• Title/Summary/Keyword: Antigens

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Antigenic profile and localization of Clonorchis sinensis proteins in the course of infection

  • Hong, Sung-Jong;Kim, Tae-Yun;Song, Kye-Yong;Sohn, Woon-Mok;Kang, Shin-Yong
    • Parasites, Hosts and Diseases
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    • v.39 no.4
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    • pp.307-312
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    • 2001
  • In the course of Clonorchis sinensis infection, antigens presented to the hosts may be in a close relation to growth of the fluke. The antigenic proteins stimulating IgG antibody production were chronologically identified by immunoblot and localized by immunogistochemical staining. In the early stage of infection until 12 weeks post-infection (PI) , antigens were proteins with molecular mass larger than 34 kDa which were derived from the tegument, testes and intrauterine eggs. After 20 weeks PI antigens recognized were 29, 27 and 26 kDa proteins from the intestine, excretory bladder and reproductive organs. It is suggested that the tegumental proteins are the most potent antigens and the excretory-secretory proteins with middle molecular mass of 26-45 kDa contribute to the high level production of antibodies after 20 weeks of the C. sinensis infection.

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Immunohistochemistry for detection of Aujeszky's disease virus antigens: Protein A-gold labeling of ultrathin sections for electron microscopy (오제스키병 바이러스 항원검출을 위한 면역조직화학적 연구 : 전자현미경적 관찰을 위한 초박절편내 protein A-gold labeling)

  • Kim, Soon-bok
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.541-548
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    • 1989
  • The present study was carried out to determine viral antigens and its morphogenesis in the ultrathin frozen and araldite sections of cell cultures infected with ADV by protein A-gold labeling. ADV antigens were labeled with 10nm gold probes, and electron-dense gold particles were mainly present on viral nucleocapsids and viral envelopes. Immunogold labeling in the ultracryosections showed a very low degree of interaction with tissue structures. Immunogold labeling in the ultrathin cryosections can be useful tool for the detection of ADV antigens, and the technique also may provide its great potential for immunocytochemical studies on various virus-host cell Interactions.

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Immunohistochemistry for detection of Aujeszky's disease virus antigens : Immunogold-silver method in tissue sections (오제스키병 바이러스 항원검출을 위한 면역조직화학적 연구 -조직절편내 immunogold-silver법-)

  • Kim, Soon-bok
    • Korean Journal of Veterinary Research
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    • v.28 no.2
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    • pp.365-369
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    • 1988
  • The present study was done to demonstrate ADV antigens in frozen and paraffin sections from ADV-infected pigs and cell cultures by using of the IGS method. Tissue specimens from 3 young pigs infected with ADV-phylaxia strain and of 2 healthy pigs were used. Fibroblastic cells originated from pig brain and BHK cells were grown and confluent monolayers were infected with the virus. Two monoclonal antibodies and a specific hyperimmune serum to ADV were used as the source of primary antibodies for both the IGS and immunoperoxidase methods. Application of the IGS method yielded a black fine granular reaction in positive areas, and the results were superior to those obtained using the immunoperoxidase technique for all cases tested. The IGS method might be useful in the detection of various viral antigens in tissue sections.

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Identification of novel Leishmania major antigens that elicit IgG2a response in resistant and susceptible mice

  • MOHAMMADI Mohammad Reza;ZEINALI Majid;ARDESTANI Sussan K.;KARIMINIA Amina
    • Parasites, Hosts and Diseases
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    • v.44 no.1 s.137
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    • pp.43-48
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    • 2006
  • Experimental murine models with high, intermediate and low levels of genetically based susceptibility to Leishmania major infection reproduce almost entire spectrum of clinical manifestations of the human disease. There are increasing non-comparative studies on immune responses against isolated antigens of L. major in different murine strains. The aim of the present study was to find out whether there is an antigen that can induce protective immune response in resistant and susceptible murine strains. To do that, crude antigenic extract of procyclic and metacyclic promastigotes of L. major was prepared and subjected to SDS-PAGE electrophoresis. Western-blotting was used to search for antigen(s) capable of raising high antibody level of IgG2a versus IgG1 in the sera of both infected resistant and susceptible strains. Two novel antigens from metacyclic promastigotes of L. major (140 and 152 kDa) were potentially able to induce specific dominant IgG2a responses in BALB/c and C57BU6 mice. The 2 antigens also reacted with IgG antibody of cutaneous leishmaniasis patients. We confirm that 140 and 152 kDa proteins of L. major promastigotes are inducing IgG production in mice and humans.

Immunocytochemical identification of Aujeszky's disease viral antigens in nasal and circulating white blood cells from living pigs for early diagnosis (오제스키병의 생체 조기진단을 위한 돼지 비세포 및 순환백혈구내 면역세포화학적 바이러스 항원동정)

  • Moon, Oun-kyong;Cho, Hee-tack;Kim, Soon-bok;Sur, Jung-hyang
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.241-247
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    • 1993
  • Forty day old piglets were intranasally inoculated with 2ml of Aujeszky's disease virus (NYJ-1-87 strain, $10^{7.0}$ $TCID_{50/0.2}ml$), and the viral antigens were detected in nasal and circulating white blood cells for 20 days after inoculation by immunocytochemical method. Antibody titers in the blood were also detected by neutralizing test and Aujeszky's disease serodiagnostic kit(Choong Ang) in this periods. 1. Viral antigens were detected by the immunocytochemical technigue, and positive reactions were observated in nasal cells from the 2nd to the l0th days after inoculation and circulated white blood cells from the 4th to the 12th days after inoculation. 2. In neutralization test antibodies levels showed titers of 2 on the 8th day, 8 on the l5th day, 16 on the 18th day and 32 on the 20th day after inoculation. In serodiagnostic kit test positive reactions were observed after the 15th day after inoculation.

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Western blot diagnosis of vivax malaria with multiple stage-specific antigens of the parasite

  • Son, Eui-Sun;Kim, Tong-Soo;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.171-176
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    • 2001
  • Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), meroxoite surface protein (MSP-1), apical merozoite antigen (AMA- 1), serine repeat antigen (SERA), and exported antigen (EXP- 1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce the expression of GST fusion protein in Escherichia coli by IPTG. The bacterial cell extracts were separated on 10% SDS-PAGE followed by western blot analysis with patient sera which was confirmed by blood smear examination. When applied with patient sera, 147 (91.9%) out of 160 vivax malaria, 12 (92.3%) out of 13 falciparum malaria, and all 9 vivax/falciparum mixed malaria reacted with at least one antigen, while no reactions occurred with 20 normal uninfected sera. In the case of vivax malaria, CSP-1 reacted with 128 (80.0%) sera, MSP-1 with 102 (63.8%), AMA-1 with 128 (80.0%), SERA with 115 (71.9%), and EXP-1 with 89 (55.6%), respectively. We obtained higher detection rates when using S antigens (91.9%) rather than using each antigen solely (55.6 - 80%), a combination of 2 (76.3 - 87.5%), 3 (85.6 - 90.6%), or 4 antigens (89.4 - 91.3%). This method can be applied to serological diagnosis, mass screening in endemic regions, or safety test in transfusion of prevalent vivax malaria.

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Study on the pathogenesis of canine herpesvirus infection II. Immunohistochemical observation (Canine herpesvirus 감염증의 병리발생에 관한 연구 II. 면역조직화학적 관찰)

  • Seo, Il-bok;Lim, Chang-hyeong
    • Korean Journal of Veterinary Research
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    • v.34 no.3
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    • pp.583-591
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    • 1994
  • This study was carried out to investigate the pathogenesis of canine herpesvirus(CHV) infection in dogs. The 17 puppies, one day old, delivered from CHV seronegative 3 dams were divided into two groups. The 13 puppies were inoculated intranasally with 1ml of CHV-KK inoculum($5{\times}10^{5.6}TCID_{50}/ml$) and 4 puppies were served as control. And then the puppies were sacrificed at 2, 4, 6 and 7 days after the treatment, and sampled nasal mucosa, trigeminal nerve, trigeminal ganglion, bone marrow, eye, brain and other major organs for the immunohistochemical examination. Distribution of CHV antigens was limited in cytoplasms and nuclei of necrotic nasal epthelia at 2 days after infection. At 4 days after infection, CHV antigens were detected in vascular walls and peripheral nerves of nasal lamina propria, reticuloendothelial cells of spleen, interstitium of kidney, leptomeningeal vascular walls and alveolar walls, At 6 and 7 days after infection, CHV antigens were detected in all of the necrotic area. CHV antigens were also detected in vascular endothelial cells of various organs and in blood leukocytes from 4 days after infection. Among the six puppies in which necrotic lesions of central nervous system were observed, CHV antigens were detected in trigeminal ganglion, trigeminal nerve and ventroposteriomedial nucleus of four puppies and in spinal trigeminal nucleus of three puppies. These results indicate that the generalized focal necrosis of all organs including brain and eyes in canine herpesvirus infection were resulted from generalized vasculitis with leukocyte-associated viremia, and also the hemonecrotizing meningoencephalitis was resulted from spreading of CHV via blood and nerve trunk.

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Cancer/Testis OIP5 and TAF7L Genes are Up-Regulated in Breast Cancer

  • Mobasheri, Maryam Beigom;Shirkoohi, Reza;Modarressi, Mohammad Hossein
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.11
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    • pp.4623-4628
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    • 2015
  • Breast cancer still remains as the most frequent cancer with second mortality rate in women worldwide. There are no validated biomarkers for detection of the disease in early stages with effective power in diagnosis and therapeutic approaches. Cancer/testis antigens are recently promising tumor antigens and suitable candidates for targeted therapies and generating cancer vaccines. We conducted the present study to analyze transcript changes of two cancer/testis antigens, OIP5 and TAF7L, in breast tumors and cell lines in comparison with normal breast tissues by quantitative real time RT-PCR for the first time. Significant over-expression of OIP5 was observed in breast tumors and three out of six cell lines including MDA-MB-468, T47D and SKBR3. Not significant expression of TAF7L was evident in breast tumors but significant increase was noted in three out of six cell lines including MDA-MB-231, BT474 and T47D. OIP5 has ssignificant role in chromatin organization and cell cycle control during cell cycle exit and normal chromosome segregation during mitosis and TAF7L is a component of the transcription factor IID, which is involved in transcription initiation of most protein coding genes. TAF7Lis located at X chromosome and belongs to the CT-X gene family of cancer/testis antigens which contains about 50% of CT antigens, including those which have been used in cancer immunotherapy.

Protective efficacy of vaccination with Neospora caninum multiple recombinant antigens against experimental Neospora caninum infection

  • CHO Jung-Hwa;CHUNG Woo-Suk;SONG Kyoung-Ju;NA Byoung-Kuk;KANG Seung-Won;SONG Chul-Yong;KIM Tong-Soo
    • Parasites, Hosts and Diseases
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    • v.43 no.1 s.133
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    • pp.19-25
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    • 2005
  • Protective efficacy of vaccination with Neospora caninum multiple recombinant antigens against N. caninum infection was evaluated in vitro and in vivo. Two major immunodominant surface antigens (NcSAG1 and NcSRS2) and two dense granule proteins (NcDG1 and NcDG2) of N. caninum tachyzoites were expressed in E. coli, respectively. An in vitro neutralization assay using polyclonal antisera raised against each recombinant antigen showed inhibitory effects on the invasion of N. caninum tachyzoites into host cells. Separate groups of gerbils were immunized with the purified recombinant proteins singly or in combinations and animals were then challenged with N. caninum. Following these experimental challenges, the protective efficacy of each vaccination was determined by assessing animal survival rate. All experimental groups showed protective effects of different degrees against experimental infection. The highest protection efficacy was observed for combined vaccination with NcSRS2 and NcDG1. Our results indicate that combined vaccination with the N. caninum recombinant antigens, NcSRS2 and NcDG1, induces the highest protective effect against N. caninum infection in vitro and in vivo.

Immunogenicity of a DNA and Recombinant Protein Vaccine Combining LipL32 and Loa22 for Leptospirosis Using Chitosan as a Delivery System

  • Umthong, Supawadee;Buaklin, Arun;Jacquet, Alain;Sangjun, Noppadol;Kerdkaew, Ruthairat;Patarakul, Kanitha;Palaga, Tanapat
    • Journal of Microbiology and Biotechnology
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    • v.25 no.4
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    • pp.526-536
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    • 2015
  • Leptospirosis is a worldwide zoonotic disease caused by pathogenic Leptospira, a genus of which more than 250 serovars have been identified. Commercial bacterin vaccines are limited in that they lack both cross-protection against heterologous serovars and long-term protection. This study investigated in mice the immunogenicity of an anti-leptospirosis vaccine, using the outer membrane proteins LipL32 and Loa22 as antigens. The immunogenicity of this vaccine formulation was compared with those induced by vaccines based on LipL32 or Loa22 alone. A DNA-encapsulated chitosan nanoparticle was used for in vivo DNA delivery. Using a unique DNA plasmid expressing both lipL32 and loa22 for vaccination, higher antibody responses were induced than when combining plasmids harboring each gene separately. Therefore, this formulation was used to test the immunogenicity when administered by a heterologous prime (DNA)-boost (protein) immunization regimen. The specific antibody responses against LipL32 (total IgG and IgG1) and Loa22 (IgG1) were higher in mice receiving two antigens in combination than in those vaccinated with a single antigen alone. Although no significant difference in splenic CD4+ T cell proliferation was observed among all groups of vaccinated mice, splenocytes from mice vaccinated with two antigens exhibited higher interferon-γ and IL-2 production than when using single antigens alone upon in vitro restimulation. Taken together, the immunogenicity induced by LipL32 and Loa22 antigens in a heterologous primeboost immunization regimen using chitosan as a DNA delivery system induces higher immune response, and may be useful for developing a better vaccine for leptospirosis.