돼지 卵子의 透明帶에 대한 單一클론抗體生産과 그 特性에 關한 硏究

金鐘培,劉永春,金昌圭,權五中,鄭盛元,鄭吉生 건국대학교 동물자원연구센터 1991 動物資源硏究誌 Vol.16 No.-

本 試驗은 單一클론抗體의 강한 特異性과 抗體性質의 不變性을 이용하여 發生學的 側面에서 哺乳動物 卵子의 透明帶의 機能과 構造를 이해하고, 또한 種特異的인 精子 受容體의 存在 및 生化學的 構造를 규명하기 위한 기초연구로서, 돼지 卵子의 透明帶를 免疫原으로 하여 BABL/c 생쥐로부터 單一클론抗體를 생산하고 그 특성을 구명하였던 바 다음과 같은 결과를 얻었다. 1) 3마리의 BABL/c 생쥐(YⅠ, YⅡ, ZI)에 돼지卵子의 透明帶를 免疫化하고, 複合抗體 生成을 확인한 후 생쥐의 脾臟細胞와 Myeloma(SP2/O-Ag14)를 polyethylene glycol를 融合을 실시한 결과 각각 25.8%, 54.5% 그리고 59.7%의 融合效率을 나타내었으며, ELISQ에 의해 陽性反應을 조사한 결과 각각 17.3%, 32.6% 그리고 6.2% 陽性反應 效率을 나타내었다. 2) YI에서 강한 陽性反應을 보인 6개의 well에 대한 cloning을 실시하고 抗體檢證을 행한 결과 20.8% ∼ 48.4%의 Cloning效率과 54.6% ∼ 82%의 陽性反應 效率을 나타내었다. 3) 강한 陽性反應을 나타낸 항체에 대해 sandwich ELISA法에 의해 isotype을 決定하였던바 2E93C(YⅠ), 3E83E7(YⅠ), 4E3(YⅡ)각각 IgG₂b, IgG₂a, IgM으로 확인되었다. 4) Isotype이 決定된 2E93C9(YⅠ), 3E84E7(YⅠ), 4E3(YⅡ)의 세포를 생쥐의 腹腔에 주사하여 얻은 腹水를 indirect ELISA法에 의해 titer를 決定한 결과 모두 1:400,000 이상의 높은 titer를 나타내었다. 5) 處理區로서 單一클론抗體의 腹水와 對照區로서 normal mouse serum이 각각 2%씩 함유된 배양액속에서 난자를 배양한 후 顯徵鏡下에서 관찰했을 때 對照區에서 배양된 난자의 표면은 정상적인 형태를 나타냈으나 處理區에서 배양된 卵子는 표면에 뚜렷한 沈澱層을 형성하였다. 6) 處理區와 對照區 卵子를 Rabbit anti-mouse IgG-FITC가 1% 함유된 배양액속에서 배양하고 洗滌한 후 螢光顯徵鏡下에서 관찰한 바 處理區의 卵子는 透明帶 주위에서 螢光이 나타났으나, 對照區에서는 나타나지 않았다. This study was carried out ot produce and characterize monoclonal antibodies against porcine zona pellucida, and undertaken as a basic study to develop immunocontraceptive vaccine and to investigate the function of zana pellucida in early fertilization process. The results obtained were summarized as follows: 1. Spleen cells of three BALB/C mice(YⅠ, YⅡ and ZI) which were immunized with porcine zona pelucida were fused with myeloma cells(SP2/O-Ag14) by polyethylene glycol. At the result of fusion, fusion efficiency was 25.8 , 54.5% and 59.7% and positive efficiency 17.3%, 32.6% and 6.2%, respectively. 2. Cloning efficiency was shown to be from 20.8% to 48.4% and positive efficiency of them were 54.6% to 82%. 3. Sub-isotypes of three strong positive antibodies, 2E93C(YⅠ), 3E83E7(YⅠ) and 4E3(YⅡ) were determined by sandwich ELISA method and shown to be as IgG2b, IgH2a and IgM, respectively. 4. The titers of three ascitic fluids containing antibodies, 2E93C9(YⅠ), 3E84E7(YⅠ) and 4E3(YⅡ) were determined by indirect ELISA and all of them showed above 1:400,000. 5. The layer of precipitate was observed on the zona incubated with medium containing 2% ascitic fluid of monoclonal antibody while the eggs treated with 2% normal mouse serum did not. 6. Porcine eggs incubated with medium containing 2% ascitic fluid of monoclonal antibody and followed by subsequent incubation with Rabbit anti-mouse IgG-FITC conjugate showed strong fluorescent light on the zona surface while the zona of normal mouse serum-treated eggs did not show fluorescence.

Caspase-3 activation as a key factor for HBx-transformed cell death

Kim, A.,Kwon, O. S.,Kim, S. O.,He, L.,Bae, E. Y.,Lee, M. S.,Jeong, S. J.,Shim, J. H.,Yoon, D. Y.,Kim, C. H.,Moon, A.,Kim, K. E.,Ahn, J. S.,Kim, B. Y. Blackwell Publishing Ltd 2008 Cell proliferation Vol.41 No.5

<P>Abstract. </P><P><I>Objectives</I>: Nuclear factor-kappa B (NF-&kgr;B) activation has been associated with the tumorigenic growth of hepatitis B virus X protein (HBx)-transformed cells. This study was aimed to find a key target for treatment of HBx-mediated cancers. <I>Materials and methods</I>: NF-&kgr;B activation, endoplasmic reticulum-stress (ER-stress), caspase-3 activation, and cell proliferation were evaluated after Chang/HBx cells permanently expressing HBx viral protein were treated with inhibitors of NF-&kgr;B, proteasome and DNA topoisomerase. <I>Results</I>: Inhibition of NF-&kgr;B transcriptional activity by transient transfection with mutant plasmids encoding Akt1 and glycogen synthase kinase-3&bgr; (GSK-3&bgr;), or by treatment with chemical inhibitors, wortmannin and LY294002, showed little effect on the survival of Chang/HBx cells. Furthermore, I&kgr;Bα (S32/36A) mutant plasmid or other NF-&kgr;B inhibitors, 1-pyrrolidinecarbonidithioic acid and sulphasalazine, were also shown to have little effect on the cell proliferation. By contrast, proteasome inhibitor-1 (Pro1) and MG132 enhanced the HBx-induced ER-stress response and the subsequent activation of caspase-12, -9 and -3 and reduced cell proliferation. Camptothecin (CPT), however, triggered activation of caspase-3 without induction of caspase-12, and reduced cell proliferation. In addition, CPT-induced cell death was reversed by pre-treatment with z-DEVD, a caspase-3-specific inhibitor. <I>Conclusions</I>: Detailed exploitation of the regulators of caspase-3 activation could open the gate for finding an efficient target for development of anticancer therapeutics against HBx-transformed hepatocellular carcinoma.</P>

The inability of Bacillus licheniformis perR mutant to grow is mainly due to the lack of PerR-mediated fur repression

Kim, J. H.,Yang, Y. M.,Ji, C. J.,Ryu, S. H.,Won, Y. B.,Ju, S. Y.,Kwon, Y.,Lee, Y. E.,Youn, H.,Lee, J. W. MICROBIOLOGICAL SOCIETY OF KOREA 2017 JOURNAL OF MICROBIOLOGY -SEOUL- Vol. No.

<P>PerR, a member of Fur family protein, is a metal-dependent H2O2 sensing transcription factor that regulates genes involved in peroxide stress response. Industrially important bacterium Bacillus licheniformis contains three PerR-like proteins (PerR(BL), PerR2, and PerR3) compared to its close relative Bacillus subtilis. Interestingly, unlike other bacteria including B. subtilis, no authentic perR (BL) null mutant could be established for B. licheniformis. Thus, we constructed a conditional perR (BL) mutant using a xylose-inducible promoter, and investigated the genes under the control of PerR(BL). PerR(BL) regulon genes include katA, mrgA, ahpC, pfeT, hemA, fur, and perR as observed for PerR(BS). However, there is some variation in the expression levels of fur and hemA genes between B. subtilis and B. licheniformis in the derepressed state. Furthermore, katA, mrgA, and ahpC are strongly induced, whereas the others are only weakly or not induced by H2O2 treatment. In contrast to the B. subtilis perR null mutant which frequently gives rise to large colony phenotype mainly due to the loss of katA, the suppressors of B. licheniformis perR mutant, which can form colonies on LB agar, were all catalase-positive. Instead, many of the suppressors showed increased levels of siderophore production, suggesting that the suppressor mutation is linked to the fur gene. Consistent with this, perR fur double mutant could grow on LB agar without Fe supplementation, whereas perR katA double mutant could only grow on LB agar with Fe supplementation. Taken together, our data suggest that in B. licheniformis, despite the similarity in PerR(BL) and PerR(BS) regulon genes, perR is an essential gene required for growth and that the inability of perR null mutant to grow is mainly due to elevated expression of Fur.</P>

Early Regulation of Viral Infection Reduces Inflammation and Rescues Mx-positive Mice from Lethal avian Influenza Infection

Song, M.S.,Cho, Y.H.,Park, S.J.,Pascua, P.N.Q.,Baek, Y.H.,Kwon, H.I.,Lee, O.J.,Kong, B.W.,Kim, H.,Shin, E.C.,Kim, C.J.,Choi, Y.K. American Association of Pathologists and Bacteriol 2013 The American journal of pathology Vol.182 No.4

Differing sensitivity of influenza A viruses to antiviral effects of the Myxovirus resistance (Mx) protein implies varying global gene expression profiles in the host. The role of Mx protein during lethal avian influenza (AI) virus infection was examined using Mx1-deficient C57BL/6 (B6-Mx1<SUP>-/-</SUP>) and congenic Mx1-expressing (B6-Mx1<SUP>+/+</SUP>) mice infected with a virulent, mouse-adapted avian H5N2 Ab/Korea/ma81/07 (Av/ma81) virus. After infection, B6-Mx1<SUP>+/+</SUP> mice were completely protected from lethal AI-induced mortality, and exhibited attenuated clinical disease and reduced viral titers and pathology in the lungs, compared with B6-Mx1<SUP>-/-</SUP> mice. Transcriptional profiling of lung tissues revealed that most of the genes up-regulated after infection are involved in activation of the immune response and host defense. Notably, more abundant and sustained expression of cytokine/chemokine genes was observed up to 3 dpi in B6-Mx1<SUP>-/-</SUP> mice, and this was associated with excessive induction of cytokines and chemokines. Consequently, massive infiltration of macrophages/monocytes and granulocytes into lung resulted in severe viral pneumonia and potentially contributed to decreased survival of B6-Mx1<SUP>-/-</SUP> mice. Taken together, our data show that dysregulated gene transcriptional activity corresponded to persistent induction of cytokine/chemokines and recruitment of cytokine-producing cells that promote inflammation in B6-Mx1<SUP>-/-</SUP> mouse lungs. Thus, we provide additional evidence of the interplay of genetic, molecular, and cellular correlates governed by the Mx1 protein that critically determine disease outcome during lethal AI virus infection.

Hardening of Bi-Te based alloys by dispersing B₄C nanoparticles

Jung, S.J.,Park, S.Y.,Kim, B.K.,Kwon, B.,Kim, S.K.,Park, H.H.,Kim, D.I.,Kim, J.Y.,Hyun, D.B.,Kim, J.S.,Baek, S.H. Elsevier Science 2015 Acta materialia Vol.97 No.-

Thermoelectric devices have attracted a great attention for renewable energy harvesters and solid-state coolers. For practical applications, the mechanical properties of thermoelectric materials become critical for the device reliability, a persistent performance with a long time and high operation cycles. Bi-Te based single-crystals, mostly used in commercial thermoelectric devices, are intrinsically brittle with weak van der Waals bonding, often leading to device failures such as crack and debonding during fabrication and operation. Thus, it is highly desirable to enhance the mechanical property of Bi-Te based alloys as well as the thermoelectric property. Here, we investigate the effect of B<SUB>4</SUB>C nanoparticles (less than 0.5wt%) dispersed in p-type Bi<SUB>0.4</SUB>Sb<SUB>1.6</SUB>Te<SUB>3</SUB> matrix on the mechanical properties. X-ray diffraction (XRD) result confirms that B<SUB>4</SUB>C-dispersed Bi<SUB>0.4</SUB>Sb<SUB>1.6</SUB>Te<SUB>3</SUB> has a single phase. We observe that the grain size of Bi<SUB>0.4</SUB>Sb<SUB>1.6</SUB>Te<SUB>3</SUB> becomes decreased with the B<SUB>4</SUB>C nanoparticle concentration by electron backscatter diffraction (EBSD) technique. Hardness, Young's modulus, and flexural strength of B<SUB>4</SUB>C-dispersed Bi<SUB>0.4</SUB>Sb<SUB>1.6</SUB>Te<SUB>3</SUB> are enhanced, compared to the B<SUB>4</SUB>C-free Bi<SUB>0.4</SUB>Sb<SUB>1.6</SUB>Te<SUB>3</SUB> polycrystals. On the other hand, the thermoelectric figure-of-merit of B<SUB>4</SUB>C-dispersed Bi<SUB>0.4</SUB>Sb<SUB>1.6</SUB>Te<SUB>3</SUB> is almost identical to that of the pure Bi<SUB>0.4</SUB>Sb<SUB>1.6</SUB>Te<SUB>3</SUB>. Such enhancements of the mechanical properties of the B<SUB>4</SUB>C-dispersed Bi<SUB>0.4</SUB>Sb<SUB>1.6</SUB>Te<SUB>3</SUB> are attributed to the grain boundary hardening and second-phase hardening. Beyond thermoelectric materials, our result implies that the grain refinement by nanoparticle dispersion is a simple and promising way to strengthen the mechanical properties of other brittle materials with layered structure.

Efficacy of HVT-IBD vector vaccine compared to attenuated live vaccine using in-ovo vaccination against a Korean very virulent IBDV in commercial broiler chickens

Roh, J.-H.,Kang, M.,Wei, B.,Yoon, R.-H.,Seo, H.-S.,Bahng, J.-Y.,Kwon, J.-T.,Cha, S.-Y.,Jang, H.-K. Elsevier 2016 Poultry science Vol.95 No.5

<P>The production performance, efficacy, and safety of two types of vaccines for infectious bursal disease virus (IBDV) were compared with in-ovo vaccination of Cobb 500 broiler chickens for gross and microscopic examination of the bursa of Fabricius, bursa/body weight (b/B) ratio, flow cytometry, and serologic response to Newcastle disease virus (NDV) vaccination. One vaccine was a recombinant HVT-IBD vector vaccine (HVT as for herpesvirus of turkeys) and the other was an intermediate plus live IBDV vaccine. A significant difference was detected at 21 d. Eight of 10 chickens that received the IBDV live vaccine had severe bursal lesions and a relatively low b/B ratio of 0.95, and an inhibited NDV vaccine response. On the other hand, the HVT-IBD vector vaccine resulted in mild bursal lesions and a b/B ratio of 1.89. Therefore, the live vaccine had lower safety than that of the HVT-IBD vector vaccine. To determine the protective efficacy, chickens were intraocularly challenged at 24 d. Eight of 10 chickens in the IBDV live vaccination group showed gross and histological lesions characterized by hemorrhage, cyst formation, lymphocytic depletion, and a decreased b/B ratio. In contrast, the HVT-IBD vector vaccinated chickens showed mild gross and histological lesions in three of 10 chickens with a b/B ratio of 1.36, which was similar to that of the unchallenged controls. Vaccinated chickens showed a significant increase in IBDV antibody titers, regardless of the type of vaccine used. In addition, significantly better broiler flock performance was observed with the HVT-IBD vector vaccine compared to that of the live vaccine. Our results revealed that the HVT-IBD vector vaccine could be used as an alternative vaccine to increase efficacy, and to have an improved safety profile compared with the IBDV live vaccine using in-ovo vaccination against the Korean very virulent IBDV in commercial broiler chickens.</P>

Study on the HDDR Characteristics of the Nd-Fe(-Co)-B(-Ga-Zr)-type Alloys

S. W. Shon,H. W. Kwon,D. I. Kang,Yoon. B. Kim,W. Y. Jeung 한국자기학회 1999 Journal of Magnetics Vol.4 No.4

The HDDR characteristics of the Nd-Fe-B-type isotropic and anisotropic HDDR alloys were investigated using three types of alloys: alloy A (Nd_(12.6)Fe_(81.4)B_6), alloy B (Nd_(l2.6)Fe_(81.3)B_6Zr_(0.l)), and alloy C (Nd_(12.6)Fe_(68.8)CO_(11.5)B_6Ga_(1.0)Zr_(0.1)). The alloy A is featured with the isotropic HDDR character, while alloy Band C are featured with the anisotropic HDDR character. Hydrogenation and disproportionation characteristics of the alloys were examined using DTA under hydrogen gas. Recombination characteristics of the alloys were examined by observing the coercivity variation as a function of recombination time. The present study revealed that the alloy C exhibits slightly higher hydrogenation and disproportionation temperatures compared to the alloy A and B. Recombination of the anisotropic alloy Band C takes place more rapidly with respect to the isotropic alloy A. The intrinsic coercivities of the recombined materials rapidly increased with increasing the recombination time and then showed a peak, after which the coercivities decreased gradually. The degraded coercivity was, however, recovered significantly on prolonged recombination treatment. Compared with the isotropic HDDR alloy A the anisotropic HDDR alloy B and C are notable for their greater recovery of coercivity. The significant recovery of coercivity was accounted for in terms of the development of well-defined smooth grain boundary between the recombined grains on prolonged recombination.

Aberrant ventral striatal responses during incentive processing in unmedicated patients with obsessive–compulsive disorder

Jung, W. H.,Kang, D.‐,H.,Han, J. Y.,Jang, J. H.,Gu, B.‐,M.,Choi, J.‐,S.,Jung, M. H.,Choi, C.‐,H.,Kwon, J. S. Blackwell Publishing Ltd 2011 Acta psychiatrica Scandinavica Vol.123 No.5

<P>Jung WH, Kang D‐H, Han JY, Jang JH, Gu B‐M, Choi J‐S, Jung MH, Choi C‐H, Kwon JS. Aberrant ventral striatal responses during incentive processing in unmedicated patients with obsessive–compulsive disorder.</P><P><B>Objective: </B> Obsessive–compulsive disorder (OCD) is characterized by the dysfunction of control and reward mechanisms. However, only few neuroimaging studies of OCD have examined the reward processing. We examined the neural responses during incentive processing in OCD.</P><P><B>Method: </B> Twenty unmedicated patients with OCD and 20 age‐, sex‐, and IQ‐matched healthy controls underwent functional magnetic resonance imaging while performing a modified monetary incentive delay task.</P><P><B>Results: </B> Compared with controls, patients with OCD showed increased ventral striatal activation in the no‐loss minus loss outcome contrast and a significant positive correlation between the ventral striatal activation and compulsion symptom severity. In addition, patients with OCD showed increased activations in the frontostriatal regions in the gain minus no‐gain outcomes contrast. During loss anticipation, patients with OCD showed less activations in the lateral prefrontal and inferior parietal cortices. However, during gain anticipation, patients with OCD and healthy controls did not differ in the ventral striatal activation.</P><P><B>Conclusion: </B> These findings provide neural evidence for altered incentive processing in unmedicated patients with OCD, suggesting an elevated sensitivity to negatively affect stimuli as well as dysfunction of the ventral striatum.</P>

Structural aspect of B*4811 molecule, a novel HLA-B*48 allele identified by sequence-based typing

Kwon, O.-J.,Hwang, S.-H.,Heo, Y.-S.,Hur, S.-S.,Oh, H.-B. Munksgaard 2005 Tissue antigens Vol.66 No.6

<i>HLA‐B*40:02:18</i>, a new <i>HLA‐B*40</i> allele

Ko, S.‐,Y.,Oh, H.‐,B.,Jun, J.‐,H.,Kwon, O.‐,J. Munksgaard 2014 Tissue antigens Vol.83 No.6

<P>The new allele <I>B*40:02</I>:<I>18</I> showed one nucleotide difference with <I>B*40:02:01</I> at codon 111 (CGC/CGT).</P>