• Title/Summary/Keyword: Interferon-gamma assay

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Effects of IFN-γ on IL-18 Expression in Pregnant Rats and Pregnancy Outcomes

  • Si, Li-Fang;Zhang, Shou-Yan;Gao, Chun-Sheng;Chen, Shu-Lin;Zhao, Jin;Cheng, Xiang-Chao
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.10
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    • pp.1399-1405
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    • 2013
  • The present study focused on establishing the effects of interferon-gamma (IFN-${\gamma}$) on interleukin-18 (IL-18) expression patterns and pregnancy outcomes in pregnant rats. Pregnant rats at the post-implantation stage were randomized into control, low IFN-${\gamma}$ (L-IFN-${\gamma}$) and high IFN-${\gamma}$ groups (H-IFN-${\gamma}$) that received normal saline, 100 IU/g of IFN-${\gamma}$ and 500 IU/g of IFN-${\gamma}$ vaginal muscular injection, respectively. The effects of IFN-${\gamma}$ on IL-18 expression and pregnancy outcomes were assessed systematically using several methods, including immunohistochemistry streptavidin-perosidase (SP), image pattern analysis, enzyme-linked immune-sorbent assay (ELISA), whole blood count (WBC) count, microscopy and visual observation. IL-18 was detected in the uteri of all pregnant rats, and mainly distributed in the endometrium, decidual cells, vascular endothelium and myometrium. Immunohistochemistry and image pattern analyses revealed significantly lower IL-18 expression in the H-IFN-${\gamma}$ group compared to the L-IFN-${\gamma}$ and control groups (p<0.01), indicating that high doses of IFN-${\gamma}$ induce downregulation of IL-18 in the uterus of pregnant rats. ELISA results disclosed that IL-18 expression in peripheral blood of the H-IFN-${\gamma}$ group was lower than that of the L-IFN-${\gamma}$ group (p<0.05), and significantly reduced compared to the control group (p<0.01). Moreover, the number of peripheral leukocytes in the H-IFN-${\gamma}$ group was significantly higher than those in the control and L-IFN-${\gamma}$ groups (p<0.01). Morphology analysis showed no evident differences between the L-IFN-${\gamma}$ and control groups. However, for the H-IFN-${\gamma}$ group, uterine mucosa bleeding, necrosis and excoriation were observed using microscopy. Visual observation revealed marroon, swelling, crassitude and no embryo in the uterus, which are obvious indicators of abortion. These results indicate that IFN-${\gamma}$ plays a regulatory role in IL-18 expression in the uterus and peripheral blood of pregnant rats at the post-implantation stage. Moreover, high levels (500 IU/g) of IFN-${\gamma}$ influence normal pregnancy at the early stages in rats by downregulating IL-18 expression in the uterus and peripheral blood and increasing the number of peripheral leukocytes, consequently triggering termination of pregnancy.

Diagnosis for Latent Tuberculosis Infection in College Students (대학생 잠복결핵 감염의 진단)

  • Yook, Keun-Dol;Yang, Byoung-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.225-229
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    • 2016
  • Tuberculosis (TB) is caused by a chronic infectious agent known as Mycobacterium tuberculosis. It is transmitted in airborne particles, called droplet nuclei which was generated by cough, sneeze, shout, or sing of persons who have TB disease. Most infections of TB do not have symptoms, well known as latent tuberculosis infection (LTBI). However, about 10% of LTBI progress to active disease a one or two years after infection. To investigate the LTBI rate of college students who were in contacted with TB patients, we performed chest X-ray, tuberculin skin test (TST) and Interferon-gamma release assay (IGRA) to 74 college students. At a results, 65 students were showed negative and 9 students positive results at chest X-ray and 1st TST test. When confirmed the 65 students who were showed negative by 2st TST, the results showed correctly. But, 9 students who were showed positive results on chest X-ray and 1st TST by IGRA, the only 3 students (4.05%) showed positive results. In conclusion, the LTBI rate in this study showed 4.05% (3/74) and we suggest to investigate other students LTBI rate for decreasing tuberculosis.

A Tapping the usefulness of Whole Blood Interferon-${\gamma}$ Assay for Diagnosing Tuberculosis Infection in Children (소아 결핵 감염 진단에 있어서 결핵 특이항원 자극 Interferon-${\gamma}$ 분비능 측정의 진단적 유용성)

  • Soon, Eu-Gene;Lim, Baek-Keun;Kim, Hwang-Min;NamGoong, Mee-Kyung;Cha, Byung-Ho;Uh, Young;Chun, Jin-Kyong
    • Tuberculosis and Respiratory Diseases
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    • v.68 no.5
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    • pp.280-285
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    • 2010
  • Background: $QuantiFERON^{(R)}$-TB Gold In Tube (QFT-G IT) has been used for diagnosing latent tuberculosis infection and active tuberculosis (TB) since 2007. However, there has not been enough data on QFT-G IT for universal use in children. In this study, we evaluated the clinical usefulness of the QFT-G IT in pediatric practice. Methods: We retrospectively reviewed the clinical records of 70 patients younger than 18 years of age who had taken QFT-G IT and had a tuberculin skin test (TST) between July 2007 and July 2009 at Wonju Christian Hospital. The subjects were divided into two groups, asymptomatic TB exposure group and disease group. Four patients who were taking immunosuppressants during the study period were excluded. Results: A total of 66 immunocompetent children were included in this study. Among 27 asymptomatic children who had contact histories of TB, 6 (22.2%) were found to be positive by QFT-G IT. Eleven (40.7%) and 5 (18.5%) children were found to be positive by TST with cutoff values of ${\geq}5mm$ and ${\geq}10mm$, respectively. Agreement was fair to good between QFT-G IT and TST (${\kappa}=0.59$: cutoff value ${\geq}5mm$, ${\kappa}=0.7$: cutoff value ${\geq}10mm$). In disease group, 14 patients (35.9%) were diagnosed with active tuberculosis, 8/14 (57.1%) were positive on TST and 9/14 (64.3%) on QFT-G IT. The positive rate of acid-fast bacilli smear, TB-polymerase chain reaction, and culture for tuberculosis was 11% (1/9), 27.3% (3/11) and 33.3% (3/9), respectively. Conclusion: Our data support that the QFT-G IT can be used as an additional diagnostic tool for latent and active tuberculosis infection in children.

Pregnancy influences expression of interferon-stimulated genes, progesterone receptor and progesterone-induced blocking factor in ovine thyroid

  • Jianhua Cao;Shuxin Zhao;Yaqi Zhang;Jiabao Cai;Leying Zhang;Ling Yang
    • Animal Bioscience
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    • v.37 no.8
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    • pp.1377-1386
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    • 2024
  • Objective: Embryonic interferon-tau (IFNT) and progesterone affect expression of interferon-stimulated genes (ISGs), progesterone receptor (PGR) and progesterone-induced blocking factor (PIBF) in the ovine thyroid. Methods: Thyroids of ewes were sampled at day 16 of nonpregnancy, days 13, 16, and 25 of pregnancy, and real-time quantitative polymerase chain reaction assay, western blot and immunohistochemistry were used to detect expression of ISGs, PGR, and PIBF. Results: Free ISG15 protein was undetected, but ISG15 conjugated proteins upregulated at day 16 of pregnancy, and expression levels of ISG15 conjugated proteins, PGR isoform (70 kDa), PIBF, interferon-gamma-inducible protein 10 and myxovirusresistance protein 1 peaked, but expression level of signal transducer and activator of transcription 1 was the lowest at day 16 of pregnancy. In addition, the expression levels of PGR isoform (70 kDa) and signal transducer and activator of transcription 1 (STAT1) decreased, but levels of PGR isoform (43 kDa), 2',5'-oligoadenylate synthetase, IP-10 and MX1 increased at day 25 of pregnancy comparing with day 16 of the estrous cycle. Conclusion: Early pregnancy affects expression of ISGs, PGR, and PIBF in maternal thyroid through IFNT and progesterone, which may regulate thyroid autoimmunity and thyroid hormone secretion in ewes.

Immunomodulating and Anticoagulant Activity of Glycosaminoglycans Derived from Porcine Testis

  • Yoo, Yung-Choon;Kim, Yeong-Shik;Song, Kyung-Sik;Moon, Eun-Ho;Lee, Kyung-Bok
    • Archives of Pharmacal Research
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    • v.25 no.5
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    • pp.669-674
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    • 2002
  • Glycosaminoglycans (GAGs) were isolated from the porcine testis, and their immuno-modulating and anticoagulant activity was investigated. From anion exchange chromatography (Dowex Macropolous Resin) used for further isolation of porcine testis GAGs (PT-GAGs), two fractions (PT-GAG-1.5 and PT-GAG-16) eluted by different salt concentration were obtained. In immunomodulating activity test, PT-GAG-1.5, but not PT-GAG-16, significantly enhanced the growth of murine peritoneal macrophages. In addition, treatment with PT-GAG-1.5 induced the production of cytokines, interleukin-1$\beta$ (IL-1$\beta$), interferon-${\gamma}$ (IFN-${\gamma}$) and tumor necrosis factor-$\alpha$ (TNF-$\alpha$), from murine microphages. Unexpectedly, both of PT-GAGs had no effect on the growth of murine splenocytes. The anticoagulant activity of PT-GAG-1.5 and PT-GAG-16 was examined by activated partial thromboplastin time (aPTT) assay and thrombin time (TT) assay. Both of PT-kGAGs significantly increased the clotting times of aPTT and TT in a dose-dependent manner. The anticoagulant activity of PT-GAG-16 was found to be higher than that of PT-GAG-1.5. These results suggest that PT-GAGs possess biological activities such as immunomodulating activity and anticoagulant activity.

Upregulation of IP-10(CXCL10) mRNA Expression by Interleukin-18

  • Kim, Hyo-Young;Kim, Hee-Sun
    • Journal of Yeungnam Medical Science
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    • v.24 no.1
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    • pp.67-78
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    • 2007
  • Background : Interleukin-18 (IL-18) is one of the principal inducers of interferon-${\gamma}$ (IFN-${\gamma}$) in lymphocytes. Materials and Methods : The effect of IL-18 on the expression of chemokine IP-10(CXCL10) mRNA in C57BL/6 mouse peritoneal macrophages was studied by using Northern blot analysis, enzyme linked immunosobent assay and electrophoretic mobility shift assay. Results : IL-18 was determined to exert no direct effect on the expression of IP-10(CXCL10) mRNA. However, IL-18 pretreatment was determined to play a cooperative role in the synergistic induction of LPS-induced IP-10(CXCL10) mRNA expression. The effect associated with IL-18 pretreatment with regard to the synergistic induction of LPS-induced IP-10 (CXCL10) mRNA expression was detected after 16 hr of IL-18 pretreatment, administered prior to LPS stimulation. The pattern of NF-${\kappa}B$ binding activity during IL-18 pretreatment with LPS stimulation was found to coincide with the expression of IP-10(CXCL10) mRNA. Conclusion : Although IL-18 alone exerts no direct effect on the expression of chemokine IP-10(CXCL10), a definite period of IL-18 pretreatment induces the synergistic expression of LPS-induced IP-10(CXCL10) mRNA. NF-${\kappa}B$ activation is a component of this synergistic effect of IL-18 pretreatment. These results provide useful information, which may facilitate the elucidation of the action mechanisms underlying IL-18 effect on the expression of IP-10(CXCL10) mRNA.

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Properties of the Endonuclease Secreted by Human B Lymphoblastic IM9 Cells

  • Kwon, Hyung-Joo;Kim, Doo-Sik
    • BMB Reports
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    • v.31 no.1
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    • pp.106-110
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    • 1998
  • We have employed a DNA-native-polyacrylamide gel electrophoresis (DNA-native-PAGE) assay system to characterize the enzyme activity of the endonuclease secreted by human B lymphoblastic IM9 cells. Experimental results clearly demonstrated that the endonuclease activity of IM9 cell culture medium is distinct from that of DNase I in the DNA-native-PAGE assay system. Immunoprecipitation analysis using anti-DNase I antibody showed that the secreted endonuclease is not recognized by the antibody. The secreted endonuclease was estimated using supercoiled plasmid DNA as a substrate. The pH optimum required for the catalytic activity was determined to be in the range of pH 6.6-7.4. No significant difference in the endonuclease secretion was observed by stimulation of the IM9 cells with interferon-${\gamma}$ or interleukin-$1{\beta}$.

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Atopic Dermatitis-Related Inflammation in Macrophages and Keratinocytes: The Inhibitory Effects of Bee Venom

  • Kim, Deok-Hyun;Song, Ho-Sueb
    • Journal of Acupuncture Research
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    • v.36 no.2
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    • pp.80-87
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    • 2019
  • Background: This study investigated the anti-inflammatory effects of bee venom (BV) through the inhibition of nuclear factor kappa beta ($NF-{\kappa}B$) expression in macrophages and keratinocytes. Methods: Cell viability assays were performed to investigate the cytotoxicity of BV in activated macrophages [lipopolysaccharide (LPS)] and keratinocytes [interferon-gamma/tumor necrosis factor-alpha ($IFN-{\gamma}/TNF-{\alpha}$)]. A luciferase assay was performed to investigate the cellular expression of $NF-{\kappa}B$ in relation to BV dose. The expression of $NF-{\kappa}B$ inhibitors ($p-I{\kappa}B{\alpha}$, $I{\kappa}B{\alpha}$, and p50 and p65) were determined by Western Blot analysis, and the electromobility shift assay. A nitrite quantification assay was performed to investigate the effect of BV, and $NF-{\kappa}B$ inhibitor on nitric oxide (NO) production in macrophages. In addition, Western Blot analysis was performed to investigate the effect of BV on the expression of mitogen-activated protein kinases (MAPK) in activated macrophages and keratinocytes. Results: BV was not cytotoxic to activated macrophages and keratinocytes. Transcriptional activity of $NF-{\kappa}B$, and p50, p65, and $p-I{\kappa}B{\alpha}$ expression was reduced by treatment with BV in activated macrophages and keratinocytes. Treatment with BV and an $NF-{\kappa}B$ inhibitor, reduced the production of NO by activated macrophages, and also reduced $NF-{\kappa}B$ transcriptional activity in activated keratinocytes (compared with either BV, or $NF-{\kappa}B$ inhibitor treatment). Furthermore, BV decreased p38, p-p38, JNK, and p-JNK expression in LPS-activated macrophages and $IFN-{\gamma}/TNF-{\alpha}$-activated keratinocytes. Conclusion: BV blocked the signaling pathway of $NF-{\kappa}B$, which plays an important role in the inflammatory response in macrophages and keratinocytes. These findings provided the possibility of BV in the treatment of atopic dermatitis.

Anti-Inflammatory Effect of Ligustri Lucidi Fructus Water Extract in RAW 264.7 Cells Induced by LPS (여정실(女貞實)이 LPS로 유발된 RAW 264.7 cell의 염증에 미치는 영향)

  • Lee, Yong-Hyun;Lim, Eun-Mee
    • The Journal of Korean Obstetrics and Gynecology
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    • v.26 no.4
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    • pp.66-81
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    • 2013
  • Purpose: This study was carried out to investigate the anti-inflammatory effects of Ligustri Lucidi Fructus water extract (LF) in the lipopolysaccharide (LPS)-induced mouse macrophages RAW 264.7 cell. Methods: Ligustri Lucidi Fructus was extracted with distilled water (2,000 ml) for 2 hours. In order to evaluate cytotoxicity of LF, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed. To investigate anti-inflammatory effects of LF, the concentration of nitric oxide (NO) was measured with NO assay, cytokine was measured by Bio-Plex cytokine assay, and intracellular calcium (Ca) was measured with Fluo-4 Ca assay in RAW 264.7 cell. And when p-value is below 0.05, it is judged to have the significant difference statistically (P<0.05). Results: 1. LF showed no cytotoxicity. 2. LF inhibited significantly the production of NO at the concentration of 25, 50 and $100{\mu}g/ml$. 3. LF inhibited significantly the production of interleukin (IL)-4, macrophage inflammatory protein (MIP)-$1{\alpha}$, granulocyte colony stimulating factor (G-CSF) at the concentration of 25, 50, 100 and $200{\mu}g/ml$. 4. LF inhibited significantly the production of granulocyte macrophage-colony stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF) at the concentration of 50, 100 and $200{\mu}g/ml$, the interferon (IFN)-${\gamma}$ at 25, 50 and $100{\mu}g/ml$ respectively. 5. LF inhibited significantly the production of IL-$1{\beta}$ at the concentration of 50 and $200{\mu}g/ml$, the IL-5 at 25 and $100{\mu}g/ml$, the IL-12p70, MIP-$1{\beta}$ at 50 and $100{\mu}g/ml$, the regulated on activation, normal T cell expressed and secrete d (RANTES) at 100 and $200{\mu}g/ml$ respectively. 6. LF inhibited significantly the production of IL-10, interferon gamma-induced protein (IP)-10 at the concentration of $200{\mu}g/ml$. 7. LF inhibited significantly the production of intracellular Ca at the concentration of 25, 50, 100 and $200{\mu}g/ml$. Conclusions: These results suggest that LF has anti-inflammatory effect and immuno-modulating activity.

Studies on the Regulatory Effect of Cytokine Production in Taumin Patients with Cerebral Infarction by Cheongsimyeonjatang (청심연자탕(淸心蓮子湯)이 태음인(太陰人) 뇌경색증(腦硬塞症) 환자(患者)의 세포활성물질(細胞活性物質) 생성조절(生成調節)에 미치는 영향(影響))

  • Kim, Kyung-Yo;Noh, Hyun-Soo
    • Journal of Sasang Constitutional Medicine
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    • v.12 no.2
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    • pp.162-170
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    • 2000
  • Purpose : Studies on the Regulatory Effect of Cytokine Production in Taumin Patients with Cerebral Infarction by Cheongsimyeonjatang Method : ELISA(enzyme-linked immuno-sorbent assay) Result : Chungsimyeunjatang(CYT) is a prescription for the cerebral infarction (CI) patients of Taeumin according to Sasang constitution philosophy. Taeumin patients with CI were treated with CYT during the acute stage. Clinical signs of CI disappeared markedly in about two to four weeks after oral administration of CYT in all patients. The mean interleukin (IL)-2 plasma levels were slightly lower in the patients with CI than in the normal groups, whereas the mean IL-4, IL-6 and IgE levels were significantly higher in the patients. There were no significant differences in $interferon-{\gamma}$ $(IFN-{\gamma})$ levels between the groups. Serum $IFN-{\gamma}$ and IL-2 levels derived from T helper (Th)1 cells were elevated significantly in the patients with CI by CYT administration. Significant reduced plasma levels of IL-4 and IL-6 derived from Th2 cells and IgE were observed in the patients treated with CYT. During the period of CYT administration, there were no other adverse effects. The data indicate that CYT has a good CI treatment effect, and that its action may be due to regulation of cytokine Production.

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