• Title/Summary/Keyword: Mononuclear

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Protective Effect of Panax ginseng on TCDD-induced Liver Toxicity in Male Guinea Pigs

  • Hwang, Seock-Yeon;Wee, Jae-Joon;Kim, Si-Kwan;Youn, Nae-Young
    • Biomedical Science Letters
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    • v.9 no.1
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    • pp.51-58
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    • 2003
  • This study was carried out to investigate the preventive and therapeutic effect of Panax ginseng water extract (PG-WE) on the toxicity induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), one of the most notorious toxic environmental pollutants belonging to the group of polyhalogenated aromatic hydrocarbons. Normal control (NC) group guinea pigs (180~200 g) received vehicle and saline, and TCDD-treated (TT) group was given TCDD and saline. P100 and P200 group animals received PG-WE for 28 days since 1 week before TCDD exposure at daily doses of 100 mg/kg b.w. and/or 200 mg/kg b.w., respectively. C100 and C200 group received PG-WE for 14 days starting 1 week after TCDD-exposure. Toxicity was induced by a single intraperitoneal injection of TCDD (1 $\mu\textrm{g}$/kg b.w.). Abnormal increase in AST and ALT activities in TT group was significantly improved by the administration of PC-WE. Microscopically, there were mild to moderate swelling of hepatocytes, hyperchromatism of individual cells, acidophilic cytoplasm and cytoplasm vacuolation of some hepatocytes, slight to moderate variations of staining density, occasional single cell necrosis, variable size and shape of some hepatocytes, small groups of degenerating hepatocytes surrounded by mononuclear cells, dilated sinusoids of centrilobular zone and some loss of lobular architecture in TT group liver. From these results, we could find the protective and therapeutic role of PG-WE in TCDD-induced liver toxicity by examining the blood chemical parameters and histopathological observation.

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Mycobacterium tuberculosis Derived Epitope Peptide Specific CD8+T Cell Responses in Tuberculous Pleurisy

  • Cho, Jang-Eun;Kim, Young-Sam;Park, Moo-Suk;Lee, Kyung-Wha;Lee, Eun-Hee;Cho, Sang-Nae;Cho, Sung-Ae
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.325-332
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    • 2007
  • Cell-mediated immune response (CMI) is a major immune protective mechanism against tuberculosis (TB) infection. Among several components involved in CMI, recent studies suggest that CD8+ T cells are important in controlling TB infection. In our previous report, we defined four Mycobacterium tuberculosis (MTB) derived epiotpe peptides specific for HLA-A*0201-restricted CD8+ T cells. These four peptides are $PstAl_{75-83}$, $ThyA_{30-38}$, $RpoB_{127-135}$ and $85B_{15-23}$. In this study, these epitope peptides specific CD8+ T cell responses in tuberculous pleurisy were investigated using ex vivo $IFN-\gamma$ elispot assay and intracellular $IFN-\gamma$ staining method. As a result, we observed these epitope peptide specific CD8+ T cell responses are induced in all three patients with tuberculous pleurisy suggesting that CD8+ T cells are involved in protective immune mechanism against MTB infection in tuberculous pleurisy. However, the CMI to mitogens and MTB antigens from pleural fluids of patients with tuberculous pleurisy does not seem to correlate with that from peripheral blood, although the sample size is too small to make any conclusion. In sum, the MHC I restricted CD8+ T cell responses seem to be induced efficiently in the pleural fluids, at the site of TB infection, in which the CMI is actively induced. In addition, these experiments suggest that MHC I restricted CD8+ T cell mediated immune responses are also involved in protective mechanism against MTB infection in extra-pulmonary TB.

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Complete Sequences of HIV-1 in a Korean Long-term Nonprogressor with HIV-1 Infection (장기간 진행하지 않는 인면역결핍바이러스(Human Immunodeficiency Virus, HIV)-1 감염자로부터 분리한 HIV-1의 전체 염기서열 결정)

  • Cho, Young-Keol;Lee, Hee-Jung;Desrosiers, Ronald C.
    • The Journal of Korean Society of Virology
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    • v.29 no.2
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    • pp.107-118
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    • 1999
  • To characterize the molecular nature of human immunodeficiency virus (HIV)-1, we determined the full-length HIV-1 sequences from cultured peripheral blood mononuclear cells (PBMC) of a Korean long-term nonprogressor (LTNP). Without antiretroviral therapy, the individual has maintained CD4+ T counts over $500/{\mu}l$ from 1989 to 1999. Plasma viral RNA copy was 992 U/ml in 1998. Culture supernatant showed positive from culture days 9. A series of 9 overlapping PCR products were amplified from cultured PBMC and cloned About 9.2 kb from R of 5' LTR to R of 3' LTR was determined by automated sequencing. The G-to-A hypermutations were shown throughout the entire region. As a result of G to A hypermutations, premature stop codon was found in integrase coding region. Though there was no recombination between subtypes over all genomes, TATA box in both LTRs was TAAAA which is detected in subtype E instead of TATAA in subtype B. And, there were nucleotide GC insertion between $NF-{\kappa}B$ I and Sp1 III, and duplication of $TCF-1{\alpha}$ in LTR. We could not find any deletion of amino acid in Nef, Gag, Pol and Env gene. This study is the first report on molecular nature of full genomes of HIV-1 isolated in Korea.

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Detection of Resistance Mutation to Lamivudine in HIV-1 Infected Patients (Lamivudine 복용 HIV-1 감염자에게서 내성 돌연변이 검색)

  • Cho, Young-Keol;Sung, Heung-Sup;Lee, Hee-Jung;Kim, Yoo-Kyum;Chi, Hyun-Sook;Cho, Goon-Jae;Kang, Moon-Won
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.2
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    • pp.181-190
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    • 2000
  • To investigate resistance to lamivudine (3TC), we examined the incidence of M184V in 20 HIV-1 patients treated with 3TC for $13.1{\pm}9$ months. Fourteen of 20 patients had been exposed to zidovudine (ZDV) or didanosine (ddI) prior to 3TC therapy. Nested PCR targeting to reverse transcriptase (RT) and direct sequencing were performed for peripheral blood mononuclear cells sampled serially. There were resistance mutations to ZDV in at least 9 patients at baseline, although there was no resistance mutation to 3TC. We could detect M184V in 6 (30%) out of 20 patients. The incidence of M184V increased as the duration of therapy prolongs (13% in samples <12 months; 47% in samples ${\ge}12$ months). The frequency of mutation M184V was higher in patients with previous mutation to ZDV than in patients with wild type. Resistance mutation was not detected in 7 patients. This study shows that resistance to 3TC tends to develop rapidly in patients with baseline mutations or two drugs combination therapy than in those treated simultaneously with triple drugs. This report is the first on resistance to 3TC in Korean AIDS patients.

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Increased Expression of Adhesion Molecules on Human Umbilical Vein Endothelial Cells by Orientia tsutsugamushi Infection (Orientia tsutsugamushi 감염에 의한 사람 제대정맥 내피세포에서 부착분자 발현의 변화)

  • Lee, Eun-Bong;Han, Seung-Hoon;Kim, Sang-Wook;Ihn, Kyung-Soo;Seong, Seung-Yong;Kim, Ik-Sang;Choi, Myung-Sik
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.2
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    • pp.159-169
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    • 2000
  • Scrub typhus is caused by Orientia tsutsugamushi characterized by fever, headache, lymphadenopathy and eschar formation. Infiltration of inflammatory cells around blood vessels and within the affected organs isS known to be pathologic hallmark of the scrub typhus. Recently, expression of adhesion molecules on vascular endothelial cells was implicated as an important pathogenic mechanism in rickettsial disease. This study was performed to examine the expression of adhesion molecules and to investigate its role in the pathogenesis of O. tsutsugamushi infection. The expression of adhesion molecules on human umbilical vein endothelial cells (HUVEC) was measured by flow cytometry and indirect immunofluorescence. Expression of E-selectin, ICAM-1 and VCAM-1 was significantly increased 4 hours after the infection and persisted at least for 24 hours. Expression of those molecules was not induced by killed O. tsutsugamushi. Adhesion of polymorphonuclear cells and mononuclear cells to HUVEC was increased after the infection with O. tsutsugamushi. In conclusion, adhesion molecules are expressed on HUVEC during the infection of live O. tsutsugamushi and those molecules can contribute to the infiltration of inflammatory cells during the infection.

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Comparison of Immunomodualtory Effects of Water-extracted Adenophorae Radix, Liriopis Tuber, Dendrobii Herba, Polygonati Odorati Rhizoma and Polygonati Rhizoma (보음약인 사삼, 맥문동, 석곡, 옥죽, 황정의 면역조절 효과 비교)

  • Park, Si-Deok;Lee, Geum-Hong;Lee, Young-Sun;Kwon, Young-Kyu;Park, Jong-Hyun;Choi, Sun-Mi;Shin, Sang-Woo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.2
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    • pp.414-424
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    • 2007
  • Adenophorae Radix (AR), Liriopis Tuber (LT), Dendrobii Monile (DM), Polygonati Officinalis (PO), Polygonati Rhizoma (PR) have been used to treated a variety condition/diseases in traditional oriental medicine. The present study was conducted to investigate the immunmodulatory effects of the water-extracted AR, LT, DH, PO and PR. In spleen cell proliferation assay, DH was significantly enhanced mitogenic activity compared with control group. In RT-PCR, DH ad PO induced IL-2 and IFNr cytokine gene expression in mouse spleen cells. Methotrexate(MTX), immune supression agent, was significantly inhibited mouse spleen cell proliferation(1600 mg/ml). In spite of MTX treatement, DH and PO sustained the spleen cell proliferation, In the flow cytometry analysis, DH stimulated mouse spleen cells showed an increase in B-cell phenotype (CD45R/B220). The water-extracted DH and PO inhibited NO production and iNOS expression in LPS-stimulated RAW 264.7 macrophage cell. DH induced IL-2 and IFNg gene expression in human peripheral mononuclear cells. The GC-MS analysis show that the main component of water-extracted DH was b-Nitroethyl alcohol. The main components of water-extracted PO were Dipirartril-tropico, Methyl sulfoxide and Demsodrox. These data suggest that among these extracts, DH has a protective effcet of immune suppression caused by MTX. DH may be enhance cellular and humoral immune response by the regulation of cytokine gene expression, NO production and B cell production.

EFFECTS OF LENTINUS EDODES LECTINS ON CYTOKINE GENE EXPRESSION FROM HUMAN PERIPHERAL BLOOD MONONUCLEAR CELLS (표고버섯 렉틴의 사이토카인 생성 양상 및 특성)

  • 이인경;김희선;전경희;김성광;정시련
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.113-113
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    • 1995
  • 본 연구는 표고버섯에서 분리정제한 렉틴 성분(LEL)을 말초혈액 단핵세포(PBMC)에 반응시켜 사이토카인 유도능을 지질다당류(LPS)와 비교하여 역전사효소 중합반응법(RT-PCR)으로 측정하였다. 측정 대상 사이토카인은 IL-1, IL-2, IL-6, TNF$\alpha$ 및 IFN${\gamma}$의 다섯가지였으며 이들을 대상으로 PBMC에 렉틴을 적용하여 1, 8, 24, 48, 72, 96, 120분 등의 시간대에서 반응시켜 사이토카인의 유전자 발현 유도에 관한 다음의 결과를 얻었다. LEL의 사용 농도에 따른 편도선 림프구(tonsillar lymphocyte)의 TNF$\alpha$ 유전자 발현 양상은 반응 1시간의 경우 LEL의 일부 농도와 LPS 전농도에서 관찰되었으나 반응 40시간째에는 LEL 전농도와 LPS 전농도에서 TNF$\alpha$ 유전자 발현 양상을 관찰할 수 없었다. RT-PCR 결과 원액이나 회석액 재료로부터 관찰된 TNF$\alpha$유전자 band의 강 약 차이는 나타나지 않았다. LEL의 자극에 의한 반응 시간대 별 PBMC에 의한 사이토카인 유전자 발현 양상은 위에서 언급된 다섯가지 사이토카인을 유도, 생성할 수 있다는 것이 확인되었는데, IL-2, IL-6 및 IFN${\gamma}$는 120시간까지 장시간 지속되는 유전자 발현이 가능한 반면 TNF $\alpha$의 생성 양상은 이들 사이토카인의 생성 양상과는 판이하게 반응 1, 8 및 24 시간대까지만 TNF$\alpha$ 유전자 발현을 관찰할 수 있었고 IL-1은 72 시간까지 반응을 나타내는 등 특이적 양상을 보였다. 한편 LPS는 실험에 사용된 전 사이토카인의 유전자 발현을 120시간대까지 반응이 유지됨을 관찰하였기에 LPS가 PBMC의 강력한 사이토카인 유도체임을 입증할 수 있었으며 LEL과 다소 상이한 결과를 보였으나 LEL 또한 PBMC로부터 사이토카인을 생성 유지시킬 수 있는 유도체로 작용함을 확인할 수 있었다.

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Effect of PHA and conditioned medium on blastogenesis and rosette formation of bovine circulating blood lymphocytes (PHA 및 conditioned medium 이 소의 순환혈액 림프구의 유약화와 rosette 형성에 미치는 영향)

  • Kang, Sei-woong;Yoon, Chang-yong;Song, Hee-jong
    • Korean Journal of Veterinary Research
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    • v.34 no.2
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    • pp.301-306
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    • 1994
  • This study was planned to estimate the activity of bovine circulating blood lymphocytes using phytohemagglutinin-M(PHA) known as T cell mitogen. Bovine circulating blood mononuclear cells(MNCs) was separated, and cultured with or without macrophage($PHA^+/M{\phi}^+$ or $PHA^+/M{\phi}^-$) in conditioned medium which stimulated with various concentration of PHA(0, 5, 10, 15 and $20{\mu}g/ml$ in medium), and then investigated the blastogenic response and rosette formation of lymphocytes. Blastogenic rate(BR) was especially increased in PHA concentration(10 and $15{\mu}g/ml$) of $PHA^+/M{\phi}^+$ group and their BR were $41.5{\pm}6.8%$ and $44.4{\pm}8.9%$, respectively and BR in PHA concentration(15 and $20{\mu}g/ml$) of $PHA^+/M{\phi}^-$ group was $32.8{\pm}6.2%$ and $31.4{\pm}4.6%$, respectively. BR of lymphocytes was more increased in $PHA^+/M{\phi}^+$ than $PHA^+/M{\phi}^-$ group when these cells were stimulated by PHA. Rosette forming rate(RFR) of lymphocytes to SRBC highly increased when SRBC was treated with AET and/or dextran, respectively. On the orther hand, RFR significantly increased more in $PHA^+/M{\phi}^+$ and $PHA^+/M{\phi}^-$ group than in control group, but when compared with two groups, statistical significancy was recognized only in PHA concentration($15{\mu}g/ml$, p<0.026) of $PHA^+/M{\phi}^+$ group.

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Preliminary assessment of correlation between T-lymphocyte responses and control of porcine reproductive and respiratory syndrome virus (PRRSV) in piglets born after in-utero infection of a type 2 PRRSV

  • Cha, Sang-Ho;Bandaranayaka-Mudiyanselage, Carey;Bandaranayaka-Mudiyanselage, Chandima B.;Ajiththos, Dharani;Yoon, Kyoung-Jin;Gibson, Kathleen A.;Yu, Ji-Eun;Cho, In-Soo;Lee, Stephen S.;Chung, Chungwon J.
    • Korean Journal of Veterinary Research
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    • v.58 no.1
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    • pp.9-16
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    • 2018
  • A preliminary study into the protective mechanisms of adaptive immunity against porcine reproductive and respiratory syndrome virus (PRRSV) in piglets (n = 9) born to a gilt challenged intranasally with a type-2 PRRSV. Immune parameters (neutralizing antibodies, $CD3^+CD4^+$, $CD3^+CD8^+$, $CD3^+CD4^+CD8^+$ T-lymphocytes, and PRRSV-specific interferon $(IFN)-{\gamma}$ secreting T-lymphocytes) were compared with infection parameters (macro- and microscopic lung lesion, and PRRSV-infected porcine alveolar macrophages ($CD172{\alpha}^+PRRSV-N^+\;PAM$) as well as with plasma and lymphoid tissue viral loads. Percentages of three T-lymphocyte phenotypes in 14-days post-birth (dpb) peripheral blood mononuclear cell (PBMC) had significant negative correlations with percentages of $CD172{\alpha}^+PRRSV-N^+\;PAM$ (p < 0.05) as well as with macroscopic lung lesion (p < 0.01). Plasma and tissue viral loads had significant (p < 0.05) negative correlations with $CD3^+CD4^+CD8^+$ T-lymphocyte percentage in PBMC. Frequencies of $CD3^+CD8^+$ and $CD3^+CD4^+$ T-lymphocytes in 14-dpb PBMC had significant negative correlations with of lymph node (p = 0.04) and lung (p = 0.002) viral loads. $IFN-{\gamma}$-secreting T-lymphocytes frequency had a significant negative correlation with gross lung lesion severity (p = 0.002). However, neutralizing antibody titers had no significant negative correlation (p > 0.1) with infection parameters. The results indicate that T-lymphocytes contribute to controlling PRRSV replication in young piglets born after in-utero infection.

In vitro cytotoxicity of four calcium silicate-based endodontic cements on human monocytes, a colorimetric MTT assay

  • Khedmat, Sedigheh;Dehghan, Somayyeh;Hadjati, Jamshid;Masoumi, Farimah;Nekoofar, Mohammad Hossein;Dummer, Paul Michael Howell
    • Restorative Dentistry and Endodontics
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    • v.39 no.3
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    • pp.149-154
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    • 2014
  • Objectives: This study was performed to evaluate the cytotoxicity of four calcium silicate-based endodontic cements at different storage times after mixing. Materials and Methods: Capillary tubes were filled with Biodentine (Septodont), Calcium Enriched Mixture (CEM cement, BioniqueDent), Tech Biosealer Endo (Tech Biosealer) and ProRoot MTA (Dentsply Tulsa Dental). Empty tubes and tubes containing Dycal were used as negative and positive control groups respectively. Filled capillary tubes were kept in 0.2 mL microtubes and incubated at $37^{\circ}C$. Each material was divided into 3 groups for testing at intervals of 24 hr, 7 day and 28 day after mixing. Human monocytes were isolated from peripheral blood mononuclear cells and cocultered with 24 hr, 7 day and 28 day samples of different materials for 24 and 48 hr. Cell viability was evaluated using an MTT assay. Results: In all groups, the viability of monocytes significantly improved with increasing storage time regardless of the incubation time (p < 0.001). After 24 hr of incubation, there was no significant difference between the materials regarding monocyte viability. However, at 48 hr of incubation, ProRoot MTA and Biodentine were less cytotoxic than CEM cement and Biosealer (p < 0.01). Conclusions: Biodentine and ProRoot MTA had similar biocompatibility. Mixing ProRoot MTA with PBS in place of distilled water had no effect on its biocompatibility. Biosealer and CEM cement after 48 hr of incubation were significantly more cytotoxic to on monocyte cells compared to ProRoot MTA and Biodentine.