• Title/Summary/Keyword: immunize

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The Immunogenic Peptide for Th1 Development and Its Adjuvant Activity

  • Takatsu, Kiyoshi
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.171-172
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    • 2002
  • Immune responses to infectious microbes and foreign antigens are regulated by a series of interactions among T cells, B cells, and antigen-presenting cells (APCs) such as macrophages (M$\square$) and dendritic cells (DCs). The inverse relationship between antibody production and cell-mediated immune responses such as delayed type hypersensitivity (DTH) was experimentally manipulated by varying the dose, route of administration, and form of antigen used to immunize animals. (omitted)

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High-level Expression, Polyclonal Antibody Preparation and Bioinformatics Analysis of Bombyx mori Nucleopolyhedrovirus orf47 Encodes Protein

  • Wu, Chao;Guo, Zhongjian;Chen, Keping;Shen, Hongxing
    • International Journal of Industrial Entomology and Biomaterials
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    • v.16 no.2
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    • pp.87-92
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    • 2008
  • Bombyx mori nucleopolyhedrovirus (BmNPV) orf47 gene was characterized for the first time. The coding sequence of Bm47 was amplified and subcloned into the prokaryotic expression vector pET-30a(+) in order to produce His-tagged fusion protein in the BL21 (DE3) cells. The His-Bm47 fusion protein was expressed efficiently after induction with IPTG. The purified fusion protein was used to immunize New Zealand white rabbits to prepare polyclonal antibody. As the genome of BmNPV is available in GenBank and the EST database of BmNPV is expanding, identification of novel genes of BmNPV was conceivable by data-mining techniques and bioinformatics tools. Structural bioinformatics approach to analyze the properties of Bm47 encodes protein.

Immunolocalization of the 150 kDa protein in cyst fluid of Taenia solium metacestodes

  • Yang, Hyun-Jong;Chung, Young-Bae
    • Parasites, Hosts and Diseases
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    • v.42 no.2
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    • pp.81-84
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    • 2004
  • The 150 kDa protein of cyst fluid (CF) of Taenia solium metacestodes was purified by ammonium sulfate fractionation and Superose 6 HR gel filtration chromatography. The purified protein consisted of three subunits (15, 10 and 7 kDa proteins), which were analyzed with the use of a 7.5-15% gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis (SOS-PAGE). Immunofluorescence study was carried out by using immunize specific polyclonal antibody. Positive reactions were noticed at bladder walls, calcareous corpuscles, granules of cyst fluid and some host tissue surrounding the bladder wall of the metacestodes. These results suggest that the 150 kDa protein was secreted into host tissues, inducing immune responses in the host, and it may play important roles in the cellular physiology of the parasites.

A Novel EST with Trench Electrode to Immunize Snab-back Effect and to Obtain High Blocking Voltage

  • Kang, Ey-Goo;Sung, Man-Young
    • Transactions on Electrical and Electronic Materials
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    • v.2 no.3
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    • pp.33-37
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    • 2001
  • A vertical trench electrode type EST has been proposed in this paper. The proposed device considerably improves snapback which leads to a lot of problems of device applications. In this paper, the vertical dual gate Emitter Switched Thyristor (EST) with trench electrode has been proposed for improving snab-back effect. It is observed that the forward blocking voltage of the proposed device is 745V. The conventional EST of the same size were no more than 633V. Because the proposed device was constructed of trench-type electrodes, the electric field moved toward trench-oxide layer, and the punch through breakdown of the proposed EST is occurred at latest.

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Production of Monoclonal Antibodies by Hybridomas Sensitized to Sporozoites of Cryptosporidium parvum (Cryptosporidium parvum Sporozoites 에 감작된 Hybridomas 에서의 Monoclonal Antibody 생산)

  • Cho, Myung-Hwan
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.494-498
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    • 1989
  • Hybridoma cell lines, which secrete monoclonal antibodies (mAbs) against the surface antigens of Cryptosporidium parvum Sporozoites, were produced by fusing spleen cells of C. parvum Sporozoite-immunized mice with P3-X63-Ag8 myeloma cells. Two cloned antibody-secreting cell lines, Kor1 and Ea2, were established and produced IgG1 and IgG2a antibodies, respectively. Percoll-purified sporozoites were solubilized and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Western blot assay demonstrates that an antigen of 20-kDa was bound by monoclonals. By indirect immunofluorescence microscopy, mAb exhibited uniform binding to the sporozoite surface.

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Molecular identification of the vaccine strain from the inactivated bovine viral diarrhea virus (BVDV) vaccines

  • Yang, Dong-Kun;Kim, Ha-Hyun;Cho, Soo-Dong;Choi, Sung-Suk;Kim, Jae-Jo;Song, Jae-Young
    • Korean Journal of Veterinary Service
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    • v.36 no.1
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    • pp.1-6
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    • 2013
  • Since the 1980's, several kinds of inactivated bovine viral diarrhea virus (BVDV) vaccines have been used to immunize domestic animals such as cattle and goat in Korea. Immunogenicity of the BVDV vaccines has been checked by the Korean Veterinary Authority using laboratory animals. In this study, we applied a molecular method to investigate the genetic characterization of the BVDV genes in six commercial inactivated BVDV vaccines, and determined the efficiency of two extraction reagents (i.e., sodium citrate or isopropyl myristate) to separate the vaccine antigens from the antigen/adjuvant complexes. Six partial non-coding regions (288 bp) were successfully amplified with specific primer sets, which demonstrated that sodium citrate is more efficient in extracting viral RNA from inactivated gel vaccines than isopropyl myristae. In addition, we identified the virus strains from the vaccines by analyzing the nucleotide sequences of the 5' non-coding region (NCR) of BVDV. The nucleotide similarity of the partial 5' NCR ranged from 95.1 to 100% among BVDV vaccine strains, respectively, indicating that a few manufacturers used different BVDV strains to produce their vaccines.

Comparison of Properties of Polyclonal Anti-N-Acetylchitooligosaccharides and Anti-Chitooligosaccharides Antibodies Produced for ELISA

  • Shim, Youn-Young;Shon, Dong-Hwa;Kwak, Bo-Yeon;Yu, Jae-Hoon;Chee, Kew-Mahn
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.686-692
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    • 2004
  • To develop the enzyme-linked immunosorbent assay (ELISA) for the analysis of N-acetylehitooligosaccharides (NACOS) and chitooligosaccharides (COS), specific antibodies (Abs) were produced, and their properties were compared. N-acetylehitohexaose (NACOS6), chitohexaose (COS6), and COS mixture (COSM) conjugated to bovine serum albumin (BSA) were used to immunize rabbits. By the use of specific Abs and NACOS6-horseradish peroxidase (HRP), COS6-HRP, and COSM-HRP conjugates, competitive direct ELISA (cdELISA) was developed. The detection limits of NACOS6 by the anti-NACOS6 Ab and COS6 by the anti-COS6 and the anti-COSM Abs in the cdELISAs were about 0.2, 2, ana 2 ng/ml (ppb), respectively. In the cdELISA, the anti-NACOS6 Ab was found to recognize NACOS3-NACOS6, but not N-acetyl-D-glucosamine (GlcNAc), NACOS2, and COSs; the anti-COS6 Ab recognized COS2-COS6 and COSM, but not glucosamine (GlcN) and NACOSs. The recognition pattern of the anti-COSM Ab was almost the same as that of the anti- COS6 Ab, except that the former recognized COS2 and COS3 slightly better than the latter.

Induction of Immunity Against Hepatitis B Virus Surface Antigen by Intranasal DNA Vaccination Using a Cationic Emulsion as a Mucosal Gene Carrier

  • Kim, Tae Woo;Chung, Hesson;Kwon, Ick Chan;Sung, Ha Chin;Kang, Tae Heung;Han, Hee Dong;Jeong, Seo Young
    • Molecules and Cells
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    • v.22 no.2
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    • pp.175-181
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    • 2006
  • Delivery of DNA vaccines to airway mucosa would be an ideal method for mucosal immunization. However, there have been few reports of a suitable gene delivery system. In this study we used a cationic emulsion to immunize mice via the intranasal route with pCMV-S coding for Hepatitis B virus surface antigen (HBsAg). Complexing pCMV-S with a cationic emulsion dramatically enhanced HBsAg expression in both nasal tissue and lung, and was associated with increases in the levels of HBs-specific Abs in serum and mucosal fluids, of cytotoxic T lymphocytes (CTL) in the spleen and cervical and iliac lymph nodes, and of delayed-type hypersensitivity (DTH) against HBsAg. In contrast, very weak humoral and cellular immunities were observed following immunization with naked DNA. In support of these observations, a higher proliferative response of spleenocytes was detected in the group immunized with the emulsion/pCMV-S complex than in the group immunized with naked pCMV-S. These findings may facilitate development of an emulsion-mediated gene vaccination technique for use against intracellular pathogens that invade mucosal surfaces.

Production of Specific Egg Yolk Antibodies in Chicken against Recombinant Fowl Adenovirus Fiber 2 Protein (재조합 가금 아데노바이러스 Fiber 2 단백질을 이용한 특이 난황 항체 생산)

  • Jung, Kyung Min;Lee, Seong;Kim, Jung Woo
    • Korean Journal of Poultry Science
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    • v.41 no.1
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    • pp.15-20
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    • 2014
  • Fowl adenovirus (FAV) is an important cause of several diseases, which result in considerable economic losses to the poultry farm. An outer capsid protein of FAV, fiber 2 is essential for virus growth, assembly or spread. This study was performed to produce about 22 kDa of recombinant fiber 2 protein and to immunize in laying hens to acquire the specific IgY antibody against the recombinant fiber 2. Laying hens were immunized with the recombinant fiber 2 intramuscularly in the breast muscle by injection 4 times at intervals of three weeks. At 12 weeks, serum- and egg yolk-antibody titers of hens against fiber 2 were increased up to 430,000 and 414,000, respectively. The recombinant fiber 2 could be recognized be the anti-His monoclonal antibody. Anti-fiber 2-IgY antibody could recognize the fiber 2 specifically in western blot analysis. These results suggested that the recombinant fiber 2 antigen could be used as an immunogen to elicit IgY antibody against fiber 2 and the anti-fiber 2-IgY could neutralize fowl adenovirus fiber 2 effectively.

Defining B Cell Epitopes of Ovalbumin for the C57BL/6 Mice Immunized with Recombinant Mycobacterium smegmatis

  • Kim, Hyo-Joon;Lee, Yang-Min;Hwang, Joon-Sung;Won, Ho-Shik;Kim, Bok-Hwan
    • BMB Reports
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    • v.32 no.5
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    • pp.461-467
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    • 1999
  • Recombinant Mycobacterium smegmatis expressing ovalbumin was used to immunize C57BL/6(H-$2^b$) mice, and the humoral immunity against recombinant ovalbumin was analyzed. Antibodies were purified by denatured ovalbumin-conjugated affinity chromatography. The epitopes of the antibodies were screened with a random peptide library displayed on the tip of fUSE5 filamentous phage pIII minor coat proteins. Two peptides, IRLADR and SPGAEV, were selected predominantly by the recognition of purified antibodies using biopanning methods. The composition of the peptide sequence with the primary structure of OVA revealed that the peptide sequence analogizes to INEAGR, part of the $^{323}ISQAVHAAHAEINEAGR^{339}$ sequence previously reported as the antigenic determinant for murine Band also Th cell epitopes (I-$A^d$ binding). Also, the structures of these mimotopes obtained from restrained molecular dynamic computations resulted in the formation of a $\beta$-turn proven to be a secondary structure of the parent peptide within the ovalbumin molecule, enabling us to confirm the structural similarity. This study demonstrates that immunization with recombinant M. smegmatis can generate neutralizing antibodies identical with those induced by the administration of natural antigenic proteins and supports the potential use of mycobacteria as vaccine delivery vehicles.

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