• Biomedical Science Letters
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• v.3 no.2
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• pp.89-94
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• 1997

The authors characterized the proteins of the crude antigen obtained from Clonorchis sinensis worm and excretory-secretory and billis from rabbits, experimentally infected for 3 months. Protein composition was observed after adding a cysteine proteinase inhibitor E-64 and a serine proteinase inhibitor PMSF, respectively. SDS-PAGE of the crude antigen from C. sinensis recovered from the infected rabbits, the crude antigen from the adult worm excretory-secretory, and the crude antigen from billis of the rabbits resolved 26, 27 and 19 profiles between 200-9 kDa, respectively. When E-64 supplemented 29, and 22 bands, respectively. More study should be carried out in the future on the immunological characteristics and the monoclonal antibody of the each antigen.

• IMMUNE NETWORK
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• v.7 no.3
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• pp.117-123
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• 2007

Background: This study was designed to investigate the role of the hepatocyte growth factor (HGF) with regards to differentiation of somatic stem cells originating from the human umbilical cord blood (UCB) into hepatic lineage cells in vitro culture system. Methods: Mononuclear cells from UCB were cultured with and without HGF based on the fibroblast growth factor (FGF)-1, FGF-2, and stem cell factor. The cultured cells were confirmed by immunofluorescent staining analysis with albumin (ALB), cytokeratin-19 (CK-19), and proliferating cell nuclear antigen (PCNA) MoAb. ALB and CK-18 mRNA were also evaluated by reverse transcription-polymerase chain reaction. In order to observe changes in proliferating capacity with respect to the cultured period, CFSE with affinity to proliferating cells were tagged and later underwent flow cytometry. Results: In the HGF-treated group, cultured cells had a large oval shaped appearance with adherent, but easily detachable characteristics. In the HGF-non treated group, these cells were spindle-shaped with strong adherent characteristics. Expressions of ALB and CK-19 were evident in HGF-treated group compared to non-expression of those in to HGF-non treated group. Dual immunostaining analysis of the ALB producing cells showed presence of PCNA in their nuclei, and ALB and CK-18 mRNA were detected on the 21st day of cultured cells in the HGF-treated group. Conclusion: Our findings suggest that HGF has a pivotal role in differentiating somatic stem cells of human UCB into hepatic lineage cells in vitro.

• Tuberculosis and Respiratory Diseases
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• v.59 no.3
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• pp.272-278
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• 2005

Background : The immune responses mediated by CD8+T cells are known to be significant in controlling M. tuberculosis infections. In order to determine the role of cytotoxic CD8+T cells in the protective immune mechanism in latently infected subjects, this study examined whether or not the cytotoxic immune responses of CD8+T cells specific to the M. tuberculosis somatic antigens are induced in BCG vaccinated healthy subjects. Methods : Cytotoxicity and $IFN-{\gamma}$ elispot assays were used to investigate the activities of CD8+T cells specific for the $thyA_{30-38}$ peptide epitope in circulating peripheral blood mononuclear cells (PBMC) from BCG-vaccinated HLA-A*0201 and A*0206 subjects. Results : The results indicate the cytotoxic and $IFN-{\gamma}$ immune responses of CD8+T cells specific for $thyA_{30-38}$ were induced in BCG vaccinated healthy subjects. Conclusion : The cytotoxic and $IFN-{\gamma}$ responses by CD8+T cells specific for the M. tuberculosis somatic antigens are induced in BCG-vaccinated subjects, and appear to be involved in the protective immune mechanism in latently infected people against a M. tuberculosis infection.

• Parasites, Hosts and Diseases
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• v.21 no.2
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• pp.257-264
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• 1983

In order to obtain more specific antigenic preparation for the diagnosis of human paragonlmiasis, crude saline extract of whole worm (=PwWWE), secretory.excretory components (PwSEC) and secretion-excretion-free somatic extract (PwSM) of 12 week-old ParagoninBus westermani were filtrated through Sephadex G-200 gel column. The adult Paragonimus worms were obtained from expevimentally infected doge. A total of 11 antigenic solutions was Iyophilised or diluted to adjust protein content of 1mg/ml. To evaluate the antigenicity of crude antigens and fractions, micro-ELISA was done with the sera from P westermani in(ected cases, C. sinensis infected cases and non-infected control cases to detect Paragonimus specific IgG antibody. The results were as follows: 1. When the PwWWE was filtrated through Sephadex G-200 gel, it was separated into three fractions; PwWWE Fr. 1, PwWWE Fr. 2 and PwWWE Fr. 3. The percentage of protein content was 28.0%, 21.6% and 50.4% respectively. The PwSM was also. separated into three fractions; PwSM Fr. 1, PwSM Fr. 2, PwSM Fr. 3. and their percentage of protein content was 41.3%, 38.6% and 20.1%. However, the PwSEC showed different fractionation pattern; i.e. fraction 1 (=PwSEC Fr.1) and 3 (PwSEC Fr. 3) without fraction 2. The percentage of protein content was 14.0% in PwSEC Fr. 1 and 86.0% in PwSEC Fr. 3. 2. When the antigenicity of each Paragonimus crude antigen and fractionated antigen was evaluated for specific IgG aritibody by micro-ELISA in 10 human paragonimiasis sera, PwSEC Fr, 1 was the most potent antigen showing the mean absorbance 1.98. The PwWWE Fr. 1, PwSEC, PwWWE were next to that: their mean absorbance were 1.72, 1.38 and 0.83 respectively. The antigenicity of fractions 2 and 3 was much weaker in binding specific IgG antibody. 3. When the antigens were reacted in micro-ELISA with 10 human clonorchiasis sera, most antigens showed weak reactivity. Each fraction 1 of crude antigens reacted higher than other fractions or crude antigens; the mean absorbance was 0.17 in fraction 1, but in others the absorbances were about 0.06. 4. With non-infected control sera, the result of micro-RLISA revealed almost same pattern with those of the clonorchiasis sera. From the above results, it became apparent that PwWWE Fr. 1, especially PwSEC Fr. 1 was the most potent antigen reacted with Paragonisfaus specific IgG antibody.

• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.177-188
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• 2001

Mastitis is one of the most significant cause of economic loss to the dairy industry. Especially, Staphylococcus aureus is a major contagious mastitis-causing pathogen in dairy cattle. Because of its high transmission rate and resistance to antibiotic therapy, staphylococcal mastitis presents a constant threat to the dairy industry. Staphylococcal enterotoxin C(SEC) produced by S aureus has been known as one of superantigens which are able to stimulate a large proportion of T lymphocytes independently of their antigenic specificity. In this experiment, we have conducted preliminary studies with mice and lactating cows to evaluate the immunogenicity and safety of the experimental vaccine consists of SEC mutant antigen on controlling the bovine mastitis associated with S aureus infections. The average value of somatic cell counts in quarter milk, isolation rate of S aureus were consistently decreased in SEC-SER vaccinated groups, whereas antibody titers were highly increased in SEC-SER vaccinated groups. Peripheral blood were also collected from the lactating cows to determine the proportion of leukocyte subpopulation associated with humoral immunity(HI) and cell mediated immunity(CMI). Proportion of leukocyte subpopulation expressing $BoCD2^+$(total T lymphocyte), $BoCD4^+$(T helper cell), $BoCD8^+$(T cytotoxic/suppressor cell) and NonT/NonB lymphocyte which are involved in CMI in SEC-SER vaccinated groups were decreased for the initial stage after first vaccination and then increased from ten weeks after first vaccination maintaining elevated level till 14 weeks after vaccination. In contrast, proportion of monocyte, MHC class II and B lymphocyte which are associated with the production of primary immune response in SEC-SER vaccinated groups were increased for the initial period and then decreased from ten weeks after first vaccination. We present evidence that vaccination of SEC-SER mutant antigen in lactating cows induced a significant proliferation of bovine T lymphocytes. These results suggest that SEC-SER mutant antigen used in this experiment might be one of potential immunogen in developing innovative vaccine against bovine IMI associated with S aureus. Additional challenge trials should be carried out to evaluate substantial protection against S aureus under the commercial farm conditions.

• Korean Journal of Veterinary Research
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• v.6 no.1
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• pp.10-17
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• 1966

The antigenecity of somatic substances of S. pullorum standard strain and variant strain extracted byheat treatment, acid treatment and their modification, ammonium sulfate saturation (60 per cent), trypsin digestion was tested by indirest hemagglutination test and precipitation test and following results were optained. 1. Teatment at $100^{\circ}C$ for an hour of the bacteria could extract the antigen of S. pullorum standard strain and variant strain which was demonstrable by hemagglutination reaction with the human a group and chicken red blood cell. 2. Trypsin digestion was more enhanced its antigenecity in acid extracted antigen of S. pullorum variant strain compare with the S. pullorum standard strain. 3. The extracted antigenic substances of S. pullorum standard strain existed chiefly in the elicited fraction of precipitate at the treatment of ammonium sulfate saturation and after trypsin digestion, its antigenecity was demonstrated by hemagglutination. 4. At the treatment of ammonium sulfate treatment, did not occur the precipicate in acid extracted antigens of S. pullorum variant strain, however, the heal extracted antigen, positive reactions were obtained in both of the precipitate and supernatant fraction of the S. pullorum variant strain by hemagglutination reaction. After trypsin digestion, these fraction also exhibited positive reactions. 5. Precipitation test also tested dub could not detect in any soft of the antigens.

• Horticultural Science & Technology
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• v.35 no.1
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• pp.121-130
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• 2017

This study was carried out to investigate a method for producing cultured virus - free ovules for breeding high - quality Citrus cultivars. Ovules from the immature fruits of three citrus cultivars native to Jeju (Dongjeongkyool, Cheongkyool, and Jikak) and two cultivars of Citrus unshiu Marc. (Miyagawa wase and Haryejosaeng) that were thought to be infected with Citrus tristeza virus (CTV) were cultured on MS2 medium (Murashige - Skoog [MS] basal medium containing $500mg{\cdot}L^{-1}$ malt extract, $50g{\cdot}L^{-1}$ sucrose, $1.0 mg{\cdot}L^{-1}$ kinetin, and $8g{\cdot}L^{-1}$ agar). After four weeks of culture, 10, 21, 13, 5, and 7 somatic embryos and 2, 4, 2, 4, and 5 white callus cells (surrounding green somatic embryos) were obtained from Dongjeongkyool, Cheongkyool, Jikak, Miyagawa wase, and Haryejosaeng, respectively. After six weeks of culture, somatic embryos were obtained from cultured cells grown on MT basal medium supplemented with malt extract ($500mg{\cdot}L^{-1}$), lactose ($70g{\cdot}L^{-1}$), and agar ($16g{\cdot}L^{-1}$). Over 60% of the somatic embryos from citrus cultivars native to Jeju developed into normal plants on MS basal medium supplemented with malt extract ($500mg{\cdot}L^{-1}$), sucrose ($50g{\cdot}L^{-1}$), and agar ($8g{\cdot}L^{-1}$) after 10 weeks of culture. Normal plants were regenerated from two Citrus unshiu Marc. cultivars on MT basal medium supplemented with sorbitol (1.0 M), galactose (1.0 M), $GA_3$ ($1.0mg{\cdot}L^{-1}$), and Gelrite ($3g{\cdot}L^{-1}$). The absence of virus in plants generated from cultured ovules was confirmed by RT - PCR and antigen - antibody reactions. Therefore, virus - free Citrus cells can be obtained for breeding high - quality citrus cultivars using the biotechnological technique evaluated in this study.

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.263-268
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• 2005

Porcine epidemic diarrhea virus (PEDV) causes acute enteritis in pigs of all ages and is often fatal for neonates. In order to develop sweetpotato plants expressing PEDV antigen, we constructed the vector expressing spike gene of PEDV under the control of sweetpotato sporamin promoter or constitutive CaMV 35S promoter. The spike protein region of PEDV was synthesized by PCR and linked to each promoter, Transgenic sweetpotato [Ipomoea batatas (L.) Lam. cv. Yulmi] plants were developed from embryogenic calli following Agrobacterium tumefaciens-mediated transformation. The co-cultured embryogenic calli transferred to selective MS medium containing 1 mg/L 2,4-D, 100 mg/L kanamycin, and 400 mg/L claforan. These embryogenic calli were subcultured to the same selection medium at 3 weeks interval. Kanamycin-resistant calli transferred to hormone-free MS medium with kanamycin gave rise to somatic embryos and then converted into plantlets in the same medium. Southern blot analysis confirmed that the spike gene of PEDV was inserted into the genome of the sweetpotato plants. RT-PCR revealed that the spike gene of PEDV was highly expressed in transgenic sweetpotato plants.

• Molecules and Cells
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• v.40 no.9
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• pp.655-666
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• 2017

We constructed a large $na{\ddot{i}}ve$ human Fab library ($3{\times}10^{10}$ colonies) from the lymphocytes of 809 human donors, assessed available diversities of the heavy-chain variable (VH) and ${\kappa}$ light-chain variable (VK) domain repertoires, and validated the library by selecting Fabs against 10 therapeutically relevant antigens by phage display. We obtained a database of unique 7,373 VH and 41,804 VK sequences by 454 pyrosequencing, and analyzed the repertoires. The distribution of VH and VK subfamilies and germline genes in our antibody repertoires slightly differed from those in earlier published natural antibody libraries. The frequency of somatic hypermutations (SHMs) in heavy-chain complementarity determining region (HCDR)1 and HCDR2 are higher compared with the natural IgM repertoire. Analysis of position-specific SHMs in CDRs indicates that asparagine, threonine, arginine, aspartate and phenylalanine are the most frequent non-germline residues on the antibody-antigen interface and are converted mostly from the germline residues, which are highly represented in germline SHM hotspots. The amino acid composition and length-dependent changes in amino acid frequencies of HCDR3 are similar to those in previous reports, except that frequencies of aspartate and phenylalanine are a little higher in our repertoire. Taken together, the results show that this antibody library shares common features of natural antibody repertoires and also has unique features. The antibody library will be useful in the generation of human antibodies against diverse antigens, and the information about the diversity of natural antibody repertoires will be valuable in the future design of synthetic human antibody libraries with high functional diversity.

• Korean Journal of Poultry Science
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• v.15 no.3
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• pp.199-206
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• 1988

A total of 3 hybridoma clones producing monoclonal antibody (MCA) against infectious laryngotracheitis virus (ILTV) was established by somatic cell hybridization between mouse myeloma cells and spleen, cells from mice immunized with ILTV. The MCAs were screened by the indirect flourescent antibody (IFA) staining and the specific hybridomas were cloned by limiting dilution method. The MCAs produced by the 3 hybriomas were all classified as immunogloblin G and found to be reacting against common antigen(s) of high and low pathogenic ILTV examined. The titer of these antibodies in mouse ascitic fluid was from $10^5$ to $10^6$. Indirect fluorescent antibody test using these antibodies was found to be quite effective for the detection of ILTV from infected chickens being the most sensitive among the test methods adopted.