• Title/Summary/Keyword: Antibody Response

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Characterization of Monoclonal Antibodies against Heavy and Light Chains of Flounder (Paralichthys olivaceus) Immunoglobulin

  • Jang, Han-Na;Woo, Jong-Kyu;Cho, Young-Hye;Kyong, Seo-Bong;Choi, Sang-Hoon
    • BMB Reports
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    • v.37 no.3
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    • pp.314-319
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    • 2004
  • Flounder (Paralichthys olivaceus) Immunoglobulins (Igs) were purified from the serum of mouse IgG-immunized flounder by using affinity chromatography. Under denaturing conditions in SDS-PAGE, the flounder Igs appeared to be composed of 2 heavy (H) chains (72 and 77 kDa) and two light (L) chains (26 and 28 kDa). Monoclonal antibodies (MAbs) were produced by the fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells that were previously sensitized against affinity-purified flounder Igs. In a Western blot analysis, the produced MAbs, FIM511, FIM519, and FIM562 recognized both the 72 and 77 kDa H chains, 26 kDa, and 28 kDa L chain, respectively. Mouse antiserum against flounder Igs reacted more strongly with the L chain of 28 kDa than with 26 kDa, suggesting that the 28 kDa molecule is more immunogenic than the 26 kDa L chain molecule. In a FACS analysis, the ratios of the Ig+ cell population in the flounder head kidney and spleen cells were 49% and 24%, respectively. Unexpectedly, however, the ratios of the Ig+ B-like cell population in the flounder were not significantly augmented, even after the immunization of an immunogenic antigen. This suggests that the humoral immune response in fish could be considerably different from that in mammals. The produced MAbs in this study would be useful in characterizing flounder Ig+ B-like cells and in developing flounder Ig detecting an immunoassay system.

STUDIES ON IMMUNOTOXIC POTENTIAL OF METHAMPHETAMINE (MA) IN Balb/C MICE II. The Functional Alteration of Effector Cells in Immune System

  • Lim, Chae-Woong;Rim, Byung-Moo;Lee, Ho-Il;Kim, Sang-Ho
    • Toxicological Research
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    • v.11 no.1
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    • pp.15-21
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    • 1995
  • Several populations of lymphocytes possess receptors for autonomic neurotransmitter, which make lymphocytes susceptible to autonomic stimulation. This study was to evaluate the functional alternation of effector cells of the immune system. Female Balb/C mice, 15-20 g, were injected with MA subcutaneously under various conditions. Mixed lymphocyte reaction (MLR) showed certain T cell subsets were affected by MA. The level of interleukin-2 (IL-2) production was inhibited due to a defect in expression of the IL-2 receptor. In mice injected with 20 mg MA/kg, 1 day before assay, phagocytosis of peritoneal macrophages showed $14.07\pm3%$, which was similar degree to 5 mg MA/kg treatment for 4 consecutive days. Phagocytosis was almost recovered to that of control after 4 day in 20 mg/kg injected mice. Maximum inhibition of plaque forming cell (PFC) occurred when MA was given early, indicating the inductive time point of antibody production was affected. The cortisol level increased in the MA treated group (0.05, 0.20, and $0.08{\mu}g$/dl for control, low, and high dose-MA treated mice, respectively). Based on these results, MA has general suppression effects on the immune systems by functional alteration of effector cells. Considering the increment of serum cortisol levels, MA partially impacts the neuroendocrine system to lead to failure of immune response.

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Epidemiological Investigation of an Outbreak of Hepatitis A at a Residential Facility for the Disabled, 2011

  • Lim, Hyun-Sul;Choi, Kumbal;Lee, Saerom
    • Journal of Preventive Medicine and Public Health
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    • v.46 no.2
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    • pp.62-73
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    • 2013
  • Objectives: An outbreak of hepatitis A occurred at a residential facility for the disabled in July 10, 2011. This investigation was carried out to develop a response plan, and to find the infection source of the disease. Methods: A field epidemiologist investigated the symptoms, vaccination histories, living environments, and probable infection sources with 51 residents and 31 teachers and staff members. In July 25, 81 subjects were tested for the hepatitis A virus antibody, and specimens of the initial 3 cases and the last case were genetically tested. Results: Three cases occurred July 10 to 14, twelve cases August 3 to 9, and the last case on August 29. Among the teachers and staff, no one was IgM positive (on July 25). The base sequences of the initial 3 and of the last case were identical. The vehicle of the outbreak was believed to be a single person. The initial 3 patients were exposed at the same time and they might have disseminated the infection among the patients who developed symptoms in early August, and the last patient might have, in turn, been infected by the early August cases. Conclusions: The initial source of infection is not clear, but volunteers could freely come into contact with residents, and an infected volunteer might have been the common infection source of the initial patients. Volunteers' washing their hands only after their activity might be the cause of this outbreak. Although there may be other possible causes, it would be reasonable to ask volunteers to wash their hands both before and after their activities.

Modulation of Bee Venom on Th1/Th2 Cell Lineage Development (봉독 추출액이 helper T cell 분화에 미치는 영향)

  • Ko Eun Jung;Nam Sang Soo;Hong Moo Chang;Shin Min Kyu;Bae Hyun Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.5
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    • pp.1347-1355
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    • 2004
  • In this study, the immunological effect of a traditional Korea herbal acupuncture, that has been widely used for the treatment of various immunological disorders including inflammation in Korea, was examined in vitro and in vivo. In our previous study demonstrated that BV increased the expression of IFN-γ mRNA, that plays pivotal role in T cell response. This study was designated to evaluate the effect of BV on helper T cell development by monitoring Th1/Th2 specific cytokine secretion patterns in artificially induced Th1/Th2 polarized condition and in vivo. The results demonstrated that BV didn't have mitogenic effects on the unstimulated CD4+ T cell, but increased the CD4+ T cell proliferation upon activation with anti-CD3/CD28 antibody. The Th1 cells were over-populated dramatically in Th1 driven condition with BV treatment, while the Th2 cells were increased slightly in Th2 skewed condition. Furthermore, under Th1-skewed conditions, the level of IFN-γ was considerably increased with BV treatment. Besides, the expression of T-bet, a transcription factor that plays pivotal role in Th1 lineage programming, was increased with BV treatment. The expressions of IFN-γ and T-bet were also significantly increased in vivo. The results that Th1 specific cytokine secretion were considerably increased and Th2 specific cytokine secretion were not significantly changed in vitro and in vivo indicated that BV enhances Th1 lineage development, Therefore, these results suggest that BV might be desirable agent for correction of Th1 dominant pathological disorders.

Evaluation of the recent live vaccination effects against Newcastle disease under field conditions (최근 야외농장에서 실시하고 있는 뉴캣슬병 생독백신 접종효능에 대한 평가)

  • Song, Chang-seon;Lee, Youn-jeong;Han, Myung-guk;Seong, Hwan-woo;Kang, Kyung-soo;Lee, Joong-bok;Kim, Jae-hak
    • Korean Journal of Veterinary Research
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    • v.40 no.3
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    • pp.563-573
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    • 2000
  • Periodic outbreaks of Newcastle disease (ND) caused by velogenic viscerotropic ND virus (vvNDV) has become a major concern in Korea nowadays. Throughout last epidemic, the winter season in 2000, most chicken flocks infected early, under 2-4 weeks of age, showed high mortality up to 50-100%. Serum samples collected from 201 breeder, 284 layer and 112 broiler chicken flocks were examined to evaluate the efficacy of various vaccination methods and programs routinely used for mass vaccination in the field poultry farms. Despite repeated live vaccination, most poultry flocks vaccinated by drinking water route using nipple water supply system failed to produce solid active immune response to NDV during the growing time. In the present study, we applied the spray vaccination technique using Ulvavac or Desvac sprayer to the experimental poultry flocks and examined the efficacy of live vaccination effects induced by it under field condition. Measurable antibody to NDV as well as early protection against vvNDV challenge were found in poultry flocks vaccinated by spray route. Further, we did not found significant post vaccination reactions caused by spray vaccination if properly administered. These data indicate that the spray vaccination will be safe and reliable mass vaccination method for the prevention of ND.

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Effects of Various Field Coccidiosis Control Programs on Host Innate and Adaptive Immunity in Commercial Broiler Chickens

  • Lee, Kyung-Woo;Lillehoj, Hyun S.;Jang, Seung-I.;Lee, Sung-Hyen
    • Korean Journal of Poultry Science
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    • v.39 no.1
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    • pp.17-25
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    • 2012
  • Coccidiosis control programs such as vaccines or in-feed anticoccidials are commonly practiced in the poultry industry to improve growth performance and health of commercial broiler chickens. In this study, we assessed the effects of various coccidiosis control programs (e.g., in ovo vaccination, synthetic chemicals, and antibiotic ionophores) on immune status of broiler chickens vaccinated against infectious bronchitis virus and Newcastle disease virus (ND) and raised on an Eimeria-contaminated used litter. In general, the levels of ${\alpha}$-1-acid glycoprotein, an acute phase protein, were altered by the treatments when measured at 34 days of age. Splenocyte subpopulations and serum antibody titers against ND were altered by various coccidiosis control programs. In-ovo-vaccinated chickens exhibited highest mitogenic response when their spleen cells were stimulated with concanavalin A (Con A) at 7 days of age. It is clear from this study that the type of coccidiosis control program influenced various aspects of innate and adaptive immune parameters of broiler chickens. Further studies will be necessary to delineate the underlying relationship between the type of coccidiosis control program and host immune system and to understand the role of other external environmental factors such as gut microbiota on host-pathogen interaction in various disease control programs.

Purification and Characterization of Mitogen -Activated Protein (MAP) Kinase from Mammalian Tissue Cells (동물 조직세포로부터 Mitogen-activated Protein (MAP) Kinase의 분리 및 성격규명)

  • 김태우;정동주;김윤석
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.21-30
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    • 1996
  • MAP kinases are a family of serine/threonine specific protein kinases becoming activated in response to different proliferative stimuli by phosphorylation at both threonine and tyrosine residue. Present study shows that MAP kinase was purified from P388 murine leukema cells by SP sephadex C-50, phenyl superose and Mono Q column chromatography and identified with anti-ERKl antibody by western blotting. Immnublotting analysis to the crude extract of P388 cell lysate shows 44 kD and other minor bands but partial purified fraction eluted from phenyl supherose column have 44kD and 66 kD isoform. Subcloned GST-fusion protein from N-terminal of $p56^{kk}$ was tested as a substrate for MAP kinase phosphorylation. It was showed that the wild type and mutant forms(S42A) were fully phosporylated by purified MAP kinase fraction as com-pare with the other mutant form(S59A). This finding suggest that those GST-fusion proteins may be used as substrate for the in vitro test of MAP kinase.

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Protective Effects of Diallyl Sulfide against Thioacetamide-Induced Toxicity: A Possible Role of Cytochrome P450 2E1

  • Kim, Nam Hee;Lee, Sangkyu;Kang, Mi Jeong;Jeong, Hye Gwang;Kang, Wonku;Jeong, Tae Cheon
    • Biomolecules & Therapeutics
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    • v.22 no.2
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    • pp.149-154
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    • 2014
  • Effects of diallyl sulfide (DAS) on thioacetamide-induced hepatotoxicity and immunotoxicity were investigated. When male Sprague-Dawley rats were treated orally with 100, 200 and 400 mg/kg of DAS in corn oil for three consecutive days, the activity of cytochrome P450 (CYP) 2E1-selective p-nitrophenol hydroxylase was dose-dependently suppressed. In addition, the activities of CYP 2B-selective benzyloxyresorufin O-debenzylase and pentoxyresorufin O-depentylase were significantly induced by the treatment with DAS. Western immunoblotting analyses also indicated the suppression of CYP 2E1 protein and/or the induction of CYP 2B protein by DAS. To investigate a possible role of metabolic activation by CYP enzymes in thioacetamide-induced hepatotoxicity, rats were pre-treated with 400 mg/kg of DAS for 3 days, followed by a single intraperitoneal treatment with 100 and 200 mg/kg of thioacetamide in saline for 24 hr. The activities of serum alanine aminotransferase and aspartate aminotransferase significantly elevated by thioacetamide were protected in DAS-pretreated animals. Likewise, the suppressed antibody response to sheep erythrocytes by thioacetamide was protected by DAS pretreatment in female BALB/c mice. Taken together, our present results indicated that thioacetamide might be activated to its toxic metabolite(s) by CYP 2E1, not by CYP 2B, in rats and mice.

The Comparative Study of IgG, IgM, and IgA in Laboratory Animal Administrated Red-ginseng, Using Immunoglobulin Productivity Assay (Immunoglobulin productivity assay를 이용(利用)한 홍삼투여(紅蔘投與) 실험동물(實驗動物)의 IgG, IgM, IgA 비교(比較) 연구(硏究))

  • Lee, Beom-Jun;So, Hyung-Jin;Kim, Jae-Wan;Lew, Jae-Hwan
    • The Journal of Internal Korean Medicine
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    • v.28 no.4
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    • pp.886-895
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    • 2007
  • Objective : The immune system is a complex of systems, all of which work together to clear infection from the body. In Korea, red ginsenghas been one of the herbs most widely used to enhance the immune system for thousand of years. More recently, red ginseng has been reported to have many positive effects on the immune system. The purpose of this study was evaluate the effects of Korean red ginseng and Chinese red ginseng on IgG, IgM, and IgA, using immunoglobulin productivity assay. Methods : Male SD rats were separated into 3 groups. We administered Korean red ginseng (KRG) to one group and Chinese red ginseng (CRG) to another, with normal saline for the Control group consecutively and orally for 3 months. The dose of red ginseng was 500mg per day, as a powder with soluble water. Immunoglobulin levels from spleen cell were estimated by ELISA kit. Results : In immunoglobulin productivity assay (cell), the IgG level of the KRG group significantly increased but there was no significant difference in the IgG of the CRG group. The IgM level of the KRG group significantly increased stimulated with PWM. When it was unstimulated, the level of IgM in KRG and CRG increased together. The IgA level of the KRG group significantly increased when it was stimulated with PWM and unstimulated. Conclusion : According to the above results, oral administration of red ginseng for 3 months is considered useful for immunomodulatory effect, and Korean red ginseng may be superior to Chinese red ginseng in that effect.

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Enhancement of Sensitivity in Interferometric Biosensing by Using a New Biolinker and Prebinding Antibody

  • Park, Jae-Sook;Lim, Sung-Hyun;Sim, Sang-Jun;Chae, Hee-Yeop;Yoon, Hyun-C.;Yang, Sang-Sik;Kim, Byung-Woo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1968-1976
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    • 2006
  • Recombinant E. coli ACV 1003 (recA:: lacZ) was used to measure low concentrations of DNA-damaging chemicals, which produce $\beta$-galactosidase via an SOS regulon system. Very low $\beta$-galactosidase activities of less than 0.01 unit/ml, $\beta$-galactosidase produced through an SOS response corresponding to the 10 ng/ml (ppb) of DNA damaging chemicals in the environment, can be rapidly determined by using an alternative interferometric biosensor with optically flat thin films of porous silicon rather than by the conventional time-consuming Miller's enzyme assay as well as the ELISA method. fu order to enhance the sensitivity in the interferometry, it needs to obtain more uniform distribution and higher biolinking efficiency, whereas interferometric sensing is rapid, cheap, and advantageous in high throughput by using a multiple-well-type chip. In this study, pore size adjusted to 60 nm for the target enzyme $\beta$-galactosidase to be bound on both walls of a Si pore and a calyx crown derivative was apllied as a more efficient biolinker. Furthermore, anti-$\beta$-galactosidase was previously functionalized with the biolinker for the target $\beta$-galactosidase to be specifically bound. When anti-$\beta$-galactosidase was bound to the calyx-crown derivative-linked surface, the effective optical thickness was found to be three times as high as that obtained without using anti-$\beta$-galactosidase. The resolution obtained was very similar to that afforded by the time-consuming ELISA method; however, the reproducibility was still unsatisfactory, below 1 unit $\beta$-galactosidase/ml, owing to the microscopic non-uniform distribution of the pores in the etched silicon surface.