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Rhie, Gi-Eun,Jung, Hae-Mi,Park, Jungchan,Kim, Bong-Su,Mekalanos, John J. Oxford University Press 2008 FEMS immunology and medical microbiology Vol.52 No.1
<P>The most widely used oral whole-cell-recombinant B subunit cholera vaccine contains the nontoxic cholera toxin B subunit (CTXB) and either heat- or formalin-killed Vibrio cholerae O1 strains. Vibrio cholerae O1 strains in the vaccine provide antibacterial immunity, and CTXB contributes to the vaccine's efficacy by stimulating production of anti-CTXB antibody. Various attempts have been made to increase CTXB production. In this study, the mariner-FRT transposon delivery system developed by Chiang and Mekalanos was used to place the ctxB gene under the control of a strong chromosomal promoter in a nontoxigenic V. cholerae El Tor strain, M7922. The expression level of CTXB in transposon insertion mutant clones was screened by ganglioside-dependent enzyme-linked immunosorbent assay. Among CTXB-producing V. cholerae clones that were isolated, M7922-C1 produced the highest amount of CTXB (3.17+/-1.69 microg mL(-1)). M7922-C1 harbors a single insertion of ctxB into VC0972, which encodes a putative porin protein. Although the level of CTXB expression in this strain was not exceptionally high, this study indicates the possibility of using this delivery system to construct vaccine strains that overexpress specific antigens.</P>
Rhie, Gi Eun,Chung, Gyung Tae,Lee, Yong Jin,Sung, Won Keung,Oh, Hee Bok 한국미생물 · 생명공학회 2000 Journal of microbiology and biotechnology Vol.10 No.1
We have earlier reported on the cloning and identification of bft-k from an enterotoxigenic strain of Bacteroides fragilis 419, which was isolated from the blood of a Korean patient who suffered from systemic infections [4, 5]. The bft-k gene encodes a 397-amino-acids metalloprotease enterotoxin, and the protein has been identified as a new isoform of B. fragilis enterotoxins (BFTs), which are cytopathic to intestinal epithelial cells to induce fluid secretion and tissue damage in ligated intestinal loops [4, 6, 18, 20]. This report describes the cloning and sequencing of the enterotoxin pathogenicity islet of B. fragilis 419 which contains the bft-k gene. The cloned enterotoxin pathogenicity islet was found to have 6,045bp in length and to contain 12-bp direct repeats near its end. In the pathogenicity islet, in addition to the BFT-K, two putative open reading frames (ORFs) were identified; (1) the t-3 gene encoding a 396-aminoacids protein of a putative metalloprotease; (2) the third gene encoding an ORF of a 59-amino-acids protein, whose function has not yet been characterized. The expression of the t-3 gene in B. fragilis 419 was verified by western blot analysis.
Ahn, Bo-Eun,Bae, Hee-Won,Lee, Hae-Ri,Woo, Sun-Je,Park, Ok-Kyu,Jeon, Jun Ho,Park, Jungchan,Rhie, Gi-eun Elsevier 2019 Biochemical and biophysical research communication Vol.509 No.2
<P><B>Abstract</B></P> <P>Since <I>Bacillus anthracis</I> is a high-risk pathogen and a potential tool for bioterrorism, numerous therapeutic methods including passive immunization have been actively developed. Using a human monoclonal antibody phage display library, we screened new therapeutic antibodies for anthrax infection against protective antigen (PA) of <I>B. anthracis</I>. Among 5 selected clones of antibodies based on enzyme-linked immunosorbent assay (ELISA) results, 7B1 showed neutralizing activity to anthrax lethal toxin (LT) by inhibiting binding of the domain 4 of PA (PD4) to its cellular receptors. Through light chain shuffling process, we improved the productivity of 7B1 up to 25 folds. The light chain shuffled 7B1 antibody showed protective activity against LT both <I>in vitro</I> and <I>in vivo</I>. Furthermore, the antibody also conferred protection of mice from 3 × LD<SUB>50</SUB> challenges of fully virulent anthrax spores. Our result expands the possibility of developing a new therapeutic antibody for anthrax cure.</P> <P><B>Highlights</B></P> <P> <UL> <LI> We screened a novel therapeutic antibody for anthrax using phage display library. </LI> <LI> The selected antibody recognizes domain 4 of protective antigen of anthrax toxin. </LI> <LI> The selected antibody showed anthrax toxin-neutralizing efficacy in mouse model. </LI> </UL> </P>
Shin, Mi Hee,Rhie, Gi-eun,Kim, Yeon Kyung,Park, Chi-Hyun,Cho, Kwang Hyun,Kim, Kyu Han,Eun, Hee Chul,Chung, Jin Ho Blackwell Publishing 2005 The Journal of investigative dermatology Vol.125 No.2
<P>To understand the molecular alterations occurring during the aging process, we compared mitogen-activated protein (MAP) kinase activities in the intrinsically aged and photoaged skins in the same individuals. Furthermore, we investigated the molecular events related to MAP kinase changes in intrinsically aged and photoaged skins. We found that extracellular-signal-regulated kinase (ERK) activity in photoaged skin was reduced, and that the activities of c-Jun N-terminal kinase (JNK) and p38 kinase were increased compared with intrinsically aged skin in the same individuals. Phospho-c-Jun levels and activator protein 1 activities in photoaged skin were also higher than in intrinsically aged skin. Moreover, catalase activity was found to be much reduced in primary dermal fibroblasts from photoaged skin, and as a result, H<SUB>2</SUB>O<SUB>2</SUB> accumulated more in primary dermal fibroblasts in photoaged skin. In addition, treating primary dermal fibroblasts from photoaged skin with catalase reduced H<SUB>2</SUB>O<SUB>2</SUB> levels, reversed aging-dependent MAP kinase changes, and inhibited matrix metalloproteinase (MMP)-1 expression. Our results indicate that the accumulation of reactive oxygen species due to catalase attenuation may be a critical aspect of the MAP kinase signaling changes that may lead to skin aging and photoaging in human skin <I>in vivo</I>. Thus, the induction and regulation of endogenous antioxidant enzymes including catalase may offer a strategy for preventing and treating skin aging.</P>
LEE, GUN YOUNG,RHIE, GI-EUN,CHUNG, GYUNG TAE,SUNG, WON-KEUN,OH, HEE-BOK 한국미생물 · 생명공학회 2003 Journal of microbiology and biotechnology Vol.13 No.2
We earlier reported the identification of bft-k, t-3, and a third ORF from an enterotoxigenic strain of Bacteroides fragilis 419, which was isolated from the blood of a Korean patient suffering from systemic infections. In the present study, the deleted fragments of the t-3 and the bft-k genes from B. fragilis 419 were cloned into suicide vector pJST55 and used to create a mutant with chromosomal disruption of the t-3 and bft-k genes. Structures of the selected mutants, DMP-2 and DBT-4, were found to be intermediate forms that integrated the suicide vector into the chromosome. t-3 disrupted DNP-2 and bft-k disrupted DBT-4 did not react with polyclonal antibodies against T-3 or BFT-K, and had no biological activity in HT29/C, cells.
( Hae-ri Lee ),( Jun Ho Jeon ),( Gi-eun Rhie ) 한국미생물 · 생명공학회 2017 Journal of microbiology and biotechnology Vol.27 No.5
The poly-γ-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.
위서리,한지은,Yeon Hee Kim,Gi-eun Rhie,이나경 대한약학회 2013 Archives of Pharmacal Research Vol.36 No.4
Anthrax is an acute infectious disease causedby Bacillus anthracis. We previously reported that theadjuvant CIA06B, which consists of TLR4 agonist CIA05and aluminum hydroxide (alum), enhanced the immuneresponse to anthrax protective antigen (PA) in mice. Thisstudy was carried out to determine whether CIA06B canenhance long-term immune responses to PA in mice. BALB/c mice were immunized intramuscularly three timesat 2-week intervals with recombinant PA alone or PAcombined with alum or CIA06B. At 8 and 24 weeks postimmunization,the immunological responses includingserum anti-PA IgG antibody titer, toxin-neutralizing antibodytiter, splenic cytokine secretion and the frequency ofPA-specific memory B cells were assessed. Compared withmice injected with PA alone or PA plus alum, miceinjected with PA plus CIA06B had higher titers of serumanti-PA IgG antibodies, and higher frequencies of PAspecificmemory B cells and interferon-c secreting cells. Furthermore, anti-PA antibodies induced by CIA06B weremore effective in neutralizing anthrax toxin. These resultsdemonstrated that CIA06B is capable of providing longtermimmunity when used as an adjuvant in a PA-basedanthrax vaccine.