• Molecules and Cells
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• v.45 no.12
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• pp.896-910
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• 2022

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly transmissible and potentially fatal virus. So far, most comprehensive analyses encompassing clinical and transcriptional manifestation have concentrated on the lungs. Here, we confirmed evident signs of viral infection in the lungs and spleen of SARS-CoV-2-infected K18-hACE2 mice, which replicate the phenotype and infection symptoms in hospitalized humans. Seven days post viral detection in organs, infected mice showed decreased vital signs, leading to death. Bronchopneumonia due to infiltration of leukocytes in the lungs and reduction in the spleen lymphocyte region were observed. Transcriptome profiling implicated the meticulous regulation of distress and recovery from cytokine-mediated immunity by distinct immune cell types in a time-dependent manner. In lungs, the chemokine-driven response to viral invasion was highly elevated at 2 days post infection (dpi). In late infection, diseased lungs, post the innate immune process, showed recovery signs. The spleen established an even more immediate line of defense than the lungs, and the cytokine expression profile dropped at 7 dpi. At 5 dpi, spleen samples diverged into two distinct groups with different transcriptome profile and pathophysiology. Inhibition of consecutive host cell viral entry and massive immunoglobulin production and proteolysis inhibition seemed that one group endeavored to survive, while the other group struggled with developmental regeneration against consistent viral intrusion through the replication cycle. Our results may contribute to improved understanding of the longitudinal response to viral infection and development of potential therapeutics for hospitalized patients affected by SARS-CoV-2.

• Pediatric Infection and Vaccine
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• v.6 no.2
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• pp.253-260
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• 1999

Purpose : The purpose of this study is to make and use monoclonal Ab which reacts with CMV major immediate early(${\alpha}$) protein(p72). Methods : Normal human fibroblast(Foreskin derived) was cultured in Eagle's minimal essential medium(MEM) containing 10% cowfetus serum and mouse chondroblast was cultured in P3X63 Ag8.653(ATCC. Maryland USA) to maintain $5{\times}10^5/ml$ cell counts. CMV(KJHJ90) from congenital CMV infected infant's urine was multiplied and used for Ab making. CMV Ag was injected 4 times, 1 week interval into the peritoneal space of 6~8 weeks old mice. And then lymphocyles and fibroblasts taken from spleen were obtained and conjugated. After the conjugated cell cultured, we chose the cell that had high Ab titer using indirect immunofluerescent method. Results : Among the 28 monoclonal antibodies obtained LPC12 and LPC23 reacted highly with nucleus of AD169 infected cell. Purified AD169 after SDS-PAGE, molecular weight of Ag, which reacted with purified monoclonal Ab, was obtained using Western blotting. Monoclonal Ab of LPC12 and LPC23 clone reacted most highly with 72 kd Ag. Conclusion : LPC12 and LPC23 clonal Ab with AD 169(P63-27) is useful on early diagnosis of CMV infection.

• Korean Journal of Poultry Science
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• v.45 no.1
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• pp.17-28
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• 2018

Infectious bronchitis virus (IBV) is an economically important disease in the poultry industry worldwide. This disease commonly manifests respiratory signs, poor egg quality, and decline in egg production. Since IBV is a RNA virus, the emergence of new variant strains is continuously reported and the immunization of susceptible chickens with only one antigenic type of the virus has been shown to induce partial or no protection against other unrelated types. Therefore, it is difficult to diagnose IBV due to variants serotypes. In this study, we collected serum from various ages of Broiler GP (Grandparent) to Layer CC (Commercial chick) and performed detectability comparison test between domestic company and multinational company manufacturing ELISA kit. Results of this experiment suggest that domestic company manufacturing ELISA kit is more sensitive to infectious bronchitis antibody than that of the multinational company. Our findings also suggest that antibody's change trends after infectious bronchitis vaccination. Thus, the use of appropriate kit for domestic situations is important.

• Journal of Plant Biotechnology
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• v.31 no.4
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• pp.285-293
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• 2004

FMDV is a viral pathogen that caused foot-and-mouth disease in animals. VP1 is a major capsid protein of FMDV. It is known as one of best materials for the FMDV diagnosis and for the development of protein vaccine. In this study, 633 bp of VP1 gene was modified for the expression of VP1 in plant, based on the VP1 DNA sequence from FMDV taiwan O type and from FMDV isolated vietnam. The. deduced DNA fragment was artificially synthesized using the multiple fragment extension with long-nucleotides. A new plant transgenic vector system, pCAMBIA139011 was constructed on the basis of pBI12l and pCAMBIA1390. Using this vector system and GFP gene or modified VP1 gene, each target gene was introduced into Nicotiana tabacum. The insertion of whole target gene was successfully confirmed in each transgenic plant named GFP-A7 and VP1-4, respectively. The expression level of each gene was estimated by RT-PCR and Real-Time PCR using VP1, GFP specific primers.

• Parasites, Hosts and Diseases
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• v.55 no.2
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• pp.149-158
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• 2017

Variant surface antigens (VSAs) encoded by pir families are considered to be the key proteins used by many Plasmodium spp. to escape the host immune system by antigenic variation. This attribute of VSAs is a critical issue in the development of a novel vaccine. In this regard, a population genetic study of vir genes from Plasmodium vivax was performed in the Republic of Korea (ROK). Eighty-five venous blood samples and 4 of the vir genes, namely vir 27, vir 21, vir 12, and vir 4, were selected for study. The number of segregating sites (S), number of haplotypes (H), haplotype diversity (Hd), DNA diversity (${\pi}$ and ${\Theta}_w$), and Tajima's D test value were conducted. Phylogenetic trees of each gene were constructed. The vir 21 (S=143, H=22, Hd=0.827) was the most genetically diverse gene, and the vir 4 (S=6, H=4, Hd=0.556) was the opposite one. Tajima's D values for vir 27 (1.08530, P>0.1), vir 12 (2.89007, P<0.01), and vir 21 (0.40782, P>0.1) were positive, and that of vir 4 (-1.32162, P>0.1) was negative. All phylogenetic trees showed 2 clades with no particular branching according to the geographical differences and cluster. This study is the first survey on the vir genes in ROK, providing information on the genetic level. The sample sequences from vir 4 showed a clear difference to the Sal-1 reference gene sequence, whereas they were very similar to those from Indian isolates.

• Korean Journal of Microbiology
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• v.39 no.2
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• pp.89-94
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• 2003

Respiratory viruses were isolated from patients with acute respiratory infections in Busan during 2000-2001 and characterized for their antigenic properties. In 2000, 39 out of 43 isolated viruses were identified as influenza viruses and the others were adenoviruses. Among the isolated influenza viruses,23 were type A influenza viruses and 16 were type B influenza viruses. As a result of antigenic characterization, the influenza viruses were determined to A/Sydney/05/97(H3N2)-like, A/Beijing/262/95(H1N1)-like, and B/Harbin07/94-like viruses and serotypes of the isolated adenoviruses were type 1, 2, and 5. In 2001, 56 viruses were isolated and all of the viruses were identified as influenza viruses. They were A/panama/253/99(H3N2)-like and A/Newcaledonia/2007/99(H1Nl)-like viruses when determined by their antigenic properties. The sex distribution of the patients is as follows, 14 males (32.56%),23 females (67.44％) in 2000, and 23 males (41.07%), 33 females (58.93%) in 2001. Occurrence rate was found to be higher in female patients in both years. Age distribution of patients, in 2000, 48.84％ of infection occurred in 0 to 1 year old while in 2002, 33.93% occurred among 11-20 year olds. In 2000, occurrence rate was found to be high in January and again in April and various types of viruses were isolated. These results may be useful for vaccine development and establishment of reliable epidemic data.

• Pediatric Infection and Vaccine
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• v.7 no.1
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• pp.152-158
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• 2000

Purpose : Kawasaki disease is an acute illness of unknown cause that affects infants and children. Sometimes, cervical lymphadenitis that seems to be caused by bacterial or viral agents at admission would be changed to Kawasaki disease within several days of hospitalization. Lymphadenopathy is one of major presenting manifestation of both cervical lymphadenitis and Kawasaki disease. We compared Kawasaki disease with cervical lymphadenopathy as the initial manifestation with cervical lymphadenitis without other typical manifestations of Kawasaki disease. Methods : We describe and compare the clinical characteristics of 15 patients with Kawasaki disease which were misdiagnosed as cervical lymphadenitis and 125 patients with cervical lymphadenitis, retrospectively. Results : The clinical characteristics of Kawasaki disease with cervical lymphadenopathy as an initial presenting manifestation were older in age and had more frequent abnormalities in markers of systemic inflammation than kawasaki disease without cervical lymphadenopathy. They would have greater abnormalities in markers of inflammation than cervical lymphadenitis alone. They had higher mean WBC, cESR, CRP which were compared with the patients with cervical lymphadenitis. Usually, patients with cervical lymphadenitis responded to antibiotics promptly, but they did not tend to be responded to antibiotics within 48~72hrs. Conclusion : Patients with Kawasaki disease may initially present with findings that strongly suggest bacterial lymphadenitis with or without other manifestations of Kawasaki disease. If they did not response initial antibiotics administration, they should be closed observed for the possible development of the another signs of Kawasaki disease. Earlier diagnosis of Kawasaki disease can avoid the need for unnecessary procedures and antibiotics and lead to more proper therapy directed at Kawasaki disease.

• Parasites, Hosts and Diseases
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• v.54 no.4
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• pp.385-391
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• 2016

The discovery and understanding of antigenic proteins are essential for development of a vaccine against malaria. In Plasmodium falciparum, Pf92 have been characterized as a merozoite surface protein, and this protein is expressed at the late schizont stage, but no study of Pv92, the orthologue of Pf92 in P. vivax, has been reported. Thus, the protein structure of Pv92 was analyzed, and the gene sequence was aligned with that of other Plasmodium spp. using bioinformatics tools. The recombinant Pv92 protein was expressed and purified using bacterial expression system and used for immunization of mice to gain the polyclonal antibody and for evaluation of antigenicity by protein array. Also, the antibody against Pv92 was used for subcellular analysis by immunofluorescence assay. The Pv92 protein has a signal peptide and a sexual stage s48/45 domain, and the cysteine residues at the N-terminal of Pv92 were completely conserved. The N-terminal of Pv92 was successfully expressed as soluble form using a bacterial expression system. The antibody raised against Pv92 recognized the parasites and completely merged with PvMSP1-19, indicating that Pv92 was localized on the merozoite surface. Evaluation of the human humoral immune response to Pv92 indicated moderate antigenicity, with 65% sensitivity and 95% specificity by protein array. Taken together, the merozoite surface localization and antigenicity of Pv92 implicate that it might be involved in attachment and invasion of a merozoite to a new host cell or immune evasion during invasion process.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.9
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• pp.3595-3604
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• 2015

Hepatocellular carcinoma (HCC) has been one of the most fatal malignant tumors worldwide and its associated morbidity and mortality remain of significant concern. Based on in-depth reviews of serological diagnosis of HCC, in addition to AFP, there are other biomarkers: Lens culinaris agglutinin-reactive AFP (AFP-L3), descarboxyprothrombin (DCP), tyrosine kinase with Ig and eprdermal growth factor (EGF) homology domains 2 (TIE2)-espressing monocytes (TEMs), glypican-3 (GPC3), Golgi protein 73 (GP73), interleukin-6 (IL-6), and squamous cell carcinoma antigen (SCCA) have been proposed as biomarkers for the early detection of HCC. The diagnosis of HCC is primarily based on noninvasive standard imaging methods, such as ultrasound (US), dynamic multiphasic multidetector-row CT (MDCT) and magnetic resonance imaging (MRI). Some experts advocate gadolinium diethyl-enetriamine pentaacetic acid (Gd-EOB-DTPA) MRI and contrast-enhanced US as the promising imaging madalities of choice. With regard to recent advancements in tissue markers, many cuting-edge technologies using genome-wide DNA microarrays, qRT-PCR, and proteomic and inmunostaining studies have been implemented in an attempt to identify markers for early diagnosis of HCC. Only less than half of HCC patients at initial diagnosis are at an early stage treatable with curative options: local ablation, surgical resection, or liver transplant. Transarterial chemoembolization (TACE) is considered the standard of care with palliation for intermediate stage HCC. Recent innovative procedures using drug-eluting-beads and radioembolization using Yttrium-90 may exhibit beneficial effects in HCC treatment. During the past few years, several molecular targeted agents have been evaluated in clinical trials in advanced HCC. Sorafenib is currently the only approved systemic treatment for HCC. It has been approved for the therapy of asymptomatic HCC patients with well-preserved liver function who are not candidates for potentially curative treatments, such as surgical resection or liver transplantation. In the USA, Europe and particularly Japan, hepatitis C virus (HCV) related HCC accounts for most liver cancer, as compared with Asia-Pacific regions, where hepatitis B virus (HBV) may play a more important role in HCC development. HBV vaccination, while a vaccine is not yet available against HCV, has been recognized as a best primary prevention method for HBV-related HCC, although in patients already infected with HBV or HCV, secondary prevention with antiviral therapy is still a reasonable strategy. In addition to HBV and HCV, attention should be paid to other relevant HCC risk factors, including nonalcoholic fatty liver disease due to obesity and diabetes, heavy alcohol consumption, and prolonged aflatoxin exposure. Interestingly, coffee and vitamin K2 have been proven to provide protective effects against HCC. Regarding tertiary prevention of HCC recurrence after surgical resection, addition of antiviral treatment has proven to be a rational strategy.

• Pediatric Infection and Vaccine
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• v.20 no.1
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• pp.28-35
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• 2013

Purpose : The aim of this study was to determine the frequency, epidemiology and the clinical manifestation of human metapneumovirus (hMPV) infection in Korean children. Methods : From February 2010 to January 2012, we collected nasopharyngeal aspiration from 1,554 children who were hospitalized for acute lower respiratory tract infections at the Department of Pediatrics, Severance Children's Hospital. hMPV was detected by performing reverse transcriptase-polymerase chain reaction (RT-PCR). The medical records of the patients with positive results were retrospectively reviewed. Results : We detected hMPV in 99 of the 1,554 hospitalized children. The mean age of the hMPV infected children was 25 months, and 87% of the illnesses occurred between April and June. The most common diagnoses were pneumonia (73%) and bronchiolitis (16%). The clinical manifestations included cough, fever, respiratory distress, hoarseness, tachypnea, and wheezing. Coinfection with other respiratory viruses was found in 43 children (43%). Conclusion : hMPV is one of the major virus causing acute respiratory tract infection in the age between 13 months and 48 months old with peaks during April to June. Reports of hMPV in Korea has been increasing but additional studies are required to define the epidemiology and the extent of disease caused by hMPV to determine future development of this illness in Korean children.