• Title/Summary/Keyword: opsonization

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Effect of Opsonization of Zymosan on the Chemiluminescent Response of Haemocytes in the Pacific oyster Crassostrea gigas

  • Ahn, Kyoung-Jin;Kim, Ki-Hong
    • Journal of fish pathology
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    • v.14 no.2
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    • pp.103-105
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    • 2001
  • The effect of opsonization of zymosan with serum on the repiratory burst of haemocytes isolated from the Pacific oyster Crassostrea gigas was investigated. The reactive oxygen species (ROS) produced by the respiratory burst of haemocytes in response to the opsonized or unopsonized zymosan were measured using chemiluminescence (CL). The CL values were increased according to the increment of haemocyte number. The degree of CL amplification by increase of haemocytes from $0.1{\times}10^6$ to $0.5{\times}10^6$ cells/ml was 3-5 times, but comparatively low amplification was elicited by increase of haemocytes from $0.5{\times}10^6$ to $1{\times}10^6$ cells/ml. Chemiluminescences produced by the haemocytes in response to the zymosan opsonized with oyster serum were considerably higher than the CL produced by the haemocytes in response to the unopsonized zymosan.

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Antibody Induced by the JY-Pol Pneumococcal Conjugate Protects Mice Against systemic Infection Due to Streptococcus pneumoniae (JY-Pol 접합백신으로 유도된 항페렴구균 항체의 보호효과)

  • Lee, Jue-Hee;Han, Yong-Moon
    • YAKHAK HOEJI
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    • v.48 no.6
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    • pp.369-373
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    • 2004
  • We previously reported that Streptococcus pneumoniae capsule attached to the surface protein (JY-Pol) was protective to systemic pneumococcal infection. The JY -Pol antigen induced IgM, IgG, and IgA in mice and provoked cell-mediated immunity. In this current study, we investigated the effect of anti JY-Pol antiserun and monoclonal antibody C2 (Mab C2) specific for the JY-Pol antigen against the pneumococcal disease. Mice that were given the antiserum survived longer than mice that received antiserum pre-absorbed with S.pneumoniae cells or DPBS as a negative control. Heat-treated anti JY-Pol antiserum resulted in survival rates similar to intact fresh JY-Pol antiserum. Mab C2 isolated from JY-Pol-immunized mice also enhanced resistance of naive mice against the pneumococcal diseaser. This protection by Mab C2 appeared to be mediated by opsonization as determined in a RAW 264.7 monocyte/macrophage cell line. Epitope analysis showed that Mab C2 epitope consisted of glucuronic acid and glucose that blocked the interaction of JY-Pol to the C2. Taken together, these data indicate that the antiserum induced by the JY-Pol, a naturally pneumococcal conjugate formula, mediated the protection by passive transfer, which was confirmed by protective effect of Mab C2.

Functional Immunity to Cross-Reactive Serotype 6A Induced by Serotype 6B in Pneumococcal Polysaccharide Vaccine (폐구균 다당질 백신 내 혈청형 6B에 의해 유도되는 교차 반응 혈청형 6A에 대한 기능적 면역)

  • Kim, Kyung Hyo
    • Clinical and Experimental Pediatrics
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    • v.48 no.5
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    • pp.506-511
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    • 2005
  • Purpose : Streptococcus pneumoniae serotype 6B and 6A are important pathogens in pneumococcal infections. It is commonly assumed that the 6B vaccines elicit antibodies cross-reacting with the 6A serotype and the cross-reactive antibodies protect against infections of 6A. To examine this assumption, we measured the opsonophagocytic capacity to serotype 6A and 6B in adults. Methods : Twenty-four adults were immunized with pneumococcal PS vaccine that contains 6B PS. Their preimmune and postimmune sera were studied for the capacity to opsonize 6B and 6A serotypes with opsonophagocytic killing assay. Results : Opsonization titers to 6B were significantly higher than those to 6A in preimmune and postimmune sera. Because significant increasesof opsonization titers were observed in adults with polysaccharide vaccines for 6A(cross-reactive) serotype as well as for 6B(vaccine) serotype, 6B PS in vaccine elicited cross-protective antibodies to 6A, but not in all cases. One adult did not have detectable levels of opsonization titers to 6A after immunization. Conclusion : Although 6B PS in pneumococcal PS vaccine elicits antibodies cross-reacting with 6A serotype in some adults, it may not occur always. This study should be extended to other age groups such as children and elderly people. The presence of the cross-protection should be directly determined in clinical trials of the pneumococcal vaccines as well as during the postlicensure monitoring surveys by serotyping the clinical isolates of pneumococci.

Formulation and Characterization of Antigen-loaded PLGA Nanoparticles for Efficient Cross-priming of the Antigen

  • Lee, Young-Ran;Lee, Young-Hee;Im, Sun-A;Kim, Kyung-Jae;Lee, Chong-Kil
    • IMMUNE NETWORK
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    • v.11 no.3
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    • pp.163-168
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    • 2011
  • Background: Nanoparticles (NPs) prepared from biodegradable polymers, such as poly (D,L-lactic acid-co-glycolic acid) (PLGA), have been studied as vehicles for the delivery of antigens to phagocytes. This paper describes the preparation of antigen-loaded PLGA-NPs for efficient cross-priming. Methods: NPs containing a similar amount of ovalbumin (OVA) but different sizes were produced using a micromixer-based W/O/W solvent evaporation procedure, and the efficiency of the NPs to induce the cross-presentation of OVA peptides were examined in dendritic cells (DCs). Cellular uptake and biodistribution studies were performed using fluorescein isothiocyanate (FITC)-loaded NPs in mice. Results: The NPs in the range of $1.1{\sim}1.4{\mu}m$ in size were the most and almost equally efficient in inducing the cross-presentation of OVA peptides via $H-2K^b$ molecules. Cellular uptake and biodistribution studies showed that opsonization of the NPs with mouse IgG greatly increased the percentage of FITC-positive cells in the spleen and lymph nodes. The major cell type of FITC-positive cells in the spleen was macrophages, whereas that of lymph nodes was DCs. Conclusion: These results show that IgG-opsonized PLGA-NPs with a mean size of $1.1{\mu}m$ would be the choice of biodegradable carriers for the targeted-delivery of protein antigens for cross-priming in vivo.

Flow cytometry As a Tool for Monitoring Immune Parameters of the Manila clam Ruditapes philippinarum

  • Park, Kyung-Il;Philippe Soudant;Park, Kwang-Sik
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2003.05a
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    • pp.369-369
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    • 2003
  • Hemocytes in marine bivalves play important immunological roles in discrimination, opsonization and phagocytosis of foreign materials as a defense mechanism. In this study we report the flow cytometric implications to investigate the immune parameters such as the compositional and the functional characteristics of hemocytes isolated from the Manila clams, Ruditapes philippinarum. Heterogeneity of the hemocytic cell population was determined by the forward scatter (FSC) and side scatter (SSC) cytometric profile which showed three populations: granulocytes, hyalinocytes and small agranular cells. In addition, phagocytosis rate was measured after adding fluorescent-labeled particles. The data were initially analysed for two-parameters: FSC and SSC, then the fluorescent (FL 1) frequency distribution histogram of the hemocyte population was subsequently obtained.

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Pharmacological, Toxicological Studies of Antitumor Polysaccharides Obtained from Ganoderrna lucidurn IY 009 (Ganoderma lucidum IY 009로 부터 분리된 항암성 다당류의 약리 및 독성)

  • Lee, Kweon-Haeng;Lee, Chong-Ock;Lee, June-Woo;Jeong, Hoon;Han, Man-Deuk;Jeong, June-Ho;Oh, Doo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.22 no.2
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    • pp.182-189
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    • 1994
  • The highest antitumor activity was observed in water soluble AS fraction of the Ganoderma lucidum IY 009. AS fraction did not show any cytotoxicity on sarcoma 180 cell but stimulated antibody production, opsonization of macrophage in ICR mouse and superoxide ion production from isolated macrophage. AS fraction activated complement C3 in human serum, and their antitumor activity was inhibited by EDTA, a chelator of cation related complementary activation. AS fraction exerted om prolong of life span and ingibition of tumor growth in the leukemia P388 or L1210 transplanted inbreed mouse,k BDF1 but krestin did not. AS fraction did not show any serious and lethal effects through oral administration on ICR mouse, and LD$_{50}$ of those was above 2,230 mg/kg.

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Intracellular delivery and anti-tumor activity of polyethyleneglycol liposomes containing cationic lipid (양이온성 지질이 포함된 PEG 리포솜의 세포내 이입 및 항암효력 평가)

  • Jung, Soon-Hwa;Kim, Sung-Kyu;Jung, Suk-Hyun;Seong, Ha-Soo;Cho, Sun-Hang;Shin, Byung-Cheol
    • Journal of Pharmaceutical Investigation
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    • v.38 no.3
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    • pp.163-169
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    • 2008
  • Liposomes are spherical vesicles composed of lipid bilayer membranes. However, the conventional liposomes have been found to be plagued by rapid opsonization and taken up by the reticuloendothelial system (RES), resulting in shortened circulation time and limited intracellular uptake to target cell. In this study, polyethyleneglycol-cationic liposomes (PCL) containing cationic lipid and DSPE-mPEG were prepared by thin film cast-hydration method. The PEG liposomes had approximately $97.0{\pm}1.3\;nm$ of mean particle diameter and $-21.7{\pm}1.2\;mV$ of zeta potential value. PCL had $96.4{\pm}1.8\;nm$ of mean particle diameter and $-8.7{\pm}1.1\;mV$ of zeta potential value with a decrease of about 10 mV compared to the PEG liposomes. Loading of model drug, doxorubicin (DOX), in liposomes were carried out by using remote loading method and the loading efficiency of DOX in liposomes was about $95.0{\pm}1.9%$. Intracellular uptake and cytotoxicity of PCL were higher than that of PEG liposomes to murine B16F10 melanoma cells. In addition, anti-tumor activity of PCL was similar to that of PEG liposomes on growth of A549 human lung carcinoma in BALB/c mice. Consequently, PCL modified with cationic lipid may be applicable as anticancer drug carriers that can increase intracellular uptake and therapeutic efficacy.

Conceptus-derived cytokines interleukin-1β and interferon-γ induce the expression of acute phase protein serum amyloid A3 in endometrial epithelia at the time of conceptus implantation in pigs

  • Soohyung Lee;Inkyun Yoo;Yugyeong Cheon;Hakhyun Ka
    • Animal Bioscience
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    • v.36 no.3
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    • pp.441-450
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    • 2023
  • Objective: Serum amyloid A3 (SAA3), an acute phase response protein, plays important roles in opsonization, antimicrobial activity, chemotactic activity, and immunomodulation, but its expression, regulation, and function at the maternal-conceptus interface in pigs are not fully understood. Therefore, we determined the expression of SAA3 in the endometrium throughout the estrous cycle and at the maternal-conceptus interface during pregnancy. Methods: Endometrial tissues from pigs at various stages of the estrous cycle and pregnancy and with conceptuses derived from somatic cell nuclear transfer (SCNT), conceptus tissues during early pregnancy, and chorioallantoic tissues during mid- to late pregnancy were obtained and the expression of SAA3 was analyzed. The effects of the steroid hormones, interleukin-1β (IL1B), and interferon-γ (IFNG) on the expression of SAA3 were determined in endometrial explant cultures. Results: SAA3 was expressed in the endometrium during the estrous cycle and pregnancy, with the highest level on day 12 of pregnancy. The expression of SAA3 in the endometrium was significantly higher on day 12 of pregnancy than during the estrous cycle. Early-stage conceptuses and chorioallantoic tissues during mid to late pregnancy also expressed SAA3. The expression of SAA3 was primarily localized to luminal epithelial cells in the endometrium. In endometrial explant cultures, the expression of SAA3 was induced by increasing doses of IL1B and IFNG. Furthermore, the expression of SAA3 decreased significantly in the endometria of pigs carrying conceptuses derived from SCNT on day 12 of pregnancy. Conclusion: These results suggest that the expression of SAA3 in the endometrium during the implantation period increases in response to conceptus-derived IL1B and IFNG. The failure of those appropriate interactions between the implanting conceptus and the endometrium leads to dysregulation of endometrial SAA3 expression, which could result in pregnancy failure. In addition, SAA3 could be a specific endometrial epithelial marker for conceptus implantation in pigs.

The Role of Specific IgG in Phagocytosis of Actinobacillus(Haemophilus) Actinomycetemcomitans Y4 by Human Neutrophils (다형핵 백혈구의 Actinobacillus actinomycetemcomitans Y4 균주 탐식시 특이항체의 역할)

  • Kim, Jin-Myung;Chung, Chong-Pyoung;Lee, Young-Hee;Lee, Jin-Yong
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.4
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    • pp.441-446
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    • 1986
  • Previous studies have demonstrated that phagocytosis of encapsulated bacteria needs the opsonization of bacteria with immunoglobulin and complement. Several investigators have studied the role of specific antibody to the bacteria. The purpose of this study is to investigate the role of specific anti-Actinobacillus actinomycetemcomitans Y4($A{\alpha}Y4$) antibody, which was obtained from the immunized rabbit serum for phagocytosis of $A{\alpha}Y4$ by PMNL. For this study, specific and nonspecific IgG were separated from the sera of the rabbits and PMNL were isolated from 15 healthy adults. By an enzyme-linked immunosorbent assay, the results showed that the binding capacity of anti-$A{\alpha}Y4$ IgG to $A{\alpha}Y4$ was much higher than that of nonspecific IgG; 0.75 and 0.14(O.D. at 400nm), respectively. The oxygen consumption of PMNL, phagocytizing $A{\alpha}Y4$ which was opsonized with specific $A{\alpha}Y4$ IgG(37.13 nmol/min/$1{\times}10^7$ PMNL), was significantly higher than that with nonspecific IgG(27.95 nmol/min/$1{\times}10^7$ PMNL, p<0.01). In immunofluorescence microscopic examination, the difference between the numbers of the ingested $A{\alpha}Y4$ opsonized with specific anti-$A{\alpha}Y4$ IgG and nonspecific IgG reached to statistically significant level; $184{\pm}11.4$ and $133.2{\pm}8.3$ per 100 PMNL, p<0.05. These results suggest that specific anti-$A{\alpha}Y4$ IgG has a significant role in PMNL phagocytosis of encapsulated $A{\alpha}Y4$ and also it can be available to adopt this system to develop anti-capsular antibody to $A{\alpha}Y4$ for enhancing and emphasizing the phagocytic activity against this bacterium.

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Preparation of Protein-coated Cationic Liposomes Containing Doxorubicin and Their Binding Property of Blood Plasma Protein (독소루비신을 함유하고 단백질로 수식된 양이온성 리포솜의 제조 및 혈장 단백흡착 특성)

  • Kim, Sung-Kyu;Jung, Soon-Hwa;Jung, Suk-Hyun;Seong, Ha-Soo;Chi, Sang-Cheol;Cho, Sun-Hang;Shin, Byung-Cheol
    • Journal of the Korean Chemical Society
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    • v.52 no.1
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    • pp.57-65
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    • 2008
  • are nanometer or micrometer scale vesicles that can be used as drug delivery carriers. However, plain liposomes are plagued by rapid opsonization, making their circulation time in bloodstream be shortened. In this study, model protein, bovine serum albumin (BSA)-coated liposomes were prepared by coating cationic liposomes with BSA molecules at higher pH than isoelectric point of BSA. The BSA molecules coated on the liposomal surface were denatured by thermal treatment at above 60oC. While both plain and cationic liposomes had about mean particle diameter of 1041 nm, BSA-coated cationic liposomes (BCL) had mean particle diameter of 1091 nm. Encapsulation of model drug, doxorubicin (DOX), in liposomes were carried out by using remote loading method and the loading efficiency of DOX to liposomes was about 90%. The mean particle diameter of BCL did not increase in blood plasma and adsorption of plasma protein was much less than plain or cationic liposomes. These results suggest that BCL can be used as a long-circulating liposomes in bloodstream.