remarkable differences between group A, B and group S in the presence of $25{\mu}g/ml$-PPD and $35{\mu}g/ml$-PPD. 3. The transformation rates of lymphocyte in PPD treated tissue culture had no significance without any relation between PPD treatment and non-treatment but A group and B group showed significance. And A group and B group showed high significance in comparison with N group and S group in the LSD examination. 4. The infection rated in lymphocyte of BCG inoculated after 3 days tissue culturing were represented those of high infection but its cellular degeneration rates almost did not change. The infection rates in bacilli in N group and S group were low but after 3 days inoculation it shewed higher cellular degeneration.

• KSBB Journal
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• v.24 no.3
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• pp.246-252
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• 2009

Rice cells were transformed to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter. hCTLA4Ig was produced and secreted into culture media inducibly when sugar was depleted. The obstacles of this system are the cell death and release of proteases by sugar starvation. These problems resulted in the losses of stability and productivity of hCTLA4Ig. Therefore, the effects of proline as an inhibitor of cell death were investigated. When 4 mM proline was added in sugar-free media, the cell death and release of proteases were reduced. As a consequence, the production of hCTLA4Ig was enhanced. In addition, the effects of protein stabilizers such as gelling agents were studied. It was found that the application of 0.01 g/L gelatin led to an increase in hCTLA4Ig production. This increase might be originated from the stabilization of hCTLA4Ig. In conclusion, the production of hCTLA4Ig could be enhanced by the additions of proline and gelatin in transgenic rice cell cultures.