• Title/Summary/Keyword: host cells

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Infection and Pathogenesis Mechanisms of Marek's Disease Virus (마렉병 바이러스 감염과 병원성 발현 기전)

  • Jang, H.K.;Park, Y.M.;Cha, S.Y.;Park, J.B.
    • Korean Journal of Poultry Science
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    • v.35 no.1
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    • pp.39-55
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    • 2008
  • Like the other herpesviruses, the virion of MDV consists of an envelope, which surrounds an amorphous tegument. Within the tegument, and icosahedral capsid encloses a linear double-stranded DNA core. Although the genome structure of MDV indicates that it is an ${\alpha}-herpesvirus$ like herpes simplex and varicella-zoster viruses, biological properties indicate MDV is more akin to the ${\gamma}-herpesvirus$ group, which includes Epstein-Barr and Kaposi's sarcoma herpesviruses. These herpesviruses replicate lytically in lymphocytes, epithelial and fibroblastic cells, and persist in lymphoblastoid cells. MDV has a complex life cycle and uses two means of replication, productive and non-productive, to exist and propagate. The method of reproduction changes according to a defined pattern depending on changes in virus-cell interactions at different stages of the disease, and in different tissues. Productive (lytic) interactions involve active invasion and take-over of the host cell, resulting in the production of infectious progeny virions. However, some herpesviruses, including MDV, can also establish a non-productive (abortive) infection in certain cell types, resulting in production of cell-associated progeny virus. Non-productive interactions represent persistent infection, in which the viral genome is present but gene expression is limited, there is no structural or regulatory gene translation, no replication, no release of progeny virions and no cell death. Reactivation of the virus is rare, and usually the infectious virus can be re-isolated only after cultivation in vitro. MDV establishes latency in lymphoid cells, some of which are subsequently transformed. In this review article, recent knowledges of the pathogenesis mechanisms followed by MDV infection to sensitive cells and chickens are discussed precisely.

Macrophage-secreted Exosomes Delivering miRNA-21 Inhibitor can Regulate BGC-823 Cell Proliferation

  • Wang, Jian-Jun;Wang, Ze-You;Chen, Rui;Xiong, Jing;Yao, Yong-Liang;Wu, Jian-Hong;Li, Guang-Xin
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.10
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    • pp.4203-4209
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    • 2015
  • Exosomes, membranous nanovesicles, naturally carry bio-macromolecules or miRNA and play impoetant roles in tumor pathogenesis. Here, we showed that macrophages cell-derived exosomes can function as vehicles to deliver exogenous miR-21 inhibitor into BGC-823 gastric cancer cells. Exosomes loaded with miR-21inhibitor significantly increased miR-21 levels in BGC-823, but miR-21inhibitor loaded in exosomes exerted an opposite effect. miRNA transfected with exosomes had less cellular toxicity to host cells compared to conventional transfection methods. The miR-21inhibitor loaded exosomes promoted the migration ability and reduced apoptosis of BGC-823 gastric cancer cells. These observations indicate that miR-21 acts as a tumor promoter by targeting the PDCD4 gene and preventing apoptosis of gastric cancer cells through inhibition of PDCD4 expression. Furthermore, exosome -mediated miR-21 inhibitor delivery resulted in functionally more efficient inhibition and less cellular toxicity compared to conventional transfection methods. Similar approaches could be useful in modification of target biomolecules in vitro and in vivo. These findings contribute to our understanding of the functions of miR-21 and exosomes as a carrier for therapy of gastric cancer.

Penetration of HEp-2 and Chinese Hamster Ovary Epithelial Cells by Escherichia coli Harbouring the Invasion-Conferring Genomic Region from Salmonella typhimurium

  • 박정욱;황상구;문자영;조용권;김동완;정용기
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.270-270
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    • 2002
  • Pathogenic Salmonella typhimurium can invade the intestinal epithelium and cause a wide range of diseases including gastroenteritis and bacteremia in human and animals. To identify the genes involved in the infection, the invasion determinant was obtained from S. typhimurium 82/6915 and was subcloned into pGEM-7Z. A subclone DHl (pSV6235) invaded HEp-2 and Chinese hamster ovary epithelial cells and contained a 4.4 kb fragment of S. typhimurium genomic region. Compared with the host strain E. coli DHl, the subclone DHl (pSV6235) invaded cultured HEp-2 and Chinese hamster ovary cells at least 75- and 68-fold higher, respectively. The invasion rate of E. coli DHl for the cells significantly increased by harbouring the genomic region derived from pathogenic S. typhimurium 82/6915.

Immunomodulation of Fungal β-Glucan in Host Defense Signaling by Dectin-1

  • Batbayar, Sainkhuu;Lee, Dong-Hee;Kim, Ha-Won
    • Biomolecules & Therapeutics
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    • v.20 no.5
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    • pp.433-445
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    • 2012
  • During the course of evolution, animals encountered the harmful effects of fungi, which are strong pathogens. Therefore, they have developed powerful mechanisms to protect themselves against these fungal invaders. ${\beta}$-Glucans are glucose polymers of a linear ${\beta}$(1,3)-glucan backbone with ${\beta}$(1,6)-linked side chains. The immunostimulatory and antitumor activities of ${\beta}$-glucans have been reported; however, their mechanisms have only begun to be elucidated. Fungal and particulate ${\beta}$-glucans, despite their large size, can be taken up by the M cells of Peyer's patches, and interact with macrophages or dendritic cells (DCs) and activate systemic immune responses to overcome the fungal infection. The sampled ${\beta}$-glucans function as pathogen-associated molecular patterns (PAMPs) and are recognized by pattern recognition receptors (PRRs) on innate immune cells. Dectin-1 receptor systems have been incorporated as the PRRs of ${\beta}$-glucans in the innate immune cells of higher animal systems, which function on the front line against fungal infection, and have been exploited in cancer treatments to enhance systemic immune function. Dectin-1 on macrophages and DCs performs dual functions: internalization of ${\beta}$-glucan-containing particles and transmittance of its signals into the nucleus. This review will depict in detail how the physicochemical nature of ${\beta}$-glucan contributes to its immunostimulating effect in hosts and the potential uses of ${\beta}$-glucan by elucidating the dectin-1 signal transduction pathway. The elucidation of ${\beta}$-glucan and its signaling pathway will undoubtedly open a new research area on its potential therapeutic applications, including as immunostimulants for antifungal and anti-cancer regimens.

Subcellular Responses in Nonhost Plant Infected with Pathogenic and Non-pathogenic Strains of Xanthomonas axonopodis pv. glycines

  • Jeong, Yong-Ho;Kim, Jung-Gun;Chang, Sung-Pae;Hwang, In-Gyu;Kim, Young-Ho
    • The Plant Pathology Journal
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    • v.18 no.3
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    • pp.115-120
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    • 2002
  • Xanthomonas axonopodis pv. glycines, the causal agent of bacterial pustule of soybean, induces hypersensitive response (HR) in a non-host plant, hot pepper (Capsicum annuum). A wild-type strain (8ra) and its non-patho-genic mutant (8-13) of X. axonopodis pv. glycines were inoculated into the pepper leaf tissues and their subcellular responses to the bacterial infections were examined by electron microscopy. Intrastructural changes related to HR were found in the leaf tissues infected with 8ra from 8 h after inoculation, characterized by separation of plasmalemma from the cell wall, formation of small vacuoles and vesicles, formation of cell wall apposition, and cellular necrosis. No such responses were observed in the tissues infected with the mutant. In 8ra, the bacterial cells were attached to the cell walls, with the cell wall material dissolved into and appearing to encapsulate the bacterial cells. The bacterial cells later became entirely embedded in the cell wall material. On the other hand, in 8-13, the bacterial cells were usually not attached tightly to the plant cell wall, and no or poor encapsulation of the bacteria by the wall material occurred, although these were encircled by rather loose wall materials at the later stages.

Anti-inflammatory Effect of Mangosteen (Garcinia mangostana L.) Peel Extract and its Compounds in LPS-induced RAW264.7 Cells

  • Widowati, Wahyu;Darsono, Lusiana;Suherman, Jo;Fauziah, Nurul;Maesaroh, Maesaroh;Erawijantari, Pande Putu
    • Natural Product Sciences
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    • v.22 no.3
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    • pp.147-153
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    • 2016
  • Inflammation plays an important role in host defense against external stimuli such as infection by pathogen, endotoxin or chemical exposure by the production of the inflammatory mediators that produced by macrophage. Anti-inflammatory factor is important to treat the dangers of chronic inflammation associated with chronic disease. This research aims to analyze the anti-inflammatory effects of Garcinia mangostana L. peel extract (GMPE), ${\alpha}$-mangostin, and ${\gamma}$-mangostin in LPS-induced murine macrophage cell line (RAW 264.7) by inhibiting the production of inflammatory mediators. The cytotoxic assay of G. mangostana L. extract, ${\alpha}$-mangostin, and ${\gamma}$-mangostin were performed by MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) to determine the safe and non-toxic concentration in RAW 264.7 for the further assay. The concentration of inflammatory mediators (COX-2, IL-6, and IL-$1{\beta}$) were measured by the ELISA-based assay and NO by the nitrate/nitrite colorimetric assay in treated LPS-induced RAW 264.7 cells. The inhibitory activity was determined by the reducing concentration of inflammatory mediators in treated LPS-induced RAW 264.7 over the untreated cells. This research revealed that GMPE, ${\alpha}$-mangostin, and ${\gamma}$-mangostin possess the anti-inflammatory effect by reducing COX-2, IL-6, IL-$1{\beta}$, and NO production in LPS-induces RAW 264.7 cells.

CD7-Specific Single Chain Antibody Mediated Delivery of siRNA to T Cells Inhibits HIV Replication in a Humanized Mouse Model

  • Ban, Hong-Seok;Kumar, Priti;Kim, Na-Hyun;Choi, Chang-Son;Shankar, Premlata;Lee, Sang-Kyung
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2008.05a
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    • pp.62-64
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    • 2008
  • A major hurdle to the development of RNA interference as therapy for HIV infection is the delivery of siRNA to T lymphocytes which are difficult cells to transfect even in vitro. We have employed a single chain antibody to the pan T cell surface antigen CD7 was conjugated to an oligo-9-arginine peptide (scFvCD7-9R) for T cell-specific siRNA delivery in NOD/SCIDIL2${\gamma}$-/- mice reconstituted with human peripheral blood lymphocytes (Hu-PBL). Using a novel delivery, we first show that scFvCD7-9R efficiently delivered CD4 siRNA into human T cells in vitro. In vivo administration to Hu-PBL mice resulted in reduced levels of surface CD4 expression on T cells. Mice infected with HIV-1 and treated on a weekly basis with scFvCD7-9R-siRNA complexes targeting a combination of viral genes and the host coreceptor molecule CCR5 successfully maintained CD4/CD3 T cell ratios up to 4 weeks after infection in contrast to control mice that displayed a marked reduction in CD4 T cell numbers. p24 antigen levels were undetectable in 3 of the 4 protected mice. scFvCD7-9R/antiviral siRNA treatment also helped maintain CD4 T cell numbers with reduced plasma viral loads in Hu-PBL mice reconstituted with PBMC from donors seropositive for HIV, indicating that this method can contain viral replication even in established HIV infections. Our results show that scFvCD7-9R could be further developed as a potential therapeutic for HIV-1 infection.

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Expression patterns of innate immunity-related genes in response to polyinosinic:polycytidylic acid (poly[I:C]) stimulation in DF-1 chicken fibroblast cells

  • Jang, Hyun-Jun;Song, Ki-Duk
    • Journal of Animal Science and Technology
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    • v.62 no.3
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    • pp.385-395
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    • 2020
  • Polyinosinic:polycytidylic acid (poly[I:C]) can stimulate Toll-like receptor 3 (TLR3) signaling pathways. In this study, DF-1 cells were treated with poly(I:C) at various concentrations and time points to examine the comparative expression patterns of innate immune response genes. The viability of DF-1 cells decreased from 77.41% to 38.68% when cells were treated different dose of poly(I:C) from 0.1 ㎍/mL to 100 ㎍/mL for 24 h respectively. The expressions of TLR3, TLR4, TLR7, TLR15, TLR21, IL1B, and IL10 were increased in dose- and time-dependent manners by poly(I:C) treatment. On the contrary, the expression patterns of interferon regulatory factors 7 (IRF7), Jun proto-oncogene, AP-1 transcription factor subunit (JUN), Nuclear Factor Kappa B Subunit 1 (NF-κB1), and IL8L2 were varied; IRF7 and IL8L2 were increasingly expressed whereas the expressions of JUN and NF-κB1 were decreased in a dose-dependent manner after they were early induced. In time-dependent analysis, IRF7 expression was significantly upregulated from 3 h to 24 h, whereas JUN and NF-κB1 expressions settled down from 6 h to 24 h after poly(I:C) treatment although they were induced at early time from 1 h to 3 h. Poly(I:C) treatment rapidly increased the expression of IL8L2 from 3 h to 6 h with a plateau at 6 h and then the expression of IL8L2 was dramatically decreased until 24 h after poly(I:C) treatment although the expression level was still higher than the non-treated control. These results may provide the basis for understanding host response to viral infection and its mimicry system in chickens.

Effects of Chitosan on Anti-tumor Activity in Mice (키토산이 암세포성장에 미치는 효과)

  • 정양숙;김광혁;정영기;장명웅
    • Journal of Life Science
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    • v.14 no.2
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    • pp.209-214
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    • 2004
  • Cytotoxic anticancer chemotherapeutic agents generally produce severe side effects, while reducing host resistance to cancer and infections, especially through the destruction of lymphoid and bone marrow cells. In this study, we have investigated the effect of chitosan on cytotoxic activity against cancer cells and life span in mice. The direct cytotoxicity of chitosan or chitosan-combinated chemotherapeutic agents for tumor cells was observed. In addition, the effect of life span extention was counted on sarcoma 180 mice injected with chitosan-combinated mitomycin C. The effect of growth inhibion for cancer cells, K562 and Yac-1 was shown in the cytotoxicity test of chitosan or chitosan-combinated chemotherapeutic agents. Also, the effect of life span extension was observed on sarcoma 180 mice injected with chitosan-combinated mitomycin C. Our results suggest that life span extension in sarcom 180 mice exposed with chitosan-combinated chemotherapeutic agents showed the probability of its usefulness for cancer therapy if more research results were accumulated.

Gene Cloning and Characterization of MdeA, a Novel Multidrug Efflux Pump in Streptococcus mutans

  • Kim, Do Kyun;Kim, Kyoung Hoon;Cho, Eun Ji;Joo, Seoung-Je;Chung, Jung-Min;Son, Byoung Yil;Yum, Jong Hwa;Kim, Young-Man;Kwon, Hyun-Ju;Kim, Byung-Woo;Kim, Tae Hoon;Lee, Eun-Woo
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.430-435
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    • 2013
  • Multidrug resistance, especially multidrug efflux mechanisms that extrude structurally unrelated cytotoxic compounds from the cell by multidrug transporters, is a serious problem and one of the main reasons for the failure of therapeutic treatment of infections by pathogenic microorganisms as well as of cancer cells. Streptococcus mutans is considered one of the primary causative agents of dental caries and periodontal disease, which comprise the most common oral diseases. A fragment of chromosomal DNA from S. mutans KCTC3065 was cloned using Escherichia coli KAM32 as host cells lacking major multidrug efflux pumps. Although E. coli KAM32 cells were very sensitive to many antimicrobial agents, the transformed cells harboring a recombinant plasmid became resistant to several structurally unrelated antimicrobial agents such as tetracycline, kanamycin, rhodamin 6G, ampicillin, acriflavine, ethidium bromide, and tetraphenylphosphonium chloride. This suggested that the cloned DNA fragment carries a gene encoding a multidrug efflux pump. Among 49 of the multidrug-resistant transformants, we report the functional gene cloning and characterization of the function of one multidrug efflux pump, namely MdeA from S. mutans, which was expressed in E. coli KAM32. Judging from the structural and biochemical properties, we concluded that MdeA is the first cloned and characterized multidrug efflux pump using the proton motive force as the energy for efflux drugs.