• Research in Plant Disease
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• v.25 no.3
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• pp.131-135
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• 2019

Cylindrocarpon destructans causes ginseng root rot and produces radicicol that has an antifungal effect. In this study, we developed a method to detect this fungus using enzyme-linked immunosorbent assay (ELISA). Secreted proteins of C. destructans were used as antigens to obtain C. destructans-specific IgG from mouse. Out of 318 monoclonal antibodies generated from mouse, two antibodies (Cd7-2-2 and Cd7-2-10) showed highest specificity and sensitivity. Indirect ELISA using both antigens successfully detected C. destructans in soils, but direct ELISA using IgG conjugated with horseradish peroxidase failed to detect antigens in soils. The indirect ELISA developed here can efficiently detect the fungus and help manage ginseng root rot disease in fields.

• Journal of fish pathology
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• v.37 no.1
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• pp.1-8
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• 2024

The advantages of replicon vectors of RNA viruses include a high ability to stimulate innate immunity and exponential amplification of target mRNA leading to high expression of foreign antigens. The present study aimed to construct a DNA-layered nervous necrosis virus (NNV) replicon vector system in which the capsid protein gene was replaced with a foreign antigen gene and to compare the efficiency of foreign antigen expression between the conventional DNA vaccine vector and the present replicon vector. We presented the first report of a nodavirus DNA replicon-based foreign antigen expression system. Instead of a two-vector system, we devised a one-vector system containing both an NNV RNA-dependent RNA polymerase cassette and a foreign antigen-expressing cassette. This single-vector approach circumvents the issue of low foreign protein expression associated with the low co-transfection efficiency of a two-vector system. Cells transfected with a vector harboring hammerhead ribozyme-fused RNA1 and RNA2 (with the capsid gene ORF replaced with VHSV glycoprotein ORF) exhibited significantly higher transcription of the VHSV glycoprotein gene compared to cells transfected with either a vector without hammerhead ribozyme or a conventional DNA vaccine vector expressing the VHSV glycoprotein. Furthermore, the transcription level of the VHSV glycoprotein in cells transfected with a vector harboring hammerhead ribozyme-fused RNA1 and RNA2 showed a significant increase over time. These results suggest that NNV genome-based DNA replicon vectors have the potential to induce stronger and longer expression of target antigens compared to conventional DNA vaccine vectors.

• BMB Reports
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• v.45 no.11
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• pp.595-603
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• 2012

Human Ly-6/uPAR molecules are a superfamily composed of two subfamilies; one is the membrane bound proteins with a GPI-anchor and the other are secreted proteins without the GPI-anchor. Ly-6/uPAR molecules have remarkable amino acid homology through a distinctive 8-10 cysteine-rich domain that is associated predominantly with O-linked glycans. These molecules are encoded by multiple tightly linked genes located on Chr. 8q23, and have a conserved genomic organization. Ly-6/uPAR molecules have an interesting expression pattern during hematopoiesis and on specific tumors indicating that Ly-6/uPAR molecules are associated with development of the immune system and carcinogenesis. Thus, Ly-6/uPAR molecules are useful antigens for diagnostic and therapeutic targets. This review summarizes our understanding of human Ly-6/uPAR molecules with regard to molecular structure as well as what is known about their function in normal and malignant tissues and suggest Ly-6/uPAR molecules as target antigens for cancer immunotherapy.

• Parasites, Hosts and Diseases
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• v.34 no.4
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• pp.239-246
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• 1996

The usefulness of malaria diagnosis by Plusmodium JaLcipawn-GDH (NADP+), obtained by affinity chromatography. is demonstrated in ELISA assays, testing IgG antibodies against GDH (NADP+) from patients with acute malaria, who have had two or more episodes of malaria. or from sera of hyperimmune patients. GDH (NADP+) thermal stability was demonstrated in a high heat resistance assay. The immunofluorescence assay demonstrated that anti-culture (P. falciporum) supernatant serum and anti-GDH (NADP+) of Proton app. recognized epitopes in Venezuelan isolates and Colombian and Brazilian malarial strains. The antigen is soluble, with high specificity is a potent imnlunogen and is thermoresistant. Key words: antigenic enzymes. glutamate dehydrogenase, malaria diagnosis, Plasmodium berghei, Plcswlodium ccthemelum, PlusmoniumJnlcipnmm, Plosmonium uiuox. soluble antigens.

• KSBB Journal
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• v.13 no.6
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• pp.649-655
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• 1998

Hantaviruses are rodent hosts-borne viruses belonging to the family Bunyaviridae, and are etiologic agents for two acute diseases, i.e., Haemorrhagic Fever with Renal Syndrome (HFRS) and Hantavirus Pulmonary Syndrome (HPS). There have been a lot of reports on prophylactic vaccines and diagnostics for the diseases, but most of viral antigens have been prepared by eukaryotic cell culture. Nucleocapsid proteins of Hantaviruses are known as the major viral antigens. Thereby, we prepared nucleocapsid genes of Hantaan virus and Seoul virus by RT-PCR and cloned into plasmid vectors, pET-3a and pKK223-3. Both genes were expressed in Escherichia coli with higher expression level of Seoul viral nucleocapsid protein compared to that of Hantaan in pET-3a. Hantaan viral gene was expressed much higher level in plasmid pET-3a that in pKK223-3. About 30% of expressed nucleocapsid protein was soluble and the rest was remained in insoluble fraction.

• Korean Journal of Veterinary Research
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• v.53 no.4
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• pp.199-205
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• 2013

Avian pathogenic Escherichia coli (APEC) is a causative agent for a number of extra intestinal diseases and account for significant losses to the poultry industry. Since protective immunity against APEC is largely directed to virulence antigens, we have individually expressed four different viulence antigens, papA, papG, IutA, and CS31A, using an attenuated Salmonella Typhimurium and a plasmid pBB244. Following oral immunization of mice with combination of two or four of these strains, serum IgG and mucosal IgA responses were elicited against each antigen represented in the mixture. The antigen-specific mucosal IgA responses were significantly higher in the group of mice immunized with the heat-labile Escherichia coli enterotoxin B subunit (LTB) strain than those in the group of mice immunized without the LTB strain. While, there was no significant difference between these two groups in antigen-specific serum IgG responses. The results showed that LTB could act as mucosal immune adjuvant. To assess the nature of immunity, the distribution of antigen-specific IgG isotypes was analyzed. All groups promoted Th1-type immunity as determined by the IgG2a/IgG1 ratio. Thus, our findings provided evidence that immunization with a combination of several vaccine strains is one of the strategies of developing effective vaccines against APEC.

• Korean Journal of Veterinary Service
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• v.43 no.4
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• pp.251-256
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• 2020

Two suckling piglets, 4 days and 10 days of age, showed lethargy and dyspnea after birth and mortality had been increased after incoming gilts from breeding farm. At necropsy, the lungs showed diffuse fail to collapse with rubbery consistency, edematous dilatation of interlobular septa, and lobular consolidation with purple red color. Heart was diffuse pale in color and had several irregular linear-shaped macules or patches. Histopathologically, diffuse interstitial pneumonia with the proliferation of type II pneumocytes was present in the lungs of 2 piglets. Alveolar lumens contained necrotic cellular debris derived from neutrophils and macrophages. Multifocal hemorrhage and necrotizing pneumonia with protozoan tachyzoites were observed in the lungs. Severe multifocal to confluent necrotic myocarditis, necrotic encephalitis, and necrotic adrenalitis with intralesional protozoan tachyzoites were observed in piglets. According to immunohistochemical analysis (IHC), Toxoplasma (T.) gondii tachyzoites antigens were confirmed in lung, heart, brain, and adrenal gland. And porcine reproductive and respiratory syndrome virus (PRRSV) antigens were also detected in the cytoplasm of macrophages in lungs using IHC. Based on the gross, histopathologic and immunohistochemical features, two suckling piglets were diagnosed as co-infection of T. gondii and PRRSV.

• Journal of Gastric Cancer
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• v.5 no.3 s.19
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• pp.180-185
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• 2005

Purpose: Among tumor-associated antigens, MAGE (melanoma antigen) was named as cancer/testis specific antigens because they are detected exclusively in the testis or cancer cells, including gastric carcinomas. Due to the elicitation of autoimmunitiy to tumors by these antigens either in vitro or in vivo and their tumor specificity, these antigens, thus, appear to be potential targets for tumor-specific immunotherapy. Materials and Methods: The fresh tumor tissue and normal gastric tissue samples were obtained from resected surgical specimens in 53 patients with gastric carcinomas. From the obtained cells, total cellular mRNA was extracted, and RT-PCR and nested PCR were run in 30 and 35 cycles respectively, with two different kinds of primers specially designed to detect six subtypes of MAGE DNA simultaneously. Results: In the 53 normal tissue, there was no expression of MAGE, but in the 53 cancer tissues, MAGE was expressed in 13 tissues (24.5%). Our data did not exhibit any correlation with the expression of the MAGE gene and clinicopathological factors. Conclusion: In our data, since 24.5% of gastric cancer tissues expressed MAGE, it should become possible to immunize a significant proportion of patients with advanced gastric carcinomas against the antigens encoded by these genes, provided that more antigenic peptides encoded by the genes of the MAGE family can be identified in the near future. (J Korean Gastric Cancer Assoc 2005;5:180-185)

• Korean Journal of Veterinary Research
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• v.38 no.1
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• pp.91-99
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• 1998

The immunogenetic analysis was performed to characterize the Korean native chickens (KNC) determined by monoclonal antibodies specific to chicken leukocyte differentiation antigens and flow cytometry. A total of 174 chickens including 58 KNC (black, brown and darkbrown colored), 77 foreign breed (Nagoya, White Reghorn, Rhode Island and Cornish) and 39 mixed breed (19 KNC with Nagoya and 20 KNC with Rhode Island) separately growing at Animal Science and Technology Institute were examined. The proportion of cells expressing MHC class II molecule (B-L in chicken) was significantly high in KNC. Proportion of CD4+ T helper cells was also higher in KNC and two mixed breed than that in foreign breed. However, proportion of CD8+ cells and TCR1 + (${\gamma}^{\delta}$ T cell receptor) cells was the lowest among the breed examined. Otherwise, those proportions were significantly high in White leghorn and two mixed breeds with two exclusive subpopulations. The two subpopulations were also typically shown in MHC class $II^+$ cells in KNC and one mixed breed, black-colored KNC with Nagoya. Although genotypic analysis was not pursued to characterize the immunogenetic properties of KNC, difference of phenotypic expression based on leukocyte differentiation molecules could be elucidated in KNC in this study.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.9
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• pp.1342-1346
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• 2006

Enterotoxigenic Escherichia coli is a major cause of diarrhea in neonatal and post-weaning piglets. To determine the most common fimbrial antigens of ETEC in piglets with diarrhea, two investigations were carried out on intensive pig farms in Hubei province, central China. In 2002-2003, 227 fecal samples from neonatal and post-weaning piglets with diarrhea were tested for the presence of the fimbrial antigen K88 and K99 of ETEC by the polymerase chain reaction (PCR). Twenty-three (10.1%) of 227 fecal samples were found to contain fimbrial antigen K88, which was identified as K88ac variant; and 13 (5.7%) samples containing K99. In 2004, another 179 fecal samples from diarrheic piglets, 1 day to 6 weeks of age, were tested for prevalence of fimbrial antigen K88, K99 and 987P. Forty-seven (26.3%) of the 179 samples carried at least one of the ETEC fimbrial antigens. K88 antigen was detected in 20.1%. In the 36 samples known to carry fimbrial antigen K88, 32 (88.9%) contained K88ad; and 4 (11.1%) contained K88ac; none of them carried K88ab. Fimbrial antigens K99 and 987P were detected in 1.1% and 6.1%, respectively. Our data indicate that K88 is the most common fimbrial antigen of ETEC associated with diarrhea in piglets in Central China.