• Title/Summary/Keyword: IgE antibody

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Production of Monoclonal Antibodies Specific to Korean Mistletoe pectin (KML-C) and Their Characterization (한국산 겨우살이 렉틴 (KML-C)에 대한 단일크론항체의 생산과 특성)

  • 윤택준;유영춘;강태봉;김성훈;김갑수
    • YAKHAK HOEJI
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    • v.45 no.2
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    • pp.180-189
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    • 2001
  • We have reported that water-extracted Korean mistletoe (KM-110) had various biological activities such as antitumor and immunomodulatory activity, and the pectin fraction (KML-C) of the extract was one of major factors related to its biological functions. In this paper, we produced murine monoclonal antibody (mAb) against KML-C. The cAbs obtained were largely classified into two groups according to specificity to KML-C and ML-I, a pectin from European mistletoe. One group mAbs (9H7-D10 and 3C2-lH4) strongly reacted with KML-C, but not ML-I. In contrast, another group cAbs (8Bll-2C5, BE12-3E9 and 5E10-Fl) reacted with both KML-C and ML-1. The subisotypes of these mobs were shown to be IgGl (9H7-lD10, 3C2-lH4 and 8Bll-2C5) or IgM (8E12-3E9 and 5E10-Fl). To develop an assay system for determination of the amount of KML-C, we established the sandwich ELISA (enzyme-linked immunosorbent assay) method using these mAbs and horse radish peroxidase (HRP)-labelled cAbs. In various combinations of the cAbs for coated antibody and detection antibody, the sandwich ELISA quantitatively detected KML-C, showing the detection limit ranging from 7-5,000 ng/ml. Especially reproducibility (C.V) of the sandwich ELISA, in which 8E12-3E9 was used for coating antibody and 8Bll-2C5-HRP for detection antibody, was 4.59-5.83 in intra assay, and 3.9-9.4 in inter assay.

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Decreased Serum Immunoglobulin in Recurrent Otitis Media with Effusion (재발성 삼출성 중이염과 혈청 면역글로불린 농도의 저하)

  • Shin, Il-Ho;Park, Dong-Choon;Byun, Jae-Yong;Park, Moon-Seo;Cha, Chang-Il;Yeo, Seung-Geun
    • IMMUNE NETWORK
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    • v.7 no.2
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    • pp.75-79
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    • 2007
  • Background: Defective or immature antibody responses to pathogens in children may explain the increased susceptibility to acute otitis media. However, there is no study in Korea patients whether a correlation exists between otitis media with effusion and the levels of serum immunoglobulins, IgG subclasses, IgA, IgM and IgE. Methods: 45 children with otitis media with effusion more than 4 episodes in 12 months or 3 episodes in 6 months, 62 children with otitis media with effusion less than 3 episodes in 12 months and 102 children for control group took part in the study at the Department of Otorhinolaryngology of the KyungHee University from May 2004 to Feburary 2007. Serum immunoglobulin levels were determined by nephelometry. And then the relationship between otitis media with effusion and serum immunoglobulin level was evaluated. Results: In otitis media prone group, serum IgG1, IgG2, IgG4, and IgA level was lower than those level of control group, it was significantly decreased (p<0.05). In otitis media group, serum IgA, IgE, and IgG4 level was lower than those level of control. But it was not statistically significant (p>0.05). Conclusion: Lower immunoglobulins in children with otitis media with effusion suggest a generalized decreased antibody responses. Lower levels of serum IgG1, IgG2, IgG4, and IgA may be related with chronicity or intractability of otitis media with effusion.

Seroprevalence of Dengue Virus Antibody in Korea (한국인에서 뎅기바이러스 항체의 혈청 유병률 연구)

  • Lee, Ji Hyen;Kim, Han Wool;Kim, Kyung-Hyo
    • Pediatric Infection and Vaccine
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    • v.25 no.3
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    • pp.132-140
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    • 2018
  • Purpose: The number of dengue fever cases is rising due to increasing overseas travel. Vaccination makes severe dengue fever in seronegative individuals after vaccination when they exposure to wild-type dengue virus. We investigated the seroepidemiology of the dengue virus for monitoring of Korean dengue virus immunity and establishing the prevention of dengue infection. Methods: The study was based on 446 residual sera collected from 98 infants (2 months to 1 year old), 152 adolescents (13 to 19 years old), 90 adults (20 to 50 years old), and 106 elderly participants (more than 65 years old) for other studies. Antibody levels for dengue virus immunoglobulin G (IgG) in each age group were measured using an enzyme-linked immunosorbent assay (ELISA). For each dengue virus IgG positive or equivocal result, an IgG ELISA was performed for Japanese encephalitis virus. Results: Of the 446 serum samples, only 1 (0.2%) adolescent had a positive result from the dengue IgG antibody test. In the dengue virus IgG antibody test, 14 (3.1%) samples showed equivocal results (10 adolescents and 4 elderly). In the 1 positive case of dengue virus IgG, the Japanese encephalitis IgG test was also positive. In the 14 equivocal cases of dengue virus IgG, there were 6 positive, 3 equivocal, and 5 negative of Japanese encephalitis IgG. Conclusions: The seroprevalence rate of dengue virus was very low in Koreans. This study provides important data for establishing the policy for preventive measures of dengue fever. It will be necessary to continuously monitor for dengue virus immunity.

Inhibitory Effects of Chicken Egg Yolk Antibody on Infection of Escherichia coli in Macrophage

  • Lee, Jin-Ju;Kim, Dong-Hyeok;Lim, Jeong-Ju;Kim, Dae-Geun;Kim, Gon-Sup;Min, Won-Gi;Lee, Hu-Jang;Rhee, Man-Hee;Chang, Hong-Hee;Kim, Suk
    • Journal of agriculture & life science
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    • v.46 no.2
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    • pp.107-114
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    • 2012
  • The present study evaluated the potential use of immunoglobulin prepared from egg yolk of chickens immunized with Escherichia coli K88 (IgY-Ec) in the control of E. coli K88 infection in RAW 264.7 murine macrophage. The binding activity of IgY-Ec against E. coli K88 surface protein was more specific and increased than control IgY. In infection assay of E. coli in macrophage, the specific IgY-Ec to E. coli K88 remarkably inhibited the phagocytic activity comparing to nonspecific IgY (p<0.001). In adherence assay, bacterial adhesion on macrophage cells was definitely reduced by preincubation of IgY-Ec compared with nonspecific IgY (p<0.05). These findings suggested that IgY-Ec have the protective effects against pathogens and IgY-based diets may have potential benefits for preventing or treating various infections in domestic animals.

Effects of coffee intake on airway hypersensitivity and immunomodulation: an in vivo murine study

  • Ying-Chi Wong;Wen-Cheng Hsu ;Tzee-Chung Wu ;Ching-Feng Huang
    • Nutrition Research and Practice
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    • v.17 no.4
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    • pp.631-640
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    • 2023
  • BACKGROUND/OBJECTIVES: Coffee is a complex chemical mixture, with caffeine being the most well-known bioactive substance. The immunomodulatory and anti-inflammatory properties of coffee and caffeine impact health in various aspects, including the respiratory system. The objective is to investigate the effects of coffee and caffeine on airway hyperresponsiveness and allergic reactions, as well as to analyze and compare associated cytokine profiles. MATERIALS/METHODS: BALB/c mice were intraperitoneally sensitized with ovalbumin (OVA) and given OVA inhalation to induce airway hypersensitivity. Two weeks after sensitization, they were intragastrically gavaged with coffee or caffeine, both containing 0.3125 mg caffeine, daily for 4 weeks. Control mice were fed with double-distilled water. Serum OVA-specific antibody levels were measured beforehand and 5 weeks after the first gavage. Airway hyperresponsiveness was detected by whole body plethysmography after gavage. Cytokine levels of bronchoalveolar lavage and cultured splenocytes were analyzed. RESULTS: Coffee effectively suppressed T helper 2-mediated specific antibody response. Airway responsiveness was reduced in mice treated with either coffee or caffeine. Compared to the control, coffee significantly reduced OVA-specific immunoglobulin (Ig) G, IgG1 and IgE antibody responses (P < 0.05). Caffeine also attenuated specific IgG and IgG1 levels, though IgE level was unaffected. Coffee significantly reduced interleukin (IL)-4 and increased IL-10 concentration in spleen cells and bronchoalveolar lavage fluid (P < 0.05). CONCLUSIONS: Coffee effectively attenuated airway hyperresponsiveness and systemic allergic responses induced by OVA food allergen in mice. As a complex composition of bioactive substances, coffee displayed enhanced immunomodulatory and anti-inflammatory effects than caffeine.

Effect of Ecklonia cava Water Extracts on Inhibition of IgE in Food Allergy Mouse Model (식품 알레르기 마우스 모델에서 감태 물 추출물의 IgE 분비 억제 효과)

  • Song, Eu-Jin;Lee, Chung-Jo;Kim, Koth-Bong-Woo-Ri;Jung, Ji-Yeon;Kwak, Ji-Hee;Choi, Moon-Kyoung;Kim, Min-Ji;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.12
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    • pp.1776-1782
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    • 2010
  • This research was done to verify the effect of Ecklonia cava water extracts (ECWE) on inhibition of allergic reactions using ovalbumin (OVA)-immunized food allergy mouse model. For in vitro test, $10\sim100{\mu}g$/mL of ECWE and OVA were added to splenocytes obtained from OVA-immunized mice. The significant reduction of IgE antibody level in culture supernatants of splenocytes was shown in ECWE adding group at all tested concentrations. In addition, ECWE decreased IL-4 and IFN-$\gamma$ levels in supernatants of splenocytes. To confirm the effect of ECWE in in vivo test, ECWE was injected to peritoneal cavity of OVA-immunized mice. Subsequently, IgE level was measured in serum and cultured supernatants of splenocytes. As a result, the injection of ECWE (5 and 10 mg/kg BW) significantly attenuated the secretion of IgE antibody in both serum and splenocytes. In conclusion, the present study indicates that ECWE could suppress in a food allergy mouse model through the inhibition of IgE secretion.

Effects of antii-IgE mAb on serum IgE, $Fc{\varepsilon}RII/CD23$ expression on splenic B cells and worm burden in mice infected with Paragonimus westermani (폐흡충 감염 마우스에 있어 Anti-lgE 단일크론 항체 처치시 혈청내 총 IgE, 비장 B 세포표면의 $Fc{\varepsilon}RII/CD23$ 발현 및 충체수에 미치는 영향)

  • 신명헌;민홍기
    • Parasites, Hosts and Diseases
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    • v.35 no.1
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    • pp.47-54
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    • 1997
  • It is generally accepted that parasite-specific IgE plays a crucial role in host defense against helminthic parasites. However, the role of high levels of nonspecific IgE in helminthic infections is still controversial. To investigate the role of nonspecific IgE in primary infections with P. westemani the effect of anti-lgE mAb treatment on serum IgE, $Fc{\varepsilon}RII/CD23$ expression and worm burden in Parcgonimus-infected mice were examined. In mice treated with anti-lgE antibody, the total IgE levels were not detectable ($1{\;}{\mu\textrm{g}/ml}$) throughout the experiment compared with untreated infected mice. The mean percentages of $Fc{\varepsilon}RII/CD23$ positive splenic B cells in anti-lgE treated mice (ridge: 20.3 - 30.5) were also decreased throughout the experiment compared with untreated infected mice (range: 35.7-44.4). Reduction of the total IgE and expression of $Fc{\varepsilon}RII/CD23$ on splenic B cells resulted in decreased worm burden six weeks post infection. These results suggest that high levels of nonspecific IgE in mice with primary infections of P. westemnni play a harmful, rather than beneficial, role for the host, perhaps by interfering with CD23-dependent cellular pathways.

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Detection of Serum IgE Specific to Mite Allergens by Immuno-PCR

  • Lee, Kyung-Woo;Hur, Byung-Ung;Chua, Kaw-Yan;Kuo, I-Chun;Song, Suk-Yoon;Cha, Sang-Hoon
    • IMMUNE NETWORK
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    • v.8 no.3
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    • pp.82-89
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    • 2008
  • Background: Although a skin test is the primary option for detecting allergen-specific IgE in clinics, the serum IgE immunoassay is also important because it allows for the diagnosis of allergy without any accompanying adverse effect on the patient. However, the low detection limit of IgE levels by immunoassay may restrict the use of the method in some occasions, and improving its sensitivity would thus have a significant implication in allergy-immunology clinics. Methods: In this study, we attempted to detect specific serum IgE by using immuno-polymerase chain reaction (IPCR) which combines the antigen-antibody specificity of enzyme-linked immunosorbent assays (ELISAs) with the amplification power of PCR. Results: Our results demonstrated that Blo t5-specific serum IgE can be detected by IPCR with a 100-fold higher sensitivity than ELISA, and cross-reactivity of serum IgE to other mite allergens is able to be analyzed by using only $0.3{\mu}l$ of serum sample. Use of real-time IPCR seemed to permit more convenient determination of specific serum IgE as well. Conclusion: We believe that IPCR can serve as a valuable tool in determining specific serum IgE, especially when the amount of serum sample is limited.

Differentiation of Entomoeba histolyticn and Entcmoeba dispor in cyst-passers by immunoblot (면역이적법을 이용한 아질아메바와 동형아메바의 감별진단)

  • 이미정;홍성태
    • Parasites, Hosts and Diseases
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    • v.34 no.4
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    • pp.247-254
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    • 1996
  • Differentiation of invasive strains of Entamoebn histolytica according to their pathogenicity has been a topic of long debate, but now the pathogenic species only is regarded as E. histolytica while the non-pathogenic species is E. dispar. The present study applied immunoblot to differentiale infections of the two species among microscopically- detected cyst-passers in Korea. The crude extract of 5. histolyticn separated in 5-20% gradient gels, revealed many fractions of 94. 81. 71, 50. 44, 38.5. 37.5, 29, 19. and 18 kDa when the cysteine proteinase inhibitor. E64, was supplemented. The serum IgG antibody of 3 proven E. histolytirc cases reacted loth the antigenic fractions of 117. 110. 99.68,66,60.54.52, 46. and 45 kDa. Sera of PCR confirmed 3 cases of E. disper reacted only to the 117 kDa fraction or the E. histolytica crude extract which was regarded as non specific. To the antitigen of monoxenic E. dispar. sera or E. dispar and E. histolytica cases showed the same immunoblot reactions. The serum IgG antibody reacted with several antigenic fractions of both E. histolytica and E. dispar. but IgM and IgE antibodies showed no reaction to either antigen. Sera of 24 symptomless amebic cyst-passers were screened with the E. histolytica alltigen; two were found to be infected by E. histolytica and 22 were by E. dispar. The present findings suggest that in Korea most of asymptomatic cyst passers of E. histolytica are carriers of E. dispar. Immunoblot using E. histolytica antigen is a good technique for the differentiation of E. histolytica and E. dispar infections.

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Methods for rapid identification of a functional single-chain variable fragment using alkaline phosphatase fusion

  • Lee, Kyung-Woo;Hur, Byung-Ung;Song, Suk-Yoon;Choi, Hyo-Jung;Shin, Sang-Hoon;Cha, Sang-Hoon
    • BMB Reports
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    • v.42 no.11
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    • pp.731-736
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    • 2009
  • The generation of functional recombinant antibodies from hybridomas is necessary for antibody engineering. However, this is not easily accomplished due to high levels of aberrant heavy and light chain mRNAs, which require a highly selective technology that has proven complicated and difficult to operate. Herein, we attempt to use an alkaline phosphate (AP)-fused form of single-chain variable fragment (scFv) for the simple identification of a hybridoma-derived, functional recombinant antibody. As a representative example, we cloned the scFv gene from a hybridoma-producing mouse IgG against branched-chain keto acid dehydrogenase complex-E2 (BCKD-E2) into an expression vector containing an in-frame phoA gene. Functional recombinant antibodies were easily identified by conventional enzyme-linked immunosorbent assay (ELISA) by employing scFv-AP fusion protein, which also readily serves as a valuable immuno-detective reagent.