• 대한한방소아과학회지
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• 제13권1호
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• pp.253-275
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• 1999

The effects of Sagunjatang and Samultang on the immunosuppression induced by methotrexate(MTX) in rats were investigated in this study. The multiple parameters of immunity assessed in each rats included leukocyte count, lymphocyte rate, the number of lymphocyte in tibial bone marrow, contact hypersensitivity to DNFB, morphological change of thymocyte and IgG antibody on SDS-PAGE. Sprague-Dawley male rats were used and divided into five groups at random. Group A was normal control. Group B, the MTX treatment control, was injected i.v. with 2mg/kg of on days 9, 11 after sensitization with SRBC on 5th day. Group C, the experimental control, was treated Sagunjatang for 18days and MTX. Group D was treated Samultang for 18days and MTX. Group E was treated Sagunjatang-Samultang for 18days and MTX. The dosage of Sagunjatang and Samultang was $1m{\ell}/day$ respectively. In the case of Group E, rats Were fed Sagunjatang $1m{\ell}$ in the morning and Samultang $1m{\ell}$ in the afternoon. The results are summarized as follows: 1. Leukocyte count in rats induced by intravenous sensitization with SRBC was decreased significantly in Group E. 2. Leukocyte counts of 2weeks later after being treated MTX were increased significantly in Groups C and D. 3. Lymphocyte rate in rats induced by intravenous sensitization with SRBC wasn't changed significantly in all the experimental groups. 4. Lymphocyte rate of 2weeks. later after being treated MTX was increased significantly in Group D. 5. The number of lymphocyte in tibial bone marrow was incereased significantly in Group C. 6. Contact hypersensitivity wasn't changed significantly in all the experimental groups. 7. Morphological finding of thymocyte in group C was similar to normal group as compared with control group. 8. Purified IgG of all the experimental groups showed two bands of 50,000 and 25,000 on SDS-PAGE. But there was no difference among experimental groups.

• 약학회지
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• 제35권3호
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• pp.154-164
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• 1991

These experiments were conducted to investigate the effects of Glycyrrhizae Radix extract(GR) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [$^{3}$H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}Ca^{2+}$ to P388D$_{1}$ cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GR(10$^{-3}$g/ml). Histamine production in mouse spleen cell culture was significantly increased by 48 hour incubation added 0.25$\mu\textrm{g}$/ml of Con A. Con A-dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 $\mu\textrm{g}$/ml of Con A. GR depressed histamine contents at 10$^{-9}$~10$^{-4}$g/ml. and Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-5}$~10$^{-4}$g/ml. IL-1 activity was significantly decreased by 10$^{-8}$~10$^{-4}$g/ml of GR. $Ca^{2+}$ uptake was not changed by GR, but antibody production markedly increased at 10.0~50.0 mg/kg of GR. From the above results, it is suggested that GR have immuno-regulatory action; GR decreased cell-mediated immune response and increased antibody production by B lymphocyte at high doses.

• 한국수의병리학회지
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• 제7권1호
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• pp.11-15
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• 2003

Antigen retrieval (AR) techniques were widely used to recover the antigenicity from the fixed tissues, which were guided by the philosophy of rendering immunohistochemistry (IHC) applicable to routine formalin-fixed, paraffin-embedded tissues for wide application of IHC in research and clinical filed for morphological observation like as anatomy, histology and pathology. Protease antigen recovery (PAR) is an AR technique, which is obtained the antigen retrieve by using enzyme digestion, and commonly used in IHC field. However, during the IHC for the detection of ovine herpesvirus 2 (OvHV-2) antigen, we noted lymphocyte-like cells-specific staining in the infiltrated cells into various organs like as liver and kidney, which was also shown in the IHC tissues with isotype control. However, those signals were not observed in the tissues conducted with in situ hybridization. Therefore, we analyzed the specificity of the IHC detection results. We found that PAR may induce false-positive result during IHC in lymphocyte-like cells, which were infiltrated mainly around vessels and in interstitial tissues. Through the Phenotyping, we realized that those false-positive cells were B-cell-related cells. These results suggest that PAR, a AR using protease, may induce non-specific false-positive reactions during IHC.

• Environmental Analysis Health and Toxicology
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• 제13권1_2호
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• pp.33-41
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• 1998

Chlorpyrifos, o,o diethyl-o-(3,5,6-trichloro-2-pyridyl) phosphorothioate, is a broad spectrum organophosphate insecticide. The use of chlorpyrifos has been increased more and more as pesticide. But the effects of chlorpyrifos on the immune alterations has not been yet observed. Therefore, we investigated the effects of chlorpyrifos on the immune alterations in CICA male mice. Chlorpyrifos was administered to mice by a single intraperitoneal injection for the purpose of observing acute effects. On the one hand to get the information on immunopathologic alterations we observed hematological values, counted total circulating leukocytes and assessed the ratio of lymphocytes and neutrophils from the peripheral blood, measured the ratio of organ/body weight and counted splenic cellularity in CBA male mice which treated chlorpyrifos intraperitoneally. But we could not find any significant immunopathologic alterations statistically by a single intraperitoneal injection. Also, the exposure of chlorpyrifos caused no significant change in the number of PFC/10$^6$ spleen cells at any three given doses. On the other hand a singte intraperitoneal injection of chlorpyrifos decreased the lymphocyte proliferation response slightly to ConA or LPS stimulation at a dose of 6 mg/kg b.w. Administrations of chlorpyrifos reduced mixed leukocyte response(MLR). MLR was decreased moderately at doses of 3mg/kg b.w. and 6mg/kg b.w. Therefore, all these findings suggest that chlorpyrifos may alter the immune functions acutely. especially by the changes of T lymphocyte activity.

• 한국어병학회지
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• 제6권2호
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• pp.111-122
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• 1993

B 세포의 $V_H$ 유전자가 어떠한 기작으로 선택되어지는 지는 현재 명확히 밝혀져 있지 않다. 본 연구에서는 transformation 등의 방법에 의한 편향된 분석결과를 피하고자 in situ hybridization 기법을 이용하여 정상적인 single 세포가 발현한 $V_H$ 유전자를 분석하였다. $V_H$ 유전자간에 나타나는 DNA 배열의 유사성 때문에 in situ 기법에서 가장 중요한 것은 probe 농도와 세척 stringency의 결정이다. LPS-stimulated된 spleen B 세포에 대해서 $C{\mu}$와 $V_HJ558$ $^{35}S$-RNA probe는 $2{\sim}4{\times}106cpm$/slide의 농도에서 낮은 background와 적정수의 positive 세포를 관찰할 수 있었으며 세척조건으로서는 $54^{\circ}C$에서 40~50%의 formamide를 사용할때 최적이라는 것을 $C{\mu}$, $V_{H}S107$, 그리고 $V_{H}J558$ probe를 이용한 실험에서 결정하였다. 위의 조건하에서 spleen B 세포가 발현한 $V_H$ 유전자를 분석하여 본 결과 각각의 $V_H$ gene family 발현 빈도는 각각의 family 크기에 비례하여 결정된다는 것을 알 수 있었다. 이러한 결과들은 여러 다른 발달 단계에 있는 bone marrow B 세포에 대해서도 동일한 결과를 보여 주어 어떤 특수 $V_H$ gene family의 발현이 B 세포의 발달단계에 따라 특이하게 변화하는 것은 아니라는 것을 나타내 보여 주었다. 그러므로 $V_H$ 유전자의 이용은 B 세포가 differentiation하는 것과는 무관하게 무작위 적으로 선택되어 진다는 것을 밝혔다.

• IMMUNE NETWORK
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• 제1권2호
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• pp.126-134
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• 2001

Background : Paeonia japonica Miyabe is a medicinal plant which has been widely used as a component of blood-building decoctions (Chinese medicinal concept : Bu-Xie). The immunopharmacological characteristics of the extract of Paeonia japonica (PJ) were investigated. Methods : The effects of fractions of PJ extract on lymphocyte proliferation were measured by $H^3$-thymidine incorporation assay. The proliferated lymphocyte subsets were analyzed in flow cytometry. The subset cell populations of spleen cells were separated by magnetic cell separation system, and their proliferation by the extract were investigated. The effect of the extract on antibody production was determined in mice challenged with sheep red blood cells (SRBC) using hemolytic plaque forming cell assay. Results : Spleen cells were proliferated by water extract of PJ. Polysaccharide fraction (PJ-P) of the extract was most active in the proliferation. It was found in flow cytometry that the lymphocyte subset proliferated by PJ-P was B cell population. Among the separated subset cell populations, T cell-depleted cell population and macrophage-depleted cell population were most proliferated by PJ-P. However, positively selected populations of B cells and T cells were not proliferated by PJ-P. These results indicate that B cell proliferation by PJ-P may require the assistance of macrophages or T cells. These results suggest that firstly PJ-P may stimulate macrophages or T cells, and then B cells are activated. The number of antibody-secreting cells was increased by administration of PJ-P in mice immunized with SRBC as a T-dependent antigen. Conclusion : These results suggest that macrophages and accessory cells are directly activated by PJ-P and then helper T cells and B cells are indirectly activated. As the results, immune responses might be coordinately improved. In conclusion, PJ-P, a polysaccharide of P. japonica, may be a characteristic immunostimulator, which is analogous to polysaccharides such as lentinan, PSK and ginsan.

• Restorative Dentistry and Endodontics
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• 제20권1호
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• pp.235-249
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• 1995

This study was designed to identify the lymphocytes present and to examine the relation between lymphocytes population and clinical symptoms of the pulps clinically diagnosed as normal and irreversible pulpitis. We recorded the history and severity of the pain and performed several clinical tests, before extirpation of vital, irreversibly inflamed pulps in routine endodontic treatment. Then the teeth were divided into two groups. Five teeth, categorized in acute symptom group, had severe spontaneous pain, particularly at night and were extremely sensitive to cold and heat. The other 15 teeth with history of mild to moderate pain and with or without cold or heat responses were categorized as chronic symptom group. Inflamed pulps were also classified into 8 minor groups by presence or absence of signs or symptoms related to the involved teeth, including the presence of pain on percussion, pain on heat and cold stimuli and the periodontal pocket depth. All extirpated pulps were immediately immersed in ultra low-temperature freezer($-74^{\circ}C$), and they were sectioned $6{\mu}m$ in thickness. Specimens were stained using three-stage indirect immunoperoxidase techniques(DAKO, LSAB kit) and monoclonal antibodies for detecting the presence of T lymphocytes(T), B lymphocytes(B) and helper(T4) and suppressor(T8) lymphocytes. Following results were obtained; 1. All the examined normal and inflamed pull) tissues had positive staining for T lymphocytes and T helper and T suppressor cells. But B cells were observed only in inflamed pulp. 2. Statistically more T and B cells were observed in acute symptom group as compared with chronic symptom group(p<0.05). 3. Cell ratio of BIT in acute symptom group were significantly higher than that of chronic symptom group(p<0.05). 4. Only B cells were significantly increased in the percussion positive group than the number of B cells in percussion negative group(p<0.05). 5. No differences were observed in the number of different cell types among other minor groups.

• Asian-Australasian Journal of Animal Sciences
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• 제18권6호
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• pp.879-883
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• 2005

The integral relationship between the occurrence of lymphocyte nuclear pockets (LNPs) and BLV-infection was examined in Holstein-Friesian dairy cattle in Korea. Transmission electron microscope (TEM) was used to detect LNP in peripheral blood lymphocytes. Morphologically, the membranes of LNP were composed of two layers of double nuclear membrane. The full thickness of LNP membranes including inner and outer nuclear membrane was 60 to 70 nm. LNP prevalence was different according to the bovine leukemia virus (BLV) infection status; in BLV-seropositive cattle, LNP prevalence was 48.4% and in BLV-seronegative cattle prevalence was 5.9%. Moreover, even in seropositive animals, leukemic group was the highest at 70% positive among the groups, followed by suspect group (42.4%) and aleukemic group (23.1%). Consequently, the numbers of LNP were increased in proportion to increase of the numbers of leukocytes among BLV-seropositive cattle. The numbers of LNP per lymphocyte were increased in BLVseropositive cattle compared with seronegative cattle. The mean numbers of LNP per 100-lymphocytes were 0.35, 0.77, 1.64 and 4.7 in BLV-seronegative, BLV-seropositive aleukemic, suspect and leukemic groups, respectively. Thus, it is reasonable that LNP test can be used as the one of the diagnostic criteria of BLV infection.

• Journal of Microbiology and Biotechnology
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• 제9권4호
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• pp.398-403
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• 1999

In our previous studies, we reported that hybridoma B-lymphocytes can be used to determine the virulence of Listeria species in an in vitro cytotoxicity assay. Here, we examined the cytopathic effect, i.e., membrane damage and the nature of cell death induced by Listeria monocytogenes on murine hybridoma B-lymphocytes (Ped-2E9). Membrane damage was assessed by microscopic analyses and by measuring the release of intracellular alkaline phosphatase(AP) and lactate dehydrogenase (LDH). Cell death was determined by DNA fragmentation analyses using agarose gel electrophoresis. Infection by listeriolysin O (LLO)-producing L. monocytogenes strains induced substantial amounts of AP and LDH release from Ped-2E9 hybridoma B-cells, suggesting severe membrane damage in these cells, while an LLO-negative L. monocytogenes mutant strain had no effect. An LLO-producing recombinant L. innocua ($prifA^+hly^+$) strain also induced high AP and LDH release and cytopathic changes in Ped-2E9 cells. Light or scanning electron microscopic examination revealed L. monocytogenes mediated membrane destabilization, pore formation, intense cytoplasmic granulation, bleb formation, and lysis of Ped-2E9 cells. LLO-producing L. monocytogenes and L. innocua ($prifA^{+}hly{^}+$) also induced ladder-like DNA fragmentation in Ped-2E9 cells. Collectively, these results suggest that L. monocytogenes, specifically LLO-producing strains, can induce a severe cytopathic effect leading to apoptosis in hybridoma B-lymphocytes (Ped-2E9).

• Childhood Kidney Diseases
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• 제19권2호
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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.