• Title/Summary/Keyword: antigen analysis

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Production, Characterization, and Variable Region Analysis of Monoclonal Antibodies Specific for Hepatitis B Virus S Antigen (Hepatitis B Virus의 S항원에 특이적인 단세포군 항체 생산, 특성 연구 및 가변지역유전자 분석)

  • Song, Moo-Young;Kim, Chang-Seok;Park, Sang-Koo;Lee, Jae-Sun;Yoo, Tae-Hyoung;Ko, In-Young
    • IMMUNE NETWORK
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    • v.3 no.4
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    • pp.281-286
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    • 2003
  • Background: Hepatitis B virus (HBV) infection is one of the worldwide public health problem affecting about 300 million people. The envelope protein of HBV consists of three components known as preS1, preS2, and S antigen. According to the recent study, anti-HBs Ab showed effective neutralization ability against HBV from chronic hepatitis B and liver transplant patients, suggesting the possible development of therapeutic antibody. Methods: Spleen cells immunized with S antigen of HBV were fused with myeloma cell line to obtain HBsAg specific monoclonal antibodies. High affinity antibodies against HBsAg (adr, ad and ay type) were selected by competitive ELISA method. Nucleotide sequence of the variable regions of monoclonal antibodies was analyzed by RT-PCR followed by conventional sequencing method. Results: We produced 14 murine monoclonal antibodies which recognize S antigen of HBV. Two of them, A9-11 and C6-9 showed the highest affinity. The sequence analysis of A9-11 revealed that variable regions of the heavy chain and light chains are members of mouse heavy chain I (B) and light chain lambda 1, respectively. Likewise, the sequence analysis of C6-9 revealed that variable regions of the heavy chain and light chains are members of mouse heavy chain II (B) and light chain kappa 1, respectively. Neutralization assay showed that A9-11 and C6-9 effectively neutralize the HBV infection. Conclusion: These results suggest that A9-11 and C6-9 mouse monoclonal antibodies can be used for the development of therapeutic antibody for HBV infection.

An Analysis of Prognostic Factors in the Uterine Cervical Cancer Patients (자궁경부암 환자의 예후인자에 관한 분석)

  • Yang, Dae-Sik;Yoon, Won-Sub;Kim, Tae-Hyun;Kim, Chul-Yong;Choi, Myung-Sun
    • Radiation Oncology Journal
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    • v.18 no.4
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    • pp.300-308
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    • 2000
  • Purpose :The aim of this study is to analysis of suwival and recurrence rates of the uterine cervical carcinoma patients whom received the radiation therapy respectively. The prognostic factors, such as Papanicolaou (Pap) smear, carcinoembriogenic antigen (CEA) and squamous cell carcinoma (SCC) antigen has been studied. Methods and Materials : From January 1981 to December 1998, eight-hundred twenty-seven uterine carvical cancer patients were treat with radiation therapy. All of the patients were divided into two groups : the radiation therapy only (S2l patients) group and the postoperative radiation therapy (326 patients) group. The age, treatment modality, clinical stage, histopathology, recurrence, follow-up Pap smears, CEA and SCC antigen were used as parameters for the evaluation. The prognostic factors such as survival and recurrence rates were peformed with the Kaplan-Meier method and the Cox hazard model, respectively. Median rollow-up was 38.6 months. Results :On the radiation therapy only group, 314 patients (60$\%$) achieved complete response (CR), 47 patients (9$\%$) showed local recurrence (LR), 78 patients (15$\%$) developed distant metastasis (DM). On the Postoperative radiation therapy group, showed 276 Patients (85$\%$) CR, 8 Patients (2$\%$) LR, 37 Patients (11$\%$) DM. The 5-year survival and recurrence rates was evaluated for all parameters. The statistically significant factors for the survival rate in univariate analysis were clinical stage (p=0.0001), treatment modality (p=0.0010), recurrence (p=0.0001), Pap smear (p=0.0329), CEA (p=0.0001) and SCC antigen (p=0.0001). Conclusion: This study indicated that after treatment, the follow-up studies of Pap smear, CEA and SCC antigen were significant parameter and prediction factors for the survival and recurrence of the uterine cervical carcinoma.

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Bordetella bronchiseptica is a potent and safe adjuvant that enhances the antigen-presenting capability of dendritic cells

  • Lee, You-Jeong;Han, Yong;Joo, Hong-Gu
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.1
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    • pp.47-52
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    • 2020
  • We previously demonstrated that Bordetella bronchiseptica (B. bronchiseptica) antigen (Ag) enhances the Mycoplasma hyopneumoniae Ag-specific immune response. The focus of this study was whether acellular bacterin of B. bronchiseptica could be used as an adjuvant to increase antigen-presenting capability of dendritic cells (DCs) by increasing the level of activation. The metabolic activity of DCs was increased by B. bronchiseptica, similar to lipopolysaccharide (LPS). Flow cytometry analysis revealed that B. bronchiseptica increases the expression of major histocompatibility complex class-2, cluster of differentiation (CD)40, CD54, and CD86 which are closely related to DC-mediated immune responses. B. bronchiseptica enhanced the production of cytokines related to adaptive immune responses. Furthermore, the survival rate of B. bronchiseptica-injected groups was 100% at 15 and 20 mg/kg doses, whereas that of LPS-injected groups was only 20%, 0% at 15 and 20 mg/kg doses respectively, and so B. bronchiseptica is likely to be safer than LPS. Taken together, these results indicate that B. bronchiseptica can be used as an adjuvant to enhance the antigen-presenting capability of DCs. B. bronchiseptica is a candidate for producing vaccines, especially in case of DC-mediating efficacy and safety demands. This study provides researchers and clinicians with valuable information regarding the usage of B. bronchiseptica as a safe bacteria-derived immunostimulating agent for developing efficient vaccines.

Differential Activation of T Cells by T-Cell Receptor Ligand Analogs

  • Choi, Yun-Hi;Suh, Yu-Jin;Kim, Kil-Hyoun
    • BMB Reports
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    • v.30 no.6
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    • pp.415-420
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    • 1997
  • Although $CD4^+$ T cell responses to protein-derived antigen have well been understood, the epitopes recognized by hapten-specific $CD4^+$ T cells have not been fully defined. In this study, we characterized the response of a T cell hybridoma (5Di0.1B8) which is specific for a hapten. N-hydroxysuccinimidyl-4-azidobenzoate (HSAB) restricted by MHC class II $I-A^d$. Using three different antigen presenting cells (APCs) expressing $I-A^d$, the role of class II MHC proteins in haptenic antigen presentation and subsequent activation of 5D10.1B8 has been examined. Activation of 5D10.1B8 T cells by HSAB analogs was also performed. Our results show that each APC activated T cells differentially and that interleukin-2 (IL-2) augmented antigen-presenting ability of all the APCs, suggesting that increased expression of class II MHC protein by IL-2 played an important role in HSAB presentation and T cell activation. Finally, early T cell receptor-dependent signals induced by HSAB or its analogs were examined by phosphotyrosine immunoblot analysis, and showed that tyrosine phosphorylation level of a 18-20 kD protein increased upon stimulation.

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Serodiagnosis of Echinococcosis by ELISA Using Cystic Fluid from Uzbekistan Sheep

  • Jin, Yan;Anvarov, Khikmat;Khajibaev, Abdukhakim;Hong, Samin;Hong, Sung-Tae
    • Parasites, Hosts and Diseases
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    • v.51 no.3
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    • pp.313-317
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    • 2013
  • According to increase of travel, the cases of imported echinococcosis have been increasing in Korea. The present study was undertaken to develop a serodiagnostic system for echinococcosis in Korea. For diagnosis of echinococcosis, the fluid of Echinococcus granulosus hydatid cysts was collected from naturally infected sheep in Uzbekistan. Also serum samples of infected patients who were surgically confirmed were collected in a hospital in Tashkent, Uzbekistan. According to the absorbance of 59 echinococcosis positive and 39 negative control serum samples, the cut-off value was determined as 0.27. The sensitivity and specificity of ELISA with hydatid fluid antigen were 91.5% and 96%, respectively. The antigen cross-reacted with the serum of some cysticercosis or clonorchiasis patients. However, immunoblot analysis on the cystic fluid recognized antigenic proteins of 7-, 16-, and 24-kDa bands in their dominant protein quantity and strong blotting reactivity. In conclusion, the present ELISA system using hydatid cyst fluid antigen from Uzbekistan sheep is sensitive and specific for diagnosis of echinococcosis cases.

Immunological Identification of Thiobacillus ferro-oxidans and Thiobacillus thiooxidans (Thiobacillus ferrooxidans와 Thiobacillus thiooxidans의 면역학적 동정)

  • 이강순;장정순;이강석
    • Korean Journal of Microbiology
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    • v.16 no.1
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    • pp.30-40
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    • 1978
  • Detergent soluble fractions were obtained from T. ferrooxidans ATCC 13598 and the T. thiooxidans ATCC 8085 which were treated with 3% of Tween 20. The detergent soluble antigen(crude antigen) fractions of the T.ferrooxidans and the T.thiooxidans were subjected to hydroxyapatite. In the case of T.thiooxidans, further purification was carried out on the DEAE-cellulose column chromatography. The antigen fractions, such as the hydroxyapatite peak-1(Tf, HA-1) and peak-2 from T.ferrooxidans(Tf. HA-2) and hydroxyapatite peak-1(Tt, HA-1), DEAE-cellulose peak-1(Tt, DP-1) and peak-2(Tt, DP-2) from T. thiooxidans wre compared each other with the homologous and the heterologous and the heterologous antisera against to the Thiobacillus species. The hydroxyapatite peak-2 fraction from the T.ferrooxidans(Tf, HA-2) and DEAE-cellulose peak-2 fraction from the T.thiooxidans(Tt, DP-2) were represented the type-specific immuno-reactivities between the T.ferrooxidans and the T.thiooxidans on the several sets of double gel diffusioin analysis. The type-specific antigen fractions from both of the baceteria were mainly composed of protein with entierly different electrophoretic mobility on the SDS-polyacrylamide gel electrophoresis. However, the PAS positive banding patterns on the electrophorogram showed wide range of common antigenic properties in the T. ferrooxidans and the T.thiooxidans, respectively.

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Role of CAGE, a Novel Cancer/Testis Antigen, in Various Cellular Processes, Including Tumorigenesis, Cytolytic T Lymphocyte Induction, and Cell Motility

  • Kim, Young-Mi;Jeoung, Doo-Il
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.600-610
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    • 2008
  • A cancer-associated antigen gene (CAGE) was identified by serological analysis of a recombinant cDNA expression library (SEREX). The gene was identified by screening cDNA expression libraries of human testis and gastric cancer cell lines with sera from patients with gastric cancer. CAGE was found to contain a D-E-A-D box domain and encodes a putative protein of 630 amino acids with possible helicase activity. The CAGE gene is widely expressed in various cancer tissues and cancer cell lines. Demethylation plays a role in the activation of CAGE in certain cancer cell lines where the gene is not expressed. The functional roles of CAGE in tumorigenesis, the molecular mechanisms of CAGE expression, and cell motility are also discussed.

The cancer/testis antigen CAGE induces MMP-2 through the activation of NF-κB and AP-1

  • Kim, Young-Mi;Jeoung, Doo-Il
    • BMB Reports
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    • v.42 no.11
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    • pp.758-763
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    • 2009
  • Cancer-associated antigen (CAGE) induces the expression of matrix metalloproteinase-2 (MMP-2) by activating Akt, which in turn interacts with inhibitory kappa kinase $\beta$ ($I{\kappa}K{\beta}$) to activate nuclear factor ${\kappa}B$ (NF-${\kappa}B$). Akt and p38 mitogen activated protein kinase (p38 MAPK) are necessary for CAGE-mediated induction of the AP-1 subunit JunB, whereas extracellular regulated kinase (ERK) is necessary for the induction of fos-related antigen-1 (Fra-1). Induction of MMP-2 by CAGE requires activator of protein-1 (AP-1) to be bound. Specific binding of JunB to MMP-2 promoter sequences was shown by chromatin immunoprecipitation (ChIP) analysis.

The Effect of Cyclosporine A on the Expression of the Major Histocompatibility Complex Antigen Class II(MHC II) (Cyclosporine A의 MHC Class II 항원에 대한 억제 효과)

  • 박국양
    • Journal of Chest Surgery
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    • v.28 no.5
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    • pp.443-446
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    • 1995
  • The present study was designed to determine whether cyclosporine A inhibits Major Histocompatibility Complex Class II antigen[MHC II expression in the allograft rat heart myocardium. In this rat allograft study we also tried to elucidate whether CSA inhibits MHC II in a dose dependent way. Hearts were isolated from LBN rats weighing 200-250 grams and heterotopically transplantated into the abdomen of weight-matched ACI rats. The ACI rats were randomly assigned to one of the three experimental groups according to cyclosporine dosage: {1}control [no CSA , n=6 {2}CSA 5 mg/day , n=5 {3}CSA 10 mg/day, n=5. The transplanted hearts were harvested 5 days postoperatively and analyzed. MHC II expression was detected by indirect immunoperoxidase staining and quantified via computer image analysis system. The % positive area reading was obtained in each slide [50 areas per group and compared to other groups. Significant differences were noted between three groups [p<0.05 . We conclude that CSA inhibits MHC II in heterotopically transplanted allograft rat heart in a dose dependent way.

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Analysis of Integrity of Killed Hantavirus Vaccine by Antigen-Capture Reverse Transcriptase PCR

  • HWANG KYUNG-A;JOO YOUNG-RAN;SHIN YOUNG-HAK;PARK KEUN-YONG;NAM JAE-HWAN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1384-1387
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    • 2005
  • Hantavax(R) is one of the killed Hantavirus vaccines, and is commercially available in South Korea. This vaccine was developed by inactivation of virus isolated from infected suckling mouse brain with formalin. Although Hantavax(R) can induce neutralizing antibodies in vaccinees, the strength of this induction and the duration of the humoral immune response are controversial issues. In this study, we studied the native conformation of the killed vaccine by antigen-capture reverse transcriptase polymerase chain reaction with patient and vaccinee sera containing neutralizing antibodies against Hantavirus. The results showed that Hantavax(R) could bind HTNV patient and vaccinee sera like live virus, suggesting that the integrity of the viral epitope is maintained in Hantavax(R) and induces the protective antibodies, even though the virus was inactivated with formalin.