• The Korean Journal of Nuclear Medicine
• /
• v.39 no.1
• /
• pp.26-33
• /
• 2005

Purpose: The purpose of this study is to evaluate migration of technetium-99m hexamethylpropylene amine oxime ($^{99m}Tc$-HMPAO) labeled immature and mature dendritic cells (DC) in the mouse. Methods: DC were collected from bone marrow (BM) of tibiae and femurs of mice. Immature and mature DC from BM cells were radiolabeled with $^{99m}Tc$-HMPAO. To evaluate the functional and phenotypic changes of DC from radiolabeling, the allogeneic mixed lymphocyte reaction (MLR) and fluorescence-activated cell sorting (FACS) analysis were performed before and after labeling with $^{99m}Tc$-HMPAO. Migration of intravenously injected DC (iv-DC) was assessed by serial gamma camera images of mice with or without subcutaneous tumor. Percent injected dose per gram (%ID/g) was calculated in lungs, liver, spleen, kidneys, and tumor through dissection of each mice after 24 hours of injection. Results: Labeling efficiency of immature and mature DC were $60.4{\pm}5.4%\;and\;61.8{\pm}6.7%$, respectively. Iv-DC initially appeared in the lungs, then redistributed mainly to liver and spleen. Migration of mature DC to spleen was significantly higher than that of immature DC ($38.3{\pm}4.0%\;vs.\;32.2{\pm}4.1%$ in control group, $40.4{\pm}4.1%\;vs.\;35.9{\pm}3.8%$ in tumor group; p<0.05). Migration to tumor was also significantly higher in mature DC than in immature DC ($2.4{\pm}0.3%\;vs\;1.7{\pm}0.2%$; p=0.034). Conclusion: Assessment of migration pattern of DC in mice was possible using $^{99m}Tc$-HMPAO labeled immature and mature DC. Migration of mature DC to spleen and tumor was higher than that of immature DC when they were i.v. injected.

• The Korean Journal of Food And Nutrition
• /
• v.11 no.1
• /
• pp.107-113
• /
• 1998

This experiment has conducted to evaluate whether single injection of red ginseng extract including 50% ethanol extract, crude saponin, and lipid soluble fraction can induce oxidative burst of mouse peritoneal macrophages with use of fluorescence spectrophotometer. To optimize conditions of fluorescent spectrophotometry, concentrations of DCFH-DA(2', 7' -dichlorofluorescin diacetate) was 1.6 ${\mu}{\textrm}{m}$ and control oxidative burst by Zymosan A and PMA(phorbol myristate acetate) were 100$\mu\textrm{g}$, 250ng, respectively. Though in vitro macrophages failed to induce increment of H2O2 production, but 50% ethanol extract group induced significant enhancement of H2O2 production when zymosan A triggered oxidative burst. On the other hand, lipid soluble fraction enhanced significantly H2O2 production than that of control group. These findings consisted with the other reports which showed ginsenosides inhibited nitric oxide production and lipid soluble fraction activated colony stimulating factor(granulocyte - monocyte) activity in bone marrow stem cells. As is well known, lipid soluble fraction contains phenol compound, polyacetylene compound and alkaloids. Further study would unravel which component of it can induce H2O2 production of macrophages. Key words : Red ginseng(Panax ginseng), H2O2 production, macrophages.

• IMMUNE NETWORK
• /
• v.10 no.6
• /
• pp.239-246
• /
• 2010

Background: Monoclonal antibodies (mAbs) recognizing Class III epitope of CD34 are essential for flow cytometric diagnosis of leukemia. Methods: 27H2 mAb was developed from a mouse alternatively immunized with human acute leukemia cell lines, KG1 and Molm-1. Using flow cytometric analysis of various leukemic cell lines and peripheral blood, immunohistochemical study of frozen tonsil, we characterized 27H2 mAb. Antigen immunoprecipitated with 27H2 mAb immunobloted with anti-CD34 mAb. A case of bone marrow sample of acute lymphoblastic leukemia (ALL) patient was obtained at CBNU Hospital. For epitope identification enzyme treatment with neuraminidase and O-sialoglycoprotein endopeptidase (OSGE) and blocking assay with known classIII mAb (HPCA-2) were done. Results: Only KG1 and Molm-1 revealed positive immunoreactivity. Immunohistochemical staining disclosed strong membranous immunoreactivity on high endothelial venules. Antigen immunoprecipitated by 27H2 mAb showed approximately 100 kDa sized band immunoblotted with anti-CD34 under non-reducing conditions. Epitope recognized by 27H2 mAb disclosed resistancy to both neuraminidase and OSGE treatment and completely blocked with known class III mAb preincubation. CD34 positive leukemic cells in BM of pre B cell ALL patient detected by FITC-conjugated 27H2 and HPCA-2 were identified with similar sensitivity. Conclusion: A novel murine mAb recognizing class III epitope of human CD34 with high affinity, which is useful for flow cytometric diagnosis of leukemia, was developed.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.35 no.1
• /
• pp.28-34
• /
• 2006

The ethanol extracts of Puerariae Radix inhibited cyclooxygenase-2 (COX-2) activity in bone marrow derived mast cells (BMMC). COX-2 is responsible for the production of large amounts of proinflammatory prostaglandins (PGs) at the inflammatory site. We have investigated the anti-inflammatory effect of ethyl acetate fraction from $70\%$ ethanol extract of Puerariae Radix (EPR), and attempted acetic acid induced writhing to verify the analgesic effect. Inflammation was induced by interleukin-1 (IL-1), tumor necrosis factor-a (TNF-a), $inteferon-\gamma$ $(IFN-\gamma)$ and lipopolysaccharide (LPS). EPR showed strong inhibitory efficacy against cytokine-induced proteoglycan degradation, prostaglandin $E_2\;(PGE_2)$ production, nitric oxide (NO) production, and matrix-metalloproteinases (MMPs) expression in mouse macrophage and rabbit articular chondrocyte. In the writhing test, EPR $(200\~400\;mg/kg)$ exhibited a dose-dependent inhibition of writhing. The results indicate that EPR have anti-inflammatory and analgesic activities, and could be a good herbal medicine candidate for treating of osteoarthritis (OA).

• Clinical and Experimental Pediatrics
• /
• v.52 no.7
• /
• pp.824-831
• /
• 2009

Purpose : We aimed to investigate the efficacy of and functional recovery after intracerebral transplantation of different doses of mouse mesenchymal stem cells (mMSCs) in immature rat brain with hypoxic-ischemic encephalopathy (HIE). Methods : Postnatal 7-days-old Sprague-Dawley rats, which had undergone unilateral HI operation, were given stereotaxic intracerebral injections of either vehicle or mMSCs and then tested for locomotory activity in the 2nd, 4th, 6th, and 8th week of the stem cell injection. In the 8th week, Morris water maze test was performed to evaluate the learning and memory dysfunction for a week. Results : In the open field test, no differences were observed in the total distance/the total duration (F=0.412, P=0.745) among the 4 study groups. In the invisible-platform Morris water maze test, significant differences were observed in escape latency (F=380.319, P<0.01) among the 4 groups. The escape latency in the control group significantly differed from that in the high-dose mMSC and/or sham group on training days 2-5 (Scheffe's test, P<0.05) and became prominent with time progression (F=6.034, P<0.01). In spatial probe trial and visible-platform Morris water maze test, no significant improvement was observed in the rats that had undergone transplantation. Conclusion : Although the rats that received a high dose of mMSCs showed significant recovery in the learning-related behavioral test only, our data support that mMSCs may be used as a valuable source to improve outcome in HIE. Further study is necessary to identify the optimal dose that shows maximal efficacy for HIE treatment.

• Journal of Preventive Medicine and Public Health
• /
• v.27 no.1 s.45
• /
• pp.11-24
• /
• 1994

The studies reported here were undertaken to investigate the effects of mercury chloride on immune system of Balb/c mouse employing a flexible tier of in vitro and in vivo assays. Mercury chloride inhibited the proliferative responses of spleen cells to lipopolysaccharide, pokeweed mitogen, and phytohemagglutinin as a dose-dependent manner. This inhibitory effect was observed not only when $HgCl_2$ was added 2nd or 3rd day of 3 days culture period but also when spleen cells was pretreated with $HgCl_2$ for 2 hours. Mercury chloride, however, potentiated the production of IgM and IgG from spleen cells. During the $HgCl_2$ administration by drinking for 3 weeks, the weight gain of mice was significantly blunted than that o control group mice, while no overt signs related to mercury toxicity were noted in any mice of experimental group. There was no change in thymus and spleen weights, and in histological findings of kidney, bone marrow of femur, thymus, spleen, and popliteal lymph node after 3 weeks of mercury exposure. However, $HgCl_2$ induced a significant increase of total serum IgM, IgG including $IgG_1,\;IgG_{2a}\;and\;IgG_{2b}$, and IgE in Balb/c mice. Treatment in vivo with anti-IL-4 monoclonal antibody significantly abrogated the $HgCl_2$-induced increase in total serum IgG1 and IgE. Whereas $HgCl_2$ potentiated total serum IgM and IgG, there was no difference in total serum hemagglutinin to SRBC (Sheep Red Blood Cell) between experimental and control group mice when these mice were immunized with SRBC. All these findings observed in Balb/c mice suggest that mercury perturbates well-orchestrated regulation of immune responses before developing histopathological changes in lymphoid tissues.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.34 no.6
• /
• pp.805-813
• /
• 2005

In our previous study, a novel herb mixture (HIM-I) of Angelim gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix was developed to protect the intestinal and immune systems and promote its recovery against radiation damage. In this study, a new herbal preparation (HemoHIM) with the high immune modulating activity was developed from HIM-I. HIM-I was fractionated into ethanol fraction (HIM-I-E) and polysaccharide fraction (HIM-I-P). And HemoHIM was prepared by adding HIM-I-P to HIM-I. The protective activities against $\gamma$ -irradiation were compared among HemoHIM, HIM-I and the fractions. HemoHIM and HIM-I significantly decreased the radiation-induced DNA damage in vitro, and scavenged hydroxyl radicals in a dose-dependent manner. HemoHIM showed similar activity to HIM-I. In vitro proliferation assay with mouse lymphocytes and bone marrow cells showed that HIM-I-P was remarkably higher than HIM-I and HIM-I-E in cell proliferating activity. HemoHIM showed higher activity than HIM-I and this might be associated with the higher polysaccharide content. The in vivo protective effects of HemoHIM and HIM-I were investigated in $\gamma$-irradiated mice. HemoHIM increased the surviving intestinal crypts to a similar extent compared with HIM-I. In contrast, HemoHIM appeared to be more effective than HIM-I in endogenous spleen colony formation assay. The recovery of white blood cells and lymphocytes in irradiated mice were significantly enhanced by the administration of HemoHIM. Also HemoHIM administration prolonged the survival of irradiated mice. These results showed that the novel herbal preparation, HemoHIM, effectively protected the self-renewal tissues and immune system, and promoted the survival of irradiated mice. Moreover, in comparison with HIM-I, HemoHIM maintained similar activity in the reduction of oxidative damage of self-renewal tissue but exhibited the higher activity in protection and proliferation of immune and hematopoietic cells. These results suggested that HemoHIM might be more effective than HIM-I in immune modulation as well as radioprotection.