• Journal of Microbiology and Biotechnology
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• v.23 no.8
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• pp.1159-1166
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• 2013

Orientia tsutsugamushi, a gram-negative bacterium, causes severe acute febrile illness in humans. Despite this danger, the route of infection, infectivity, and protective mechanisms of the host's immune response to O. tsutsugamushi are unclear. Dendritic cells (DCs) are one of the most important cell types in bridging the innate and adaptive immune responses. In this study, we observed that O. tsutsugamushi infects and replicates in monocyte-derived DCs (MODCs). During infection and replication, the expressions of the cytokines IL-12 and TNF-${\alpha}$, as well as the co-stimulatory molecules CD80, CD83, CD86, and CD40, were increased in MODCs. When O. tsutsugamushi-treated MODCs were co-cultured with autologous $CD4^+$ T cells, they enhanced production of IFN-${\gamma}$, a major Th1 cytokine. Collectively, our results show that O. tsutsugamushi can replicate in MODCs and can simultaneously induce MODC maturation and increase proinflammatory cytokine levels in MODCs that subsequently activate $CD4^+$ T cells.

• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.101-108
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• 2020

Infections by herpes simplex viruses have an immense impact on humans, ranging from self-limiting, benign illness to serious, life-threatening diseases. While nucleoside analog drugs are available, resistance has been increasing and currently no vaccine exists. Ginsenosides derived from Panax ginseng have been documented to inhibit several viruses and bolster immune defenses. This study evaluated 12 of the most relevant ginsenosides from P. ginseng for toxicities and inhibition of herpes simplex viruses types 1 and 2 in Vero cells. The effects of test compounds and virus infection were determined using a PrestoBlue cell viability assay. Time course studies were also conducted to better understand at what points the virus life cycle was affected. Non-toxic concentrations of the ginsenosides were determined and ranged from 12.5 μM to greater than 100 μM. Ginsenoside 20(S)-Rg3 demonstrated the greatest inhibitory effect and was active against both HSV-1 and HSV-2 with an IC₅₀ of approximately 35 μM. The most dramatic inhibition-over 100% compared to controls-occurred when the virus was exposed to 20(S)-Rg3 for 4 h prior to being added to cells. 20(S)-Rg3 holds promise as a potential chemotherapeutic agent against herpes simplex viruses and, when used together with valacyclovir, may prevent increased resistance to drugs.

• Korean Journal of Veterinary Research
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• v.41 no.1
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• pp.59-65
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• 2001

To clarify for the presence of infectious bronchitis virus (IBV) in Korea before 1986, in which the virus was first isolated, materials collected from chicken diagnostic consignments between 1980 and 1985 and propagated in chicken embryos or cell cultures were screened by the reverse transcriptase polymerase chain reaction (RT-PCR) targeted to the nucleocapsid gene of the virus. Among 11 samples examined, one sample (IBV-SNU80108) submitted in 1980 showed specific PCR product (281 bp). When the amplified product was sequenced, together with IBV vaccine virus H120 strain, and compared with the data for ten other IBV strains derived from the GeneBank, identities between IBV-SNU80108 and other strains in nucleotide and amino acid sequences ranged 96.3% to 63.7% and 96.4% to 69%, respectively. IBV-SNU80108 was distinct from H120 strain by showing 91.9% and 92.9% identities in the respective sequences. This data suggested that IBV genetically distinctive from other foreign IBV strains might be present before 1986 in Korea.

• Journal of Life Science
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• v.33 no.1
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• pp.64-72
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• 2023

Despite several advances in identification of cardiac transcription factors, there are still needs to find new bioactive molecules that promote cardiomyogenesis from stem cells to highly efficient myocardial differentiation. We analyzed Illumina expression microarray data of mouse embryonic stem cells (mESCs)-derived cardiomyocytes. 276 genes were upregulated (≥ 4fold) in mESCs-derived cardiomyocytes compared undifferentiated ESCs. Secreted phosphoprotein 2 (Spp2) is one of candidates and is known to inhibit bone morphogenetic protein 2 (BMP2) signal transduction as a pseudoreceptor for BMP2. However, its function in cardiomyogenesis is unknown. We confirmed that Spp2 expression increased during the differentiation into functional cardiomyocytes using mESCs, TC-1/Kh2 and E14. Interestingly, Spp2 secretion transiently increased 3 days after formation of embryoid bodies (EBs), indicating that the extracellular secretion of Spp2 is involved in the differentiation of ESCs into cardiomyocytes. To characterize Spp2, we performed experiments using the C2C12 mouse myoblast cell line, which has the property of shifting the differentiation pathway from myoblastic to osteoblastic by treatment with BMP2. Similar to the differentiation of ESCs, transcription of Spp2 increased as C2C12 myoblasts differentiated into myotubes. In particular, Spp2 secretion increased dramatically in the early stage of differentiation. Furthermore, treatment with Spp2-Flag recombinant protein promoted the differentiation of C2C12 myoblasts into myotubes. Taken together, we suggest a novel bioactive protein Spp2 that differentiates ESCs into cardiomyocytes. This may be useful for understanding the molecular pathways of cardiomyogenesis and for experimental or clinical promotion of stem cell therapy for ischemic heart diseases.

• IMMUNE NETWORK
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• v.4 no.1
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• pp.31-37
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• 2004

Background: 1-8D gene is a member of human 1-8 interferon inducible gene family and is shown to be overexpressed in fresh colon cancer tissues. Three peptides 1-6, 3-5 and 3-7 derived from 1-8D gene were shown to have immunogenicity against colon cancer. Methods: To study tumor immunotherapy of these peptides we established an adoptive transfer model. $D^{b-/-}{\times}{\beta}2$ microglobulin (${\beta}2m$) null mice transgenic for a chimeric HLA-A2.1/$D^b-{\beta}2m$ single chain (HHD mice) were immunized with irradiated peptide-loaded RMA-S/HHD/B7.1 transfectants. Spleens were removed after last immunization, and splenocytes were re-stimulated in vitro. Lymphocytes from vaccinated HHD mice were transferred together with IL-2 to the tumor bearing nude mice that were challenged S.C. with the HCT/HHD/B7 colon carcinoma cell line that was found to grow in these mice. Results: Peptide 3-5 was found to be highly effective in CTL activity. Adoptively transferred anti-peptide 3-5 cytolytic T lymphocytes caused significant retardation in tumor growth. Conclusion: This study shows that peptide 3-5 can be the most effective candidate for the vaccine of adoptive immunotherapy against colon cancer.

• IMMUNE NETWORK
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• v.14 no.1
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• pp.21-29
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• 2014

Follicular helper T ($T_{FH}$) cells are recently highlighted as their crucial role for humoral immunity to infection as well as their abnormal control to induce autoimmune disease. During an infection, na$\ddot{i}$ve T cells are differentiating into $T_{FH}$ cells which mediate memory B cells and long-lived plasma cells in germinal center (GC). $T_{FH}$ cells are characterized by their expression of master regulator, Bcl-6, and chemokine receptor, CXCR5, which are essential for the migration of T cells into the B cell follicle. Within the follicle, crosstalk occurs between B cells and $T_{FH}$ cells, leading to class switch recombination and affinity maturation. Various signaling molecules, including cytokines, surface molecules, and transcription factors are involved in $T_{FH}$ cell differentiation. IL-6 and IL-21 cytokine-mediated STAT signaling pathways, including STAT1 and STAT3, are crucial for inducing Bcl-6 expression and $T_{FH}$ cell differentiation. $T_{FH}$ cells express important surface molecules such as ICOS, PD-1, IL-21, BTLA, SAP and CD40L for mediating the interaction between T and B cells. Recently, two types of microRNA (miRNA) were found to be involved in the regulation of $T_{FH}$ cells. The miR-17-92 cluster induces Bcl-6 and $T_{FH}$ cell differentiation, whereas miR-10a negatively regulates Bcl-6 expression in T cells. In addition, follicular regulatory T ($T_{FR}$) cells are studied as thymus-derived $CXCR5^+PD-1^+Foxp3^+\;T_{reg}$ cells that play a significant role in limiting the GC response. Regulation of $T_{FH}$ cell differentiation and the GC reaction via miRNA and $T_{FR}$ cells could be important regulatory mechanisms for maintaining immune tolerance and preventing autoimmune diseases such as systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). Here, we review recent studies on the various factors that affect $T_{FH}$ cell differentiation, and the role of $T_{FH}$ cells in autoimmune diseases.

• 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.

• Journal of Life Science
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• v.25 no.11
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• pp.1197-1203
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• 2015

Salmonella Typhimurium (ST) JOL389 and χ3339 are strong virulent strains against mouse. ST χ8554 is derived by deletion of the asd gene from ST χ3339. Plasmid pMMP184 carrying a ghost cassette was transformed into ST χ8554, and ST χ8554 ghost cells were prepared and administrated via the oral route to BALB/c mice. Change in the amount of total IgG was not elicited to boosting of single ST χ8554 ghost cells, but the content was increased from 6 weeks after the 3^rd administration. However, when the ST JOL389 ghost cells is administered together with ST χ8554 ghost cells, the content of total IgG was increased in 2 weeks post primary administration. It was found that the content of total IgG of the group mixed with ST JOL389 ghost cells showed an increased value of 8 times or more at 10 weeks when compared with the group of ST χ8554 ghost cells. The content of IgG1, IgG2a, and sIgA in both groups increased from 4 weeks postprimary administration. As a challenge test of virulent ST χ3339, χ8554 (pMMP184) and χ8554 (pMMP184)/JOL389 ghost cell groups showed protection of 50% or more when compared to the control group. These results suggest that the preparation of combined ghost cells from a strong virulent ST increases immunity more than a single strain.

• Journal of Microbiology and Biotechnology
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• v.29 no.1
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• pp.160-170
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• 2019

The lactic acid bacteria species Lactobacillus plantarum (L. plantarum) has been used extensively for vaccine delivery. Considering to the critical role of dendritic cells in stimulating host immune response, in this study, we constructed a novel CD11c-targeting L. plantarum strain with surface-displayed variable fragments of anti-CD11c, single-chain antibody (scFv-CD11c). The newly designed L. plantarum strain, named 409-aCD11c, could adhere and invade more efficiently to bone marrow-derived DCs (BMDCs) in vitro due to the specific interaction between scFv-CD11c and CD11c located on the surface of BMDCs. After incubation with BMDCs, the 409-aCD11c strain harboring a eukaryotic vector pValac-GFP could lead to more efficient expression of GFP compared with wild-type strains shown by flow cytometry analysis, indicating the enhanced translocation of pValac-GFP from L. plantarum to BMDCs. Similar results were also observed in an in vivo study, which showed that oral administration resulted in efficient expression of GFP in both Peyer's patches (PP) and mesenteric lymph nodes (MLNs) within 7 days after the last administration. In addition, the CD11c-targeting strain significantly promoted the differentiation and maturation of DCs, the differentiation of $IL-4^+$ and $IL-17A^+$ T helper (Th) cells in MLNs, as well as production of $B220^+$ $IgA^+$ B cells in the PP. In conclusion, this study developed a novel DC-targeting L. plantarum strain which could increase the ability to deliver eukaryotic expression plasmid to host cells, indicating a promising approach for vaccine study.

• BMB Reports
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• v.44 no.11
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• pp.758-763
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• 2011

Immunostimulatory CpG-DNA targeting TLR9 is one of the most extensively evaluated vaccine adjuvants. Previously, we found that a particular form of natural phosphodiester bond CpG-DNA (PO-ODN) encapsulated in a phosphatidyl-${\beta}$-oleoyl-${\gamma}$-palmitoyl ethanolamine (DOPE) : cholesterol hemisuccinate (CHEMS) (1 : 1 ratio) complex (Lipoplex(O)) is a potent adjuvant. Complexes containing peptide and Lipoplex(O) are extremely useful for B cell epitope screening and antibody production without carriers. Here, we showed that IL-12 production was increased in bone marrow derived dendritic cells in a CpG sequence-dependent manner when PO-ODN was encapsulated in Lipoplex(O), DOTAP or lipofectamine. However, the effects of Lipoplex(O) surpassed those of PO-ODN encapsulated in DOTAP or lipofectamine and also other various forms of liposome-encapsulated CpG-DNA in terms of potency for protein antigen-specific IgG production and Th1- associated IgG2a production. Therefore, Lipoplex(O) may have a unique potent immunoadjuvant activity which can be useful for various applications involving protein antigens as well as peptides.