• The Journal of the Korean Society for Microbiology
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• v.22 no.2
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• pp.163-166
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• 1987

A new method for rosette assay is described for the detection of antigen-specific lymphocytes from BCG-infected mice using sheep erythrocytes coated with BCG antigen. The optimal concentration of BCG antigen for preparation of indicator cells and the incubation time of antigen coated erythrocytes-lymphocytes mixture were $50\;{\mu}g/ml$ and 1 h, respectively. The number of rosette-forming cells (RFC) during the course of BCG infection showed gradual increase as infection progressed and RFC was reached maximum (about 5-7% of splenic lymphocytes formed rosette) at 3 or 4 weeks after infection.

• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.525-530
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• 1989

In order to enumerate the T-lymphocytes in bovine peripheral blood lymphocytes (PBL) by E rosette assay, KGRBC were treated with various concentrations of 2-aminoethylisothiouronium bromide(AET) and dextran(Dex), singly or in combination. To further standardize the assay, optimum concentration of AET- and/or Dex-treatment and incubation time for rosette forming cell(RFC) counts were determined. The levels of B-lymphocytes in the PBL were evaluated by erythzocyte-antibody($EA_{Fc}$)- and erythrocyte-antibody-complement (EAC)-rosetting techniques. The results obtained were as follows; The PBL from 20 clinically normal Korean cattle were formed as low percentage of spontaneous E-rosette ($6.7{\pm}2.4%$) in control group, whereas in KGRBC treated with 0.1M AET for 20 minutes and 8% Dex were formed as $37.3{\pm}2.7%$ and $45.1{\pm}2.1%$, respectively. And the synergistic effects were noted no less than $66.5{\pm}5.6%$ when the KGRBC treated with 0.1M AET and 8% Dex subsequently and rate of RFR did not change significantly between 3~24 hours incubation time at $4^{\circ}C$, EA-and EAC-RFR were $23.3{\pm}9.1%$ and $23.1{\pm}7.9%$, respectively. These results suggest that the KGRBC would be a useful agent for the enumeration of T-lymphocytes by E rosette assay and B-lymphocytes by EA- or EAC-rosette assay in cattle-PBL.

• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.493-496
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• 2016

Peach rosette mosaic virus (PRMV) is a plant virus that was first reported in 1933 by Peach. It can infect hosts including peach, grape, blueberry, dandelion, plum, cherry tree, and weeds. PRMV is non-reportable in Korea, but it is designated as a controlled virus requiring plant quarantine. In this study, for the rapid and specific detection of PRMV, we developed an assay using loop-mediated isothermal amplification (LAMP). Comparison between conventional polymerase chain reaction (PCR) methods (real time-PCR and nested PCR) and LAMP for the detection of PRMV revealed an equivalent level of sensitivity by all the tested methods. For the LAMP assay, outer primer sets were used to amplify a 264-bp PCR product, which was then digested using the restriction enzyme Pvu II (CAG/CTG), and the visualization of two digestion fragments (207 + 57 bp) indicated a positive reaction. The developed LAMP assay for PRMV is expected to enable the rapid monitoring of PRMV in plants.

• Korean Journal of Veterinary Research
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• v.32 no.2
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• pp.175-179
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• 1992

To develope the methods for isolation and enumeration of lymphocyte subpopulations in pigs, we carried out the rosette-assay using sheep erythrocytes(SRBC) and Korean native goat erythrocytes(GRBC) as a target cells. To enumerate T lymphocytes, E-rosette methods were applied with RBC treated with various concentration of polymers such as Aet and Dex, singly or in combination. And to enumerate B lymphocytes, EAand EAC-rosette assay was adopted. The results were as follows; 1. E-RFR with polymer-untreated SRBC and GRBC was $32.9{\pm}7.9%$ and $31.3{\pm}9.4%$, respectively. On the other hand, RFR with 0.1M Aet plus 8% Dex treated SRBC and GRBC was increased about two-fold($67.8{\pm}7.4%$ and $69.8{\pm}8.5%$), respectively. 2. EA-RFR with SRBC and GRBC were $ 39.1{\pm}10.2%$ and $32.6{\pm}6.1%$, respectively. 3. EAC-RFR with SRBC and GRBC were $27.6{\pm}7.0%$ arld $21.0{\pm}3.2%$, respectively. These results showed that both SRBC and GRBC could be recommanded as a binding cells for rosetteassay to isolate of lymphocyte-subpopulations in pigs.

• The Journal of Internal Korean Medicine
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• v.28 no.2
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• pp.321-332
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• 2007

Objectives : Anticancer and immune-modulating effects of several Korean medical prescriptions including Yukgunja-tang, Bohwa-tang, Sogam-Won, and Kamiboa-tang were investigated. Methods : In vitro anti-cancer effects were measured by cytotoxicity MTT assay using SNU-1 gastric cancer cell lines, In vivo anti-cancer effects were measured by increased life span of S-180 sarcoma-injected ICR mouse. Immune-modulating effects were analyzed by measuring hemagglutinin titer, appearance of rosette forming cells, lymphocyte proliferation, and phagocytic index in methotrexate-pretreated mice. Results : In vitro assay showed that only Sogam-won showed cytotoxic effect with $IC_{50}$ of 87.9 ${\mu}g/ml$. All other prescriptions showed no cytotoxic effects against SNU-1 gastric cancer cell line. However, in vivo assay showed that Sogam-won showed lowest anti-cancer effects in contrast to its highest cytotoxic effects, Kamiboa-tang, which showed no cytotoxic effect, showed the highest in vivo anticancer effects, with increased life span of 140%. Kamiboa-tang showed significant immune-enhancing activities by significantly increasing rosette forming cells, lymphocyte proliferation, and phagocytic index in methotrexate-pretreated mice (P<0.05). Conclusion : The anticancer effect of Kamiboa-tang is not mediated by direct inhibition of cancer cells but is mediated by improving immune reactions against cancer cells.

• Korean Journal of Veterinary Research
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• v.27 no.2
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• pp.253-258
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• 1987

The erythrocytes(E) rosette forming capacity of peripheral blood lymphocytes(PBL) in Korean native cattle was determined with 2-aminoethylisothiouronium bromide hydrobromide(AET) and dextran(Dex)-treated sheep erythrocytes(SRBC). To further standardize the assay, optimum concentration of AET and/or Dex-treatment and incubation time for rosette forming cell(RFC) counts were determined. In untreated SRBC resuspended in the rosetting medium(RPMI 1640 containing 10% FCS), PBL from 7 animals formed low percentage of rosettes($7.8{\pm}6.0%$). Both AET and Dex treatment not only enhanced the rosette formation but also made it easy to enumerate rosettes by increasing numbers of SRBC attached on them. SRBC treated with 0.1M AET for 20 min or 8% Dex formed the highest percentages of rosettes, ($35.7{\pm}6.0%$ and $48.3{\pm}4.7%$, respectively) and were used in subsequent studies. With SRBC treated with 0.1M AET for 20 min and suspended in 8% Dex, the maximum RFC observed were $56.6{\pm}6.8%$ in female and $65.8{\pm}6.3%$ in male and the rates of RFC did not change significantly between 3 and 20 hr incubation time at ${4^{\circ}C}$.

• The Korean Journal of Food And Nutrition
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• v.10 no.4
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• pp.533-538
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• 1997

This study was conduced to investigate on immune response of the hot water extract(PHE), 50% methanol extract(PME) and acetone extract(PAE) from Glycyrrhiza glabra. The experiment was carried out by phagocytosis, plaque forming cell(PFC), hemalysin titration(HY) and rosette forming cell(RFC) assay by using BALB/c mice. The results obtained from this study are as follow ; The effects of Glycyrrhiza glabra extracts on phagocytosis was tended to be slight increase in GME and GAE groups compared to the control group, but not significant. In the experiment of PFC and HY, the results of experiment groups which was given each samples were significantly higher than the control group. The result of rosette forming cell in GME and GAE groups were significantly higher than control group.

• YAKHAK HOEJI
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• v.40 no.5
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• pp.599-607
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• 1996

Acetylarsonate was prepared for testing antitumor and immunological effects. It showed cytotoxicity directly on Sarcoma 180. L1210 and MOLT-4 by MTT assay. It did not seemed to trigger the mitosis of human lymphocytes in culture, but that showed the cytotoxicity with higher dose. The rosette formation and spleen weight of mouse which acetylarsonate was administered to for 2 weeks were increased. Furthermore, peripheral helper T- and cytotoxic/suppressor T-lymphocytes were increased in acetylarsonate-injected-mice significantly when it was estimated with simultaneous 2 color analysis using anti Lyt2-FITC and L3T4-PE monoclonal antibody by Flow cytometer.

• Archives of Pharmacal Research
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• v.15 no.1
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• pp.20-29
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• 1992

Effects of squalene on cellular and non-specific immune responses and antitumor activity in mice were investigated. Cellular and non-specific immunological assay parameters adopted in the present study were delayed-type hypersensitivity reaction and resette forming cells (RFC) for cellular immunity, activities of natural killer (NK) cells and phagocyte for non-specific immunity. Squalene resulted in marked increases of cellular and non-specific immune functions and enhancement of host resistance to tumor challenge in dose-dependent manner.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.6 no.1
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• pp.47-65
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• 2000

In order to investigate antitumor and immune response effect by Hyangsapyungwisan after Sarcoma-180 cells and methotrexate were treatred each other, the extract of Hyangsapyungwisan was orally administered to ICR mice for 14 days. To evaluate the effects of the Hyangsapyungwisan, 50% inhibition concentration($IC_{50}$), mean survival days, tumor weight for antitumor effects, hemagglutinin titer, hemolysin titer, rosette forming cells, natural killer cell activity and productivity of interleukin-2 for immune responses measured in ICR mice. The results were summarized as follows: 1. Mean survival time in Hyangsapyungwisan-treated group was slightly prolonged, as compared with control group(13.46%). 2. On the MTT assay, cell viability was significantly inhibited by $5{\mu}g/well,\;2.5{\mu}g/well,\;1.25{\mu}g/well,\;and\;0.625{\mu}g/well$ of Hyangsapyung-wisan concentration inhibited cell viability significantly. $IC_{50}$ for cell viability was $11.59{\mu}g/well$. 3. Tumor weight in Hyangsapyungwisan treated group was depressed, as compared with the control group(p<0.05). 4. Hemagglutinin titer in Hyangsapyungwisan-treated group was slightly increased with no significance, as compared with the control group. 5. Hemolysin titer in Hyangsapyungwisan-treated group was silightly increased, as compared with the control group(p<0.05). 6. Rosette forming cells in Hyangsapyungwisan-treated group was silightly increased, as compared with the control group(p<0.05). 7. Naural killer cell activity in Hyangsapyungwisan-treated group was significantly increased(p<0.05). 8. Production of interleukin-2 was significantly increased(p<0.05). According to the above results, Hyangsapygwisan had prominent antitumor effects, and enhance both cellular and humoral immunity in mice.