• Title/Summary/Keyword: cell infection

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Partial Characterization of the Pathogenic Factors Related to Chlamydia trachomatis Invasion of the McCoy Cell Membrane

  • Yeo, Myeng-Gu;Kim, Young-Ju;Park, Yeal
    • Journal of Microbiology
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    • v.41 no.2
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    • pp.137-143
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    • 2003
  • The present study was performed to identify pathogenic factors of Chlamydia trachomatis, which invade the host cell membrane. We prepared monoclonal antibody against C. trachomatis and searched for pathogenic factors using this antibody, and subsequently identified the surface components of the elementary body of C. trachomatis, i.e., major outer membrane protein (MOMP), lipopolysaccharide (LPS), and two other surface exposure proteins. These proteins are believed to be important in the pathogenesis of host cell chlamydial infection. Additionally, to identify factors related to the host cell and C. trachomatis, we prepared C. trachomatis infected and non-infected McCoy cell extracts, and reacted these with anti-chlamydial LPS monoclonal antibody. We found that anti-chlamydial LPS monoclonal antibody reacted with a 116 kDa proteinaceous McCoy cell membrane component.

Strategies for Transmissible Gastroenteritis Virus(TGEV) Vaccine Production by Swine testicle cells

  • Kim, Min-Young;Jeong, Yeon-Ho;Kim, Myoung-Hwa;Ko, Yun-Mi
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.203-206
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    • 2005
  • TGE (Transmissible Gastroenteritis) caused by a virus belonging to family coronavirus, results in an acute infection of the small intestine of the pig. The optimum operation variables such as multiplicity of infection (MOI), infection time and harvest time were investigated for TGE vaccine production by immobilized ST(swine testicle) cells. In the culture supplemented with 5% serum, maximum virus titer of $1.2{\times}10^6pfu/ml$ was obtained at the conditions of 0.01 MOI, 2day infection time, and 1 day harvest time. Serum is a potential source of bacterial, mycoplasmal and viral contamination, and it has a possibility of the introduction of serum proteins, prion and pyrogens into the final product. For these reasons, much attention has been focused on the development of serum-free media. A new serum-free media (SFM) has been developed in order to produce TGE vaccine of high quality with low cost. The performance of SFM developed was compared with other commercially available serum-free media and serum supplemented media in terms of virus productivity. The cultures with serum-free media showed higher titer than that with serum supplemented media. Among various serum-free media tested, CHO-S-SFMII showed highest virus titer.

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Age-Related $CD4^+CD25^+Foxp3^+$ Regulatory T-Cell Responses During Plasmodium berghei ANKA Infection in Mice Susceptible or Resistant to Cerebral Malaria

  • Shan, Ying;Liu, Jun;Pan, Yan-Yan;Jiang, Yong-Jun;Shang, Hong;Cao, Ya-Ming
    • Parasites, Hosts and Diseases
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    • v.51 no.3
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    • pp.289-295
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    • 2013
  • Different functions have been attributed to $CD4^+CD25^+Foxp3^+$ regulatory T-cells (Tregs) during malaria infection. Herein, we describe the disparity in Treg response and pro- and anti-inflammatory cytokines during infection with Plasmodium berghei ANKA between young (3-week-old) and middle-aged (8-month-old) C57BL/6 mice. Young mice were susceptible to cerebral malaria (CM), while the middle-aged mice were resistant to CM and succumbed to hyperparasitemia and severe anemia. The levels of pro-inflammatory cytokines, such as TNF-${\alpha}$, in young CM-susceptible mice were markedly higher than in middle-aged CM-resistant mice. An increased absolute number of Tregs 3-5 days post-inoculation, co-occurring with elevated IL-10 levels, was observed in middle-aged CM-resistant mice but not in young CM-susceptible mice. Our findings suggest that Treg proliferation might be associated with the suppression of excessive pro-inflammatory Th1 response during early malaria infection, leading to resistance to CM in the middle-aged mice, possibly in an IL-10-dependent manner.

Inhibitory Effect of Polysaccharide from Kefir Grain on the Infection of MA-104 Cell by Human Rotavirus (Kefir Grain의 Polysaccharide에 의한 HRV S-2의 MA-104 세포 감염억제)

  • Lee, Jong-Ik;Song, Kwang-Young;Chon, Jung-Whan;Hyeon, Ji-Yeon;Seo, Kun-Ho
    • Food Science of Animal Resources
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    • v.31 no.1
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    • pp.81-85
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    • 2011
  • The inhibitory effect of a polysaccharide from kefir on human rotavirus infection in MA-104 cells was investigated. The extracted polysaccharide was separated as fraction I in unbound materials and as fractions II, III, and IV in bound materials. Adding polysaccharide fractions II (4.8 mg/mL), III (5.3 mg/mL), and IV (1.4 mg/mL) inhibited the infection of MA-104 cells by human rotavirus. The 50% inhibitory concentrations (IC 50) were 0.075, 0.083, and 0.022 mg/mL, respectively. Based on these results, the kefir polysaccharide has anti-rotavirus activity. In conclusion, a polysaccharide from kefir had more than a 97% inhibition effect against human rotavirus infection in MA-104 cells.

Effect of Loranthi Ramuluswatet Extract on Macrophages Activation and Th1 Response (상기생 물추출물이 대식세포 활성화와 Th1 반응에 미치는 효과)

  • Shin, Hye-Young;Zhang, Wen-Ji;Kim, Youn-Chul;Yun, Yong-Gab;Park, Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.1
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    • pp.171-175
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    • 2008
  • In the recently, increased concern has been focused on the pharmacology and clinical utility of herbal extracts and derivatives as a drug or adjunct to chemotherapy and immunotherapy. Here we investigated the effect of the water extract of Loranthi Ramulus (LR) in production of inflammatory mediators and expression of toll-like receptor (TLR)-4, CD14 from peritoneal macrophage. We assayed the effect of LR water extract in cell proliferation in vitro and Th1/Th2 cytokine level in vivo. In peritoneal macrophages, water extract of LR water extract increased the production of Nitric oxide (NO) and $TNF-{\alpha}$. Also, LR water extract increased Con A-induced cell proliferation and IgG1, IgG2a level in serum. However, i.p. injection of water extract of LR water extract did not affect the level of $TNF-{\alpha}$, $IFN-{\gamma}$, IL-2, IL-4 and IL-5 in serum of mice. These studies indicate that LR water extract induces macrophage activation and suggest the possible use of LR water extract in macrophage-based immunotherapies.

Virus-like Particle Vaccine Containing Toxoplasma gondii Rhoptry Protein 13 Induces Protection against T. gondii ME49 Infection in Mice

  • Kang, Hae-Ji;Chu, Ki-Back;Lee, Su-Hwa;Kim, Min-Ju;Park, Hyunwoo;Jin, Hui;Quan, Fu-Shi
    • Parasites, Hosts and Diseases
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    • v.57 no.5
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    • pp.543-547
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    • 2019
  • Toxoplasma gondii can infect humans worldwide, causing serious diseases in pregnant women and immunocompromised individuals. T. gondii rhoptry protein 13 (ROP13) is known as one of the key proteins involved in host cell invasion. In this study, we generated virus-like particles (VLPs) vaccine expressing T. gondii rhoptry ROP13 and investigated VLPs vaccine efficacy in mice. Mice immunized with ROP13 VLPs vaccine elicited significantly higher levels of T. gondii-specific IgG, IgG1, IgG2a, and IgA antibody responses following boost immunization and challenge infection, whereas antibody inductions were insignificant upon prime immunization. Differing immunization routes resulted in differing antibody induction, as intranasal immunization (IN) induced greater antibody responses than intramuscular immunization (IM) after boost and challenge infection. IN immunization induced significantly higher levels of IgG and IgA antibody responses from feces, antibody-secreting cells (ASCs), $CD4^+$ T, $CD8^+$ T cells and germinal center B cell responses in the spleen compared to IM immunization. Compared to IM immunization, IN immunization resulted in significantly reduced cyst counts in the brain as well as lesser body weight loss, which contributed to better protection. All of the mice immunized through either route survived, whereas all na?ve control mice perished. These results indicate that the ROP13 VLPs vaccine could be a potential vaccine candidate against T. gondii infection.

Multifactorial Traits of SARS-CoV-2 Cell Entry Related to Diverse Host Proteases and Proteins

  • You, Jaehwan;Seok, Jong Hyeon;Joo, Myungsoo;Bae, Joon-Yong;Kim, Jin Il;Park, Man-Seong;Kim, Kisoon
    • Biomolecules & Therapeutics
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    • v.29 no.3
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    • pp.249-262
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    • 2021
  • The most effective way to control newly emerging infectious disease, such as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, is to strengthen preventative or therapeutic public health strategies before the infection spreads worldwide. However, global health systems remain at the early stages in anticipating effective therapeutics or vaccines to combat the SARS-CoV-2 pandemic. While maintaining social distance is the most crucial metric to avoid spreading the virus, symptomatic therapy given to patients on the clinical manifestations helps save lives. The molecular properties of SARS-CoV-2 infection have been quickly elucidated, paving the way to therapeutics, vaccine development, and other medical interventions. Despite this progress, the detailed biomolecular mechanism of SARS-CoV-2 infection remains elusive. Given virus invasion of cells is a determining factor for virulence, understanding the viral entry process can be a mainstay in controlling newly emerged viruses. Since viral entry is mediated by selective cellular proteases or proteins associated with receptors, identification and functional analysis of these proteins could provide a way to disrupt virus propagation. This review comprehensively discusses cellular machinery necessary for SARS-CoV-2 infection. Understanding multifactorial traits of the virus entry will provide a substantial guide to facilitate antiviral drug development.

CD11b Deficiency Exacerbates Methicillin-Resistant Staphylococcus aureus-Induced Sepsis by Upregulating Inflammatory Responses of Macrophages

  • Hyunsub Sim;Daecheol Jeong;Hye-In Kim;Seongwon Pak;Bikash Thapa;Hyung-Joo Kwon;Keunwook Lee
    • IMMUNE NETWORK
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    • v.21 no.2
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    • pp.13.1-13.19
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    • 2021
  • Macrophages are important for the first line of defense against microbial pathogens. Integrin CD11b, which is encoded by Itgam, is expressed on the surface of macrophages and has been implicated in adhesion, migration, and cell-mediated cytotoxicity. However, the functional impact of CD11b on the inflammatory responses of macrophages upon microbial infection remains unclear. Here, we show that CD11b deficiency resulted in increased susceptibility to sepsis induced by methicillin-resistant Staphylococcus aureus (MRSA) infection by enhancing the pro-inflammatory activities of macrophages. Upon infection with MRSA, the mortality of Itgam knockout mice was significantly higher than that of control mice, which is associated with increased production of TNF-α and IL-6. In response to MRSA, both bone marrow-derived macrophages and peritoneal macrophages lacking CD11b produced elevated amounts of pro-inflammatory cytokines and nitric oxide. Moreover, CD11b deficiency upregulated IL-4-induced expression of anti-inflammatory mediators such as IL-10 and arginase-1, and an immunomodulatory function of macrophages to restrain T cell activation. Biochemical and confocal microscopy data revealed that CD11b deficiency augmented the activation of NF-κB signaling and phosphorylation of Akt, which promotes the functional activation of macrophages with pro-inflammatory and immunoregulatory phenotypes, respectively. Overall, our experimental evidence suggests that CD11b is a critical modulator of macrophages in response to microbial infection.

Modulated Gene Expression of Toxoplasma gondii Infected Retinal Pigment Epithelial Cell Line (ARPE-19) via PI3K/Akt or mTOR Signal Pathway

  • Zhou, Wei;Quan, Juan-Hua;Gao, Fei-Fei;Ismail, Hassan Ahmed Hassan Ahmed;Lee, Young-Ha;Cha, Guang-Ho
    • Parasites, Hosts and Diseases
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    • v.56 no.2
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    • pp.135-145
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    • 2018
  • Due to the critical location and physiological activities of the retinal pigment epithelial (RPE) cell, it is constantly subjected to contact with various infectious agents and inflammatory mediators. However, little is known about the signaling events in RPE involved in Toxoplasma gondii infection and development. The aim of the study is to screen the host mRNA transcriptional change of 3 inflammation-related gene categories, PI3K/Akt pathway regulatory components, blood vessel development factors and ROS regulators, to prove that PI3K/Akt or mTOR signaling pathway play an essential role in regulating the selected inflammation-related genes. The selected genes include PH domain and leucine- rich-repeat protein phosphatases (PHLPP), casein kinase2 (CK2), vascular endothelial growth factor (VEGF), pigment epithelium-derived factor (PEDF), glutamate-cysteine ligase (GCL), glutathione S-transferase (GST), and NAD(P)H: quinone oxidoreductase (NQO1). Using reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), we found that T. gondii up-regulates PHLPP2, $CK2{\beta}$, VEGF, GCL, GST and NQO1 gene expression levels, but down-regulates PHLPP1 and PEDF mRNA transcription levels. PI3K inhibition and mTOR inhibition by specific inhibitors showed that most of these host gene expression patterns were due to activation of PI3K/Akt or mTOR pathways with some exceptional cases. Taken together, our results reveal a new molecular mechanism of these gene expression change dependent on PI3K/Akt or mTOR pathways and highlight more systematical insight of how an intracellular T. gondii can manipulate host genes to avoid host defense.

Distribution of Oncogenic Human Papillomavirus Genotypes at High Grade Cervical Lesions above CIN 2 Grade with Histological Diagnosis

  • Kim, Geehyuk;Park, Sungyoung;Wang, Hye-young;Kim, Sunghyun;Park, Sangjung;Yu, Kwangmin;Lee, Boohyung;Ahn, Seung-Ju;Kim, Eun-Joong;Lee, Dongsup
    • Biomedical Science Letters
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    • v.22 no.2
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    • pp.37-45
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    • 2016
  • High risk human papillomavirus (HR-HPV) is major risk factor for uterine cervical cancer. There are approximately 15 types of HR-HPV. Liquid based cytology samples (116 samples) with high grade cervical lesions belonging to cervical intraepithelial neoplasia (CIN) 2, CIN 3, carcinoma in situ (CIS) and squamous cell carcinoma (SCC) were used after histologic confirmation. HR-HPV genotype assay was conducted using DNA chips. The HR-HPV infection rate was 81.9% with SCC samples showing the highest HR-HPV infection rate of 31%. CIN 3, CIS and CIN 2 showed infection rates of 25%, 16.4% and 9.5%, respectively. According to age with HR HPV infection rate, the 30~39 years-old group showed the highest infection rate by 92.3%. According to distribution with HR HPV genotyping, HPV 16 showed the highest infection rate by 42.3% whereas HPV 33 and HPV 58 showed infection rates of 11.7% and 10.8%, respectively. HPV 18 which is the second most common infected HPV genotype in the world showed 3.6%. Of the three most common oncogenic HR-HPV genotypes in CIN 2, we detected HPV 16, 35, 58; CIN 3 was HPV 16, 33, 58; CIS was HPV 16, 58, 33 (35/52); and SCC was HPV 16, 33, and 18 (31/52/58). Among the HPV 18, CIN 2, CIN 3, CIS and SCC showed 0.9%, 0.9%, 0% and 1.8%, respectively. The most often used preventive vaccines for cervical cancers use HPV 16 and HPV 18 as targets. However, results derived from this study suggest that a preventive vaccine against HPV 16 and HPV 18 would not be optimal for populations in this study.