• Title/Summary/Keyword: T antigen

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Effects of Toxoplasma gondii and Toxocara canis Antigens on WEHI-164 Fibrosarcoma Growth in a Mouse Model

  • Darani, Hossein Yousofi;Shirzad, Hedayatollah;Mansoori, Fataneh;Zabardast, Nozhat;Mahmoodzadeh, Mahdi
    • Parasites, Hosts and Diseases
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    • v.47 no.2
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    • pp.175-177
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    • 2009
  • Cancer is the main cause of death in developed countries. However, in underdeveloped countries infections and parasitic diseases are the main causes of death. There are raising scientific evidences indicating that parasitic infections induce antitumor activity against certain types of cancers. In this study, the effects of Toxoplasma gondii and Toxocara canis egg antigens in comparison with Bacillus Calmette Guerin (BCG) (known to have anticancer distinctive) on WEHI-164 fibosarcoma transplanted to BALB/c mice was investigated. Groups of 6 male BALB/c mice injected with T. gondii antigen, BCG, or T. canis egg antigen as case groups and alum alone as control groups. All mice were then challenged with WEHI-164 fibrosarcoma cells. The mice were examined for growth of the solid tumor and the tumor sizes were measured every other day up to 4wk. The mean tumor area in T. gondii, BCG, or alum alone injected mice in 4 different days of measurements was $25\;mm^2$, $23\;mm^2$, and $186\;mm^2$ respectively, Also the mean tumor area in T. canis injected mice in 4 different days was $25.5\;mm^2$ compared to the control group (alum treated) which was $155\;mm^2$. T. gondii parasites and T. canis egg antigens induced inhibition of the tumor growth in the fibrosarcoma mouse model. We need further study to clarify the mechanisms of anti-cancer effects.

Kinetics of IL-23 and IL-12 Secretion in Response to Toxoplasma gondii Antigens from THP-1 Monocytic Cells

  • Quan, Juan-Hua;Zhou, Wei;Cha, Guang-Ho;Choi, In-Wook;Shin, Dae-Whan;Lee, Young-Ha
    • Parasites, Hosts and Diseases
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    • v.51 no.1
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    • pp.85-92
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    • 2013
  • IL-23 and IL-12 are structurally similar and critical for the generation of efficient cellular immune responses. Toxoplasma gondii induces a strong cell-mediated immune response. However, little is known about IL-23 secretion profiles in T. gondii-infected immune cells in connection with IL-12. We compared the patterns of IL-23 and IL-12 production by THP-1 human monocytic cells in response to stimulation with live or heat-killed T. gondii tachyzoites, or with equivalent quantities of either T. gondii excretory/secretory proteins (ESP) or soluble tachyzoite antigen (STAg). IL-23 and IL-12 were significantly increased from 6 hr after stimulation with T. gondii antigens, and their secretions were increased with parasite dose-dependent manner. IL-23 concentrations were significantly higher than those of IL-12 at the same multiplicity of infection. IL-23 secretion induced by live parasites was significantly higher than that by heat-killed parasites, ESP, or STAg, whereas IL-12 secretion by live parasite was similar to those of ESP or STAg. However, the lowest levels of both cytokines were at stimulation with heat-killed parasites. These data indicate that IL-23 secretion patterns by stimulation with various kinds of T. gondii antigens at THP-1 monocytic cells are similar to those of IL-12, even though the levels of IL-23 induction were significantly higher than those of IL-12. The detailed kinetics induced by each T. gondii antigen were different from each other.

Expression of Nucleocapsid Protein Gene of Maaji Virus and Use of the Protein as an Immunodiagnostic Antigen of Hemorrhagic Fever with Renal Syndrome (마지바이러스 Nucleocapsid Protein 유전자의 발현과 신증후 출혈열 진단용 항원으로의 이용)

  • Lee, Pyung-Woo;Kim, Yun-Cheol;Paik, Woo-Hyun
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.77-90
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    • 1996
  • Nucleocapsid protein (NP)which exists in the particle of hantavirus and surrounds the viral RNA genome is one of the major structural proteins and plays role of antigen to elicit the antibody detected predorminantly right after infection of the virus in the patients of hemorragic fever with renal syndrome (HFRS)or experimental animals. NP is important target antigen in serological diagnostic system of HFRS utilizing whole antigens from the native virus particle, such as IFA, ELISA and Western blotting. Therefore, the preparation of this protein in the level of higher quantity and purity is desirasble for developed dianosis of the disease. The purpose of this study is the cloning of NP gene which exists in the S genome segment of Maaji (MAA) virus and expression of the gene to obtain qualified, genetically engineered NP to be utilized as an immunodiagnostic antigen. First of all, for the purpose of amplifing the MAA-NP gene by PCR, the specific primers were built from the known nucleotide sequence of Hantaan viral NP gene. The viral cDNA of the NP gene was synthesized by using the primers and RNase $H^-$ AMV reverse transcriptase. Thereafter, using this cDNA as a template, the NP gene was amplified specifically by Taq DNA polymrerase. The pT7blue (R)T-overhang vector systems were used for cloning of the amplified NP gene. The expression system was consisted of BL21 (DE3)pLysS and pET16b as a host and a plasmid repectively. Into Ndel site of pET16b, NP gene was ligated with cohesive end for the expression. Insertion of NP gene in the plasmid was confirmed by PCR and mini prep methods. For expression, IPTG was used and the expressed protein was characterized by Western blotting. The MAA-NP was expressed as the form of inclusion body (insoluble fraction)and the protein purified by affinity and metal chealating columns reacted specifically with the sera from patients of HFRS as to be tested by ELISA and Western blotting.

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Association Study Analysis of Cluster-of-Differentiation Antigen 9 (CD9) Gene Polymorphism (g.358A>T) for Duroc Boar Post-thawed Semen Motility and Kinematic Characteristics

  • Cho, Eun-Seok;Sa, Soo-Jin;Kim, Ki-Hyun;Lee, Mi-Jin;Ko, Jun-Ho;Kim, Young-Ju;Seol, Kuk-Hwan;Hong, Joon-ki;Kim, Kwang-Sik;Kim, Yong-Min;Woo, Jae-Seok
    • Journal of Embryo Transfer
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    • v.30 no.2
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    • pp.109-114
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    • 2015
  • Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen post-thawed in boar. Recently, polymorphism (g.358A>T) of cluster-of-differentiation antigen 9 (CD9) gene reported to be significant association with MOT. Also, CD9 gene was expressed in the male germ line stem cells is crucial for sperm-egg fusion, and was therefore selected as candidate gene for boar semen. This study was conducted to evaluate the pig SNP (g.358A>T) of CD9 gene as a positional controlling for semen parameters of post-thawed boar semen. To results, the g.358A>T SNP of the CD9 gene was significantly associated with the traits such as MOT, curve linear velocity, straight line velocity, average path velocity and amplitude of lateral head displacement. Particularly, the g.358A>T SNP significantly has the highest association with MOT and animals with AA genotype (p<0.001). Therefore, we suggest that the g.358A>T in the intron 6 region of the porcine CD9 may be used as a molecular marker for Duroc boar Post-thawed semen quality, although its functional effect was not defined yet.

Effects of FUll-FEat Flax Seed, $\alpha$-Tocopherol and Selenium on the Expression of cell Surface Antigen of Broiler Chickens (아마종실과 $\alpha$-Tocopherol, 셀레늄 급여가 육계의 세포표면항원 발현에 미치는 영향)

  • 안종남;채현석;문진산;김동운;권명상;박병성
    • Korean Journal of Poultry Science
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    • v.28 no.3
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    • pp.231-237
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    • 2001
  • To examine the effects of feed additives on the expression of perpheral blood cell surface molecules, phagocytosis and antigen specific antibody formation, broilers were randomly assigned to $T_{1}$ , $T_{2}$ , $T_{3}$ , and $T_{4}$ groups. $T_{1}$ group was fed diet without any additives for 13 weeks, $T_{2}$ was fed diet with full fat flax, $T_{3}$ was fed diet with full fat flax containing $\alpha$-tocopherol, and $T_{4}$ was fed diet with full- fat flax containing $\alpha$-tocopherol and selenium. Since 5 weeks feeding the data were examined by flow cytometry using a panel of monoclonal antibodies. The expression of monocyte in all treated groups was significantly increased, in which the ratio of expression in $T_{3}$ group was especially evident. B cell expression of all treated groups was increased more than 2 fold. The expression of CD4+(helper T cell) cell and CD8+(cytotox$ic^pressor T cell) cell of all treated groups also was increased.ed.

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IL-12 and TNF-α productions from human peripheral blood mononuclear cells in untreated patients with active pulmonary tuberculosis stimulated with 30-kDa or TSP antigen of Mycobacterium tuberculosis H37Rv (결핵균 PPD, 30-kDa 및 TSP 항원에 의한 치료전 폐결핵환자 말초혈액 단핵구의 IL-12 및 TNF-α 생성능)

  • Song, Chang-Hwa;Jo, Eun-Kyeong;Lee, Ji-Suk;Kim, Dae-Su;Lim, Jae-Hyun;Kim, Un-Ok;Nam, Hyeon-Hui;Kim, Hwa-Jung;Paik, Tae-Hyun;Park, Jeong-Kyu
    • IMMUNE NETWORK
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    • v.1 no.3
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    • pp.250-259
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    • 2001
  • To determine if initial infection with Mycobacterium tuberculosis changes the balance of cytokines between T cells and macrophages, we evaluated interferon (IFN)-${\gamma}$), interleukin-12 (IL)-12, and tumor necrosis factor (TNF)-${\alpha}$ productions by peripheral blood mononuclear cells (PBMC) from 15 untreated active pulmonary tuberculosis (TB) patients and 12 healthy tuberculin reactors (HTR). Freshly isolated PBMC were stimulated with Triton X-100 solubilized protein (TSP), 30-kDa or purified protein derivatives (PPD) antigen for 6, 18 and 96 hours. IL-12 p40 production by antigen-stimulated PBMC from TB patients was significantly decreased compared with that in HTR. In addition, IFN-${\gamma}$ production was significantly depressed in TB patients than that in HTR at a 96-hr stimulation. However, TNF-${\alpha}$ production was significantly higher in antigen-stimulated PBMC from TB than that of HTR. A pronounced increase in IFN-${\gamma}$ protein followed neutralization of IL-10 in early TB patients. However, neutralization of TNF-${\alpha}$ did not significantly alter IFN-${\gamma}$ induction in PBMC from TB patients. There were no significantly differences in the cytokine productions among three proteins, TSP, 30-kDa or PPD antigen. These results indicate that development of TB may be strongly associated with dysregulated productions of IL-12, IFN-${\gamma}$ and TNF-${\alpha}$, during the initial immune responses to M. tuberculosis. Further understanding of operative cytokine networks during human immune cell responses to protein antigens of M. tuberculosis may improve strategies for vaccine development.

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Covalent Linkage of IL-12 and Ovalbumin Confines the Effects of IL-12 to Ovalbumin-specific Immune Responses

  • Kim, Tae-Sung;Hwang, Seung-Yong;Yoo, Gyurng-Soo
    • Archives of Pharmacal Research
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    • v.20 no.5
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    • pp.396-403
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    • 1997
  • In order to direct the form of the immune response in an antigen-specific manner, we constructed a fusion protein (OVA/IL12) that contained the T cell-dependent antigen, ovalbumin (OVA), covalently linked to murine interleukin-12 (IL-12). The OVA/IL12 protein was produced in a baculovirus expression system and was purified by anti-OVA immunoaffinity chromatography. The purified OVA/ILI2 protein displayed potent IL-12 bioactivity in an IL-12 proliferation assay. BALB/c mice immunized with the OVA/IL12 protein produced increased quantities of anti-OVA IgG2a antibody compared with mice immunized with recombinant OVA alone. Lymph node cells from the immunized mice with the OVA/IL12 protein produced large amounts of IFN-,Y when restimulated in vitro with OVA, while those from mice immunized with the OVA protein produced little or no IFN-.gamma.. In contrast, immunization with a mixture of OVA and free recombinant IL-12 also induced IFN-.gamma. production, which was not OVA-specific. These studies indicate that the OVA/IL12 fusion protein can induce OVA-specific, Th1-dominated immune responses, and that the covalent linkage of OVA and IL-12 confines the effect of IL-12 to OVA-specific cells.

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Measurement of Antibodies to Varicella-Zoster Virus Using a Virus-Free Fluorescent-Antibody-to-Membrane-Antigen (FAMA) Test

  • Park, Rackhyun;Hwang, Ji Young;Lee, Kang Il;Namkoong, Sim;Choi, Seuk-Keun;Park, Songyong;Park, Hosun;Park, Junsoo
    • Journal of Microbiology and Biotechnology
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    • v.25 no.2
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    • pp.268-273
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    • 2015
  • The fluorescent-antibody-to-membrane-antigen (FAMA) test is regarded as the "gold standard" to detect protective antibodies to varicella-zoster virus (VZV) because of its high sensitivity and specificity. Because the classic FAMA test uses an infectious virus for detection of antibodies to VZV, it is labor-intensive, and also requires special equipment for handling the virus. For this reason, we attempted to develop a simple and safe FAMA assay. Because VZV glycoprotein E (gE) is one of the major VZV glycoproteins, we used the gE protein for the FAMA test (gE FAMA). Here, we demonstrate that overexpression of gE in HEK293T cells can be used to measure antibodies in human serum, and that gE FAMA titers are closely correlated with gpEIA ELISA data. These results indicate that our gE FAMA test has the potential to measure antibodies to VZV.

The Potential Roles of Cyclooxygenase-2 and Matrix Metalloproteinase-9 in Cytomegalovirus-Infected Atherosclerotic Aorta and Coronary Artery

  • Eom, Yong-Bin
    • Biomedical Science Letters
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    • v.9 no.3
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    • pp.123-131
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    • 2003
  • Inflammation appears to have a major role in the development of atherosclerosis. Cyclooxygenase-2 (COX-2) is involved in the inflammatory response via the generation of prostanoids that, in turn, are involved in the production of matrix metalloproteinases (MMPs). This study hypothesized that a vascular infection with cytomegalovirus (CMV) may induce a chronic inflammatory reaction and activated inflammatory cells may express inflammatory mediators such as cyclooxygenase-2 (COX-2) and matrix metalloproteinases-9 (MMP-9). To confirm the hypothesis, the immunohistochemical stains for CMV late antigen, COX-2, MMP-9, macrophage, and T-lymphocyte were performed on CMV-infected atherosclerotic lesions. The immunoreactivity for COX-2 and MMP-9 was evident in all cases of atherosclerosis along with plaques, mainly in macrophages/foamy cells, intimal and medial smooth muscle cells, and endothelial cells of the intima. Within the intima, the increased immunoreactivity for COX-2 and MMP-9 was colocalized to the area stained with CMV late antigen. Sections from control specimens showed no immunoreactivity for CMV late antigen, COX-2 and MMP-9. These data seem to support the hypothesis that CMV may participate in a pathogenetic mechanism for atherogenesis or progression of atherosclerosis.

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Induction of Oral Tolerance by Gamma-Irradiated Ovalbumin Administration

  • Yang, Hui;Lee, Junglim;Seo, Ji Hyun;Oh, Kwang Hoon;Cho, Young Ho;Yoo, Yung Choon
    • Food Science of Animal Resources
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    • v.36 no.1
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    • pp.14-18
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    • 2016
  • Oral administration of soluble antigen can induce peripheral tolerance to the antigen. This study was conducted to evaluate whether gamma-irradiated ovalbumin (OVA) can induce oral tolerance. To investigate this, we administrated intact or irradiated OVA to mice, induced allergic response using intact OVA and alum, then compared humoral and cellular immune responses. Mice treated with gammairradiated OVA had less OVA-specific IgE compared with those who were administered intact OVA. There was no difference in levels of OVA-specific IgG+A+M, IgG1, and IgG2a. Splenocytes of mice administered irradiated OVA showed similar OVA-specific T cell proliferation and secretion of IFN-γ and IL-4. However, there was an increase in IL-2 and a decrease of IL-6 secretion in mice treated with irradiated OVA. These results indicate that gamma-irradiated OVA have similar effects to intact OVA on antigen tolerance.