• Title, Summary, Keyword: $interferon{\gamma}$

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Specific Expression of Interferon-γ Induced by Synergistic Activation Mediator-Derived Systems Activates Innate Immunity and Inhibits Tumorigenesis

  • Liu, Shuai;Yu, Xiao;Wang, Qiankun;Liu, Zhepeng;Xiao, Qiaoqiao;Hou, Panpan;Hu, Ying;Hou, Wei;Yang, Zhanqiu;Guo, Deyin;Chen, Shuliang
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1855-1866
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    • 2017
  • The synergistic activation mediator (SAM) system can robustly activate endogenous gene expression by a single-guide RNA. This transcriptional modulation has been shown to enhance gene promoter activity and leads to epigenetic changes. Human $interferon-{\gamma}$ is a common natural glycoprotein involved in antiviral effects and inhibition of cancer cell growth. Large quantities of high-purity $interferon-{\gamma}$ are important for medical research and clinical therapy. To investigate the possibility of employing the SAM system to enhance endogenous human $interferon-{\gamma}$ with normal function in innate immunity, we designed 10 single-guide RNAs that target 200 bp upstream of the transcription start sites of the $interferon-{\gamma}$ genome, which could significantly activate the $interferon-{\gamma}$ promoter reporter. We confirmed that the system can effectively and highly activate $interferon-{\gamma}$ expression in several humanized cell lines. Moreover, we found that the $interferon-{\gamma}$ induced by the SAM system could inhibit tumorigenesis. Taken together, our results reveal that the SAM system can modulate epigenetic traits of non-immune cells through activating $interferon-{\gamma}$ expression and triggering JAK-STAT signaling pathways. Thus, this strategy could offer a novel approach to inhibit tumorigenesis without using exogenous $interferon-{\gamma}$.

EFFECTS OF $INTERFERON-\gamma$ ON COLLAGEN AND FIBRONECTIN SYNTHESIS IN PRIMARY CULTURED PERIODONTAL LIGAMENT CELLS ($Interferon-\gamma$가 치주인대 세포의 Collagen 및 Fibronectin의 합성과 Alkaline Phosphatase 활성에 미치는 영향)

  • Kim, Gwang-Seok;Sung, Jae-Hyun;Choi, Je-Yong;Ryou, Hyun-Mo
    • The korean journal of orthodontics
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    • v.23 no.2
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    • pp.229-248
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    • 1993
  • [ $Interferon-\gamma$ ] has been suggested as a cytokine of connective tissue stabilizer. In addition, it has also been demonstrated that this cytokine inhibited bone remodeling activities of the bone derived cells. In order to illuminate the effects of this cytokine in orthodontic force induced bone remodeling, it was administered to primary cultured periodontal ligament cells which have been known to have some osteoblast like characteristics. $Interferon-\gamma$ slightly decreased $[^3H]thymidine$ incorporation rate without a significant change in the total cellular DNA content up to 1000 U/ml, which meant these doses were not cytotoxic to the cell. Total protein synthesis was not influenced by various concentration of interferon-y whether it was determined by the $[^3H]proline$ incorporation rate or by the Lowry smethod. The effect of $interferon-\gamma$ on the individual protein was, however, differential, ie, it increased $[^3H]proline$ incorporation into the noncollagenous protein marginally, while it decreased $[^3H]proline$ incorporation into the collagen, so that it caused dose-dependent suppression of the relative collagen synthesis. On the contrary, the fibronectin synthesis determined by the ELISA was increased by 1000 U/ml of $interferon-\gamma$. The differential effects of the interferon-y on the collagen and fibronectin synthesis exhibited not only their protein level but also the steady state mRNA level. $Interferon-\gamma$ decreased steady state level of ${\alpha}1(I)$ procollagen mRNA significantly, while showing no significant changes in the fibronectin mRNA level. In addition to this, it was also found that indomethacin did not affect on the $interferon-\gamma$ induced collagen decrease in this cell, which meant prostaglandins were not involed in the process of $interferon-\gamma$ induced collagen decrease. So it can be concluded that the incubation of periodontal ligament cells with 1000 U/ml of $interferon-\gamma$ for 24 hr showed differential effects on the type I collagen and fibronectin gene expression. The decrease in relative collagen synthesis in the protein level was related with decrease in the steady state level of mRNA, while the increase in the fibronectin synthesis in the protein level was not correlated with the mRNA level.

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The Mechanism of Interferon-$\gamma$ Induced Cytotoxicity on the Lung Cancer Cell Line, A549 (인터페론감마에 의한 A549 폐암세포주 세포독성의 기전)

  • Oh, Yeon-Mok;Yoo, Chul-Gyu;Chung, Hee-Soon;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.1
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    • pp.63-68
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    • 1996
  • Background: Interferon-$\gamma$ has various biologic effects, including antiviral effect, antitumor proliferative effect, activation of macrophage and B lymphocyte, and increased expression of major histocompatibility complex. Especially, antitumor proliferative effect of interferon-$\gamma$ has already been proved to be important in vivo as well as in vitro. And, clinical studies of interferon-$\gamma$ have been tried in lung cancer patients. However, the mechanism of antitumor effect of interferon-$\gamma$ has not yet been established despite of many hypotheses. "Necrosis" is a type of cell death which is well known to occur in the circumstances of severe stresses. In contrast, "apoptosis" is another type of cell death which occurs in such biological circumstances as embryonic development, regression of organs, and self-tolerance of lymphocytes. And, apoptosis is an active process of cell death in which cells are dying with fragmentations of their cytoplasms and nuclei. And, in the process of apoptosis the DNAs of cells are cleaved between nucleosomes by unidentified endonuclease and therefore DNAs of apoptotic cells result in a typical electrophoresis pattern known as DNA ladder pattern. Recently it has been suggested that cytotoxic effect of interferon-$\gamma$ occurs via apoptosis. To elucidate the mechanism of antitumor cytotoxic effect of interferon-$\gamma$, we microscopically observed a lung cancer cell line, A549 which was treated with interferon-$\gamma$. We observed A545 treated with interferon-$\gamma$ was dying fragmented. And so, we performed this study to find out that the mechanism of antitumor cytotoxic effect of interferon-$\gamma$ be apoptosis. Method: We treated A549, human lung cancer cell line with various concentration of interferon-$\gamma$ and quantified its cytotoxic effect of various periods, 24 hours, 72 hours and, 120 hours by MTT(dimethylthiazolyl diphenyltetrazolium bromide) bioassay. Also, after we treated A549 with 100 units/mi of interferon-$\gamma$ for 120 hours, we observed the pattern of cell death with inverted microscope and we extracted DNAs from the dead A549 cells and observed the pattern of 1.5% agarose gel electrophoresis with ethidium bromide staining. Result: 1) Cytotoxic effect of interferon-$\gamma$ on A549: For the first 24 hours, threre was little cytotoxic effect and for between 24 hours and 72 hours, there was the beginning of cytotoxic effect and for 120 hours there was increased cytotoxic effect. 2) Pattern of A549 cell death by interferon-$\gamma$: We observed with inverted microscope that A549 cells were dying fragmented. 3) DNA ladder pattern of gel electrophoresis: We observed DNA ladder pattern of gel electrophoresis of extracted DNAs from dead A549 cells. Conclusion: We concluded that the mechanism of interferon-$\gamma$induced cytotoxicity on lung cancer cell line, A549 be via apoptosis.

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MOLECULAR CLONING OF CHICKEN INTERFERON-GAMMA (닭 인터페론 유전자의 클로닝에 관한 연구)

  • ;Hyun Lillehoj
    • Proceedings of the Korea Society of Poultry Science Conference
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    • pp.34-50
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    • 1999
  • A cDNA encoding chicken interferon-gamma (chIFN-${\gamma}$) was amplified from P34, a CD4$^{+}$ T-cell hybridoma by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into pUC18. THe sequences of cloned PCR products were determined to confirm the correct cloning. Using this cDNA as probe, chicken genomic library from White Leghorn spleen was screened. Phage clones harboring chicken interferon-gamma (chIFN-${\gamma}$) were isolated and their genomic structure elucidated. The chIFN-${\gamma}$ contains 4 exons and 3 introns spanning over 14 kb, and follows the GT/AG rule for correct splicing at the exon/intron boundaries. The four exons encode 41, 26, 57 and 40 amino acids, respectively, suggesting that the overall structure of IFN-${\gamma}$ is evolutionairly conserved in mammalian and avian species. The 5’-untranslated region and signal sequences are located in exon 1. Several AT-rich sequences located in the fourth exon may indicate a role in mRNA turnover. The 5’-flanking region contains sequences homologous to the potential binding sites for the mammalian transcription factors, activator protein-1(AP-1) activator protein-2(AP-2) cAMP-response element binding protein(CREB), activating transcription factor(ATF), GATA-binding fator(GATA), upstream stimulating factor(USF), This suggests that the mechanisms underlying transcriptional regulation of chicken and mammalian IFN-${\gamma}$ genes may be similar.r.

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Diagnosis of Mycobacterium tuberculosis Infection using Ex-vivo interferon-gamma Assay (체외 Interferon-gamma 검사를 이용한 결핵감염의 진단)

  • Lee, Jung Yeon;Shim, Tae Sun
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.5
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    • pp.497-509
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    • 2006
  • Until recently, the tuberculin skin test (TST) has been the only tool available for diagnosing a latent TB infection. However, the development of new diagnostic tools, using the Mycobacterium tuberculosis (MTB)-specific early secreted antigenic target 6 (ESAT-6) and culture filtrate protein 10 (CFP-10) antigens, should improve the control of tuberculosis (TB) by allowing a more accurate identification of a latent TB infection (LTBI). Antigen-specific interferon-gamma ($IFN-{\gamma}$) assays have greater specificity in BCG-vaccinated individuals, and as less biased by nontuberculous mycobacterial infections. Many comparative studies have suggested that those assays have a higher specificity than the TST, and the sensitivity of these assays are expected to remarkably improved if more MTB-specific antigens can become available. Nevertheless, the major obstacle to the widespread use of these tests is the limited financial resources. Similar to other diagnostic tests, the predictive value of $IFN-{\gamma}$ assays depends on the prevalence of a MTB infection in the population being tested. Therefore, prospective studies will be meeded to establish the applicability of these new assays at multiple geographic locations among patients of different ethnicities, and to determine if the $IFN-{\gamma}$ responses can indicate those with a high risk of progressing to active TB.

Inhibitory Effect on Adjuvant Arthritis and Other Pharmacological Profile of Gamma-Interferon, LBD-001 (감마 인터페론, LBD-001의 Adjuvant 관절염 억제작용 및 기타 일반 약리작용)

  • Lee, Eun-Bang;Kim, Jae-Hyun;Kim, Oon-Ja;Kim, Jung-Keun
    • YAKHAK HOEJI
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    • v.34 no.3
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    • pp.171-179
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    • 1990
  • The recombinant gamma-interferon (LBD-001) which was produced by yeast as host system was investigated on the pharmacological activities. This gamma-interferon exhibited potent inhibitory effect on adjuvant induced arthritis, but no effect on carrageenin induced paw edema in rats. It did not show any sedative, anticonvulsive, analgesic and hypothermic activities in animals. It also had no influences on isolated tracheal muscle and ileum of guinea pig, isolated uterus and fundus strip of rats, and on blood pressure and respiration in situ experiments of rabbits.

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The Inhibitory Activity of Erigeron annuus-Derived Components on $Interferon-{\gamma}$ and Lipopolysaccharide-Induced Nitric Oxide Production in Mouse Pheritoneal Macrophage

  • Lee, Hee-Jung;Kim, You-Ah;Jeong, Na-Ho;Hong, Seung-Heon;Seo, Young-Wan
    • Journal of Applied Biological Chemistry
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    • v.50 no.3
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    • pp.160-163
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    • 2007
  • Two flavonoids (1 and 2) and one phenolic acid (3) obtained from Erigeron annuus have recently been shown to have potent antioxidant activities. Aim of this study was to investigate the inhibitory effects of these components on $interferon-{\gamma}$ and lipopolysaccharide-induced nitric oxide productions in the mouse pheritoneal macrophage. Compounds 2 and 3 showed marked inhibitory activities against inducible nitric oxide synthase (iNOS) on the lipopolysaccharide and $interferon-{\gamma}-stimulated$ mouse pheritoneal macrophages without cytotoxicity. Therefore, these results suggest that the compounds could be effective anti-inflammatory agents as nitric oxide inhibitors in vivo.

Study on the therapeutic effects of interferon and gamma-globulin in experimental Pneumocustis curinii pneumonia (Interferon 및 gamma-globulin이 실험적 Pneumocystis carinii 폐염의 치료에 미치는 영향)

  • 신대환;강대영
    • The Korean Journal of Parasitology
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    • v.30 no.3
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    • pp.219-226
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    • 1992
  • This study was performed to observe the therapeutic effects of interferon-gamma ($IFN-{\gamma}$) and gamma-globulin (${\gamma}-globulin$) in experimental Pneumocystis carinii pneumonia of immune suppressed mice. After 9 weeks, trimethoprim-sulfamethoxaBole(TMP-SMZ; 10~50 mg/mouse/day), mouse $IFN-{\gamma}(5{\times}10^4$ units/mouse/day) and mouse ${\gamma}-globulin$(20 mg/mouse/day) were administered to the mice for 3 weeks by the experimental group. The therapeutic efficacy was evaluated by body weights, histopatholo단ic and electron microscopic findings of the lungs, and number of p. carinii cysts by Gomori's methenamine silver stain. Body weights of the mice were significantly increased in the group of combination therapy of TMP-SMZ with $IFN-{\gamma}{\;}or{\;}{\gamma}-globulin$, and in the group of TMP- SMZ treatment(p<0.05), however, little effect was found in the group of T-globulin alone. Histopathologic 6ndings of p. carinii pneumonia were much improved in the group of combination therapy of TMP-SMZ with $IFN-{\gamma}$. Treatment with either TMP-SMZ or $IFN-{\gamma}$ significantly reduced the number of cysts in the p. carinii pneumonia, but {\gamma}-globulin alone was ineffective. In electron microscopic findings of p. carinii pneumonia, the number of trophozoites and cysts were reduced by treatment with either TMP-SMZ or $IFN-{\gamma}$, and most of the cysts were empty or containing one or two intracystic bodies. The present results suggested, that combination therapy of TMP-SMZ with $IFN-{\gamma}$ had synergistic effects in treatment of P carinii pneumonia in experi- mental mice.

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The Poly-γ-ᴅ-Glutamic Acid Capsule of Bacillus licheniformis, a Surrogate of Bacillus anthracis Capsule Induces Interferon-Gamma Production in NK Cells through Interactions with Macrophages

  • Lee, Hae-Ri;Jeon, Jun Ho;Rhie, Gi-Eun
    • Journal of Microbiology and Biotechnology
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    • v.27 no.5
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    • pp.1032-1037
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    • 2017
  • The poly-${\gamma}$-$\small{D}$-glutamic acid (PGA) capsule, a major virulence factor of Bacillus anthracis, provides protection of the bacterium from phagocytosis and allows its unimpeded growth in the host. We investigated crosstalk between murine natural killer (NK) cells and macrophages stimulated with the PGA capsule of Bacillus licheniformis, a surrogate of the B. anthracis capsule. PGA induced interferon-gamma production from NK cells cultured with macrophages. This effect was dependent on macrophage-derived IL-12 and cell-cell contact interaction with macrophages through NK cell receptor NKG2D and its ligand RAE-1. The results showed that PGA could enhance NK cell activation by inducing IL-12 production in macrophages and a contact-dependent crosstalk with macrophages.

Usefulness of interferon-γ release assay for the diagnosis of latent tuberculosis infection in young children

  • Yun, Ki Wook;Kim, Young Kwang;Kim, Hae Ryun;Lee, Mi Kyung;Lim, In Seok
    • Korean Journal of Pediatrics
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    • v.59 no.6
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    • pp.256-261
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    • 2016
  • Purpose: Latent tuberculosis infection (LTBI) in young children may progress to severe active tuberculosis (TB) disease and serve as a reservoir for future transmission of TB disease. There are limited data on interferon-${\gamma}$ release assay (IGRA) performance in young children, which our research aims to address by investigating the usefulness of IGRA for the diagnosis of LTBI. Methods: We performed a tuberculin skin test (TST) and IGRA on children who were younger than 18 years and were admitted to Chung-Ang University Hospital during May 2011-June 2015. Blood samples for IGRA were collected, processed, and interpreted according to manufacturer protocol. Results: Among 149 children, 31 (20.8%) and 10 (6.7%) were diagnosed with LTBI and active pulmonary TB, respectively. In subjects lacking contact history with active TB patients, TST and IGRA results were positive in 41.4% (29 of 70) and 12.9% (9 of 70) subjects, respectively. The agreement (kappa) of TST and IGRA was 0.123. The control group, consisting of non-TB-infected subjects, showed no correlation between age and changes in interferon-${\gamma}$ concentration after nil antigen, TB-specific antigen, or mitogen stimulation in IGRAs (P=0.384, P=0.176, and P=0.077, respectively). In serial IGRAs, interferon-${\gamma}$ response to TB antigen increased in IGRA-positive LTBI subjects, but did not change considerably in initially IGRA-negative LTBI or control subjects. Conclusion: The lack of decrease in interferon-${\gamma}$ response in young children indicates that IGRA could be considered for this age group. Serial IGRA tests might accurately diagnose LTBI in children lacking contact history with active TB patients.