• Title/Summary/Keyword: cell infection

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Production of Recombinant Polyhedra Containing Cry1Ac Fusion Protein in Insect Cell Lines

  • Kim, Jae-Su;Choi, Jae-Young;Roh, Jong-Yul;Lee, Han-Young;Jang, Seung-Sik;Je, Yeon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.739-744
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    • 2007
  • Insect cell lines and the control of infection for obtaining the maximum amount of polyhedrin-Cry1Ac-polyhedrin fusion protein from Bactrus in monolayer and suspension culture systems were tested. Growth rates of the Trichoplusia ni(High-Five) cell line in both culture systems were better than the other insect cell lines, Spodoptera frugiferda(Sf-9, Sf-21), Trichoplusia ni(Tn5), and Spodoptera exigua(Se301). The expression of the fusion protein in a monolayer culture showed that Se301 cells were 2.3-4.8 times more productive on a per cell basis than the other cell lines. However, in suspension culture, only High-Five cells were productive. High-Five cells infected with Bactrus at a multiplicity of infection(MOI) of 5 and a cell density of $3.0{\times}10^5$ cells per ml were more productive than the other infection condition in a suspension culture suitable for a large-scale production of baculovirus. In conclusion, for the large-scale production of Bactrus in vitro, High-Five cells showing good growth and high productivity are suitable.

Genetic Studies on Faecal Egg Counts and Packed Cell Volume Following Natural Haemonchus contortus Infection and Their Relationships with Liveweight in Muzaffarnagari Sheep

  • Yadav, N.K.;Mandal, Ajoy;Sharma, D.K.;Rout, P.K.;Roy, R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.11
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    • pp.1524-1528
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    • 2006
  • A total of 437 animals, comprising lambs aged between 3 and 12 months and adults of either sex of Muzaffarnagari sheep maintained at the Central Institute for Research on Goats, Makhdoom, Farah, Mathura, India were screened to assess the prevalence of Haemonchus contortus infection following natural infection and to identify the various factors affecting faecal egg count (FEC) and packed cell volume (PCV) of ewes and their genetic control. The relationships between FEC, PCV and body weight were also estimated. The prevalence rate for H. contortus infection in the flock under study was 15.7% indicating much lower occurrence of worm infection in lambs up to one year of age. On the other hand, a large proportion i.e., 67.7% of sheep was refractive to natural H. contortus infection. The random effect of sire significantly contributed (p<0.01) variation in log-transformed FEC (LFEC) of ewes. The season of birth had a significant (p<0.01) effect on LFEC of ewes. The lactating ewes had significantly (p<0.01) higher faecal egg counts compared to dry and pregnant ewes. The linear regression effects of the age of ewes on LFEC of animals were significant (p<0.01) in the present study. The heritabilities of LFEC, PCV and body weights of ewes during the course of infection were moderate to high in magnitude and ranged from 0.24 to 0.47. The LFEC of ewes was significantly (p<0.05) and negatively correlated with PCV at both genetic and phenotypic level. The genetic and phenotypic relationships between LFEC and body weights of ewes were -0.26 and -0.06 for this breed. The genetic correlation of PCV and body weight of ewes was positive and high (0.58) and statistically significant (p<0.05) but it was negatively correlated (-0.01) with body weight at the phenotypic level.

Interaction between host cell proteins and open reading frames of porcine circovirus type 2

  • Si-Won Park;In-Byung Park;Seok-Jin Kang;Joonbeom Bae;Taehoon Chun
    • Journal of Animal Science and Technology
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    • v.65 no.4
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    • pp.698-719
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    • 2023
  • Postweaning multisystemic wasting syndrome (PMWS) is caused by a systemic inflammation after porcine circovirus type 2 (PCV2) infection. It was one of the most economically important pathogens affecting pig production worldwide before PCV2 vaccine was first introduced in 2006. After the development of a vaccine against PCV2a type, pig farms gradually restored enormous economic losses from PMWS. However, vaccine against PCV2a type could not be fully effective against several different PCV2 genotypes (PCV2b - PCV2h). In addition, PCV2a vaccine itself could generate antigenic drift of PCV2 capsid. Therefore, PCV2 infection still threats pig industry worldwide. PCV2 infection was initially found in local tissues including reproductive, respiratory, and digestive tracks. However, PCV2 infection often leads to a systemic inflammation which can cause severe immunosuppression by depleting peripheral lymphocytes in secondary lymphoid tissues. Subsequently, a secondary infection with other microorganisms can cause PMWS. Eleven putative open reading frames (ORFs) have been predicted to encode PCV2 genome. Among them, gene products of six ORFs from ORF1 to ORF6 have been identified and characterized to estimate its functional role during PCV2 infection. Acquiring knowledge about the specific interaction between each PCV2 ORF protein and host protein might be a key to develop preventive or therapeutic tools to control PCV2 infection. In this article, we reviewed current understanding of how each ORF of PCV2 manipulates host cell signaling related to immune suppression caused by PCV2.

Cytomegalovirus Infection and Memory T Cell Inflation

  • Kim, Jihye;Kim, A-Reum;Shin, Eui-Cheol
    • IMMUNE NETWORK
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    • v.15 no.4
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    • pp.186-190
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    • 2015
  • Cytomegalovirus (CMV) infection in healthy individuals is usually asymptomatic and results in latent infection. CMV reactivation occasionally occurs in healthy individuals according to their immune status over time. T cell responses to CMV are restricted to a limited number of immunodominant epitopes, as compared to responses to other chronic or persistent viruses. This response results in progressive, prolonged expansion of CMV-specific $CD8^+$ T cells, termed 'memory inflation'. The expanded CMV-specific $CD8^+$ T cell population is extraordinarily large and is more prominent in the elderly. CMV-specific $CD8^+$ T cells possess rather similar phenotypic and functional features to those of replicative senescent T cells. In this review, we discuss the general features of CMV-specific inflationary memory T cells and the factors involved in memory inflation.

Alterations of Protein Expression in Macrophages in Response to Candida albicans Infection

  • Shin, Yu-Kyong;Kim, Ki-Young;Paik, Young-Ki
    • Molecules and Cells
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    • v.20 no.2
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    • pp.271-279
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    • 2005
  • Although macrophages are an important first line of cellular defense, they are unable to effectively kill phagocytosed C. albicans. To determine the physiological basis of this inability, we investigated the alterations of macrophage proteins caused by C. albicans infection. Since the formation of C. albicans hyphae caused cell death, proteins were prepared 3 h after infection and examined by two-dimensional gel electrophoresis (2-DE). The most prominent changes were in glycolytic enzymes, which could have caused energy depletion of the infected cells. Also changed were proteins involved in maintenance of cellular integrity and NO production. Treatment of the macrophages with either cytochalasin D or taxol did not alter their inability to kill C. albicans. Our results indicate that multiple factors contribute to cell death as the pathogenic form of C. albicans becomes fully active inside macrophage cells.

GLOBAL STABILITY OF VIRUS DYNAMICS MODEL WITH IMMUNE RESPONSE, CELLULAR INFECTION AND HOLLING TYPE-II

  • ELAIW, A.M.;GHALEB, SH.A.
    • Journal of the Korean Society for Industrial and Applied Mathematics
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    • v.23 no.1
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    • pp.39-63
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    • 2019
  • In this paper, we study the effect of Cytotoxic T Lymphocyte (CTL) and antibody immune responses on the virus dynamics with both virus-to-cell and cell-to-cell transmissions. The infection rate is given by Holling type-II. We first show that the model is biologically acceptable by showing that the solutions of the model are nonnegative and bounded. We find the equilibria of the model and investigate their global stability analysis. We derive five threshold parameters which fully determine the existence and stability of the five equilibria of the model. The global stability of all equilibria of the model is proven using Lyapunov method and applying LaSalle's invariance principle. To support our theoretical results we have performed some numerical simulations for the model. The results show the CTL and antibody immune response can control the disease progression.

Effects of Recombinant Baculovirus Infection Conditions on Production of Green Fluorescent Protein in Drosophila S2 Cells (초파리 S2 세포 시스템에서 녹색형광단백질 생산을 위한 재조합 배큘로바이러스의 감염조건들의 영향)

  • Cho, Hye Sook;Kim, Yeon Kyu;Kim, Kyoung Ro;Cha, Hyung Joon
    • Korean Chemical Engineering Research
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    • v.44 no.1
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    • pp.40-45
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    • 2006
  • The baculovirus-insect Drosophila melanogaster S2 cell system combines advantages of conventional baculovirus system and non-lytic S2 cell system because baculoviruses can infect non-permissive cells such as mammalian and Drosophila S2 cells but cannot replicate themselves inside the cells. In the present work, we investigated effects of infection conditions on production of green fluorescent protein (GFP) as a target protein using this baculovirus-S2 cell system. Even though higher MOI and longer baculovirus contact time showed better GFP expression yield during the shorter period, overall protein yield could be lower during the longer period due to the relatively higher cell detachment and lysis (lower cell viability). In addition, maintaining high MOI will be not practical for large-scale cell culture. Therefore, instead of maintaining high MOI, we found that high initial cell number and concentrated (10X) baculovirus volume can confer comparable protein expression even under the moderate MOI condition. Also, we found that the post-infection time that is connected to state of cells after infection was an important factor for production yield.

Immune Responses to Viral Infection (바이러스 감염에 대한 면역반응)

  • Hwang, Eung-Soo;Park, Chung-Gyu;Cha, Chang-Yong
    • IMMUNE NETWORK
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    • v.4 no.2
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    • pp.73-80
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    • 2004
  • Viruses are obligate intracellular parasites which cause infection by invading and replicating within cells. The immune system has mechanisms which can attack the virus in extracellular and intracellular phase of life cycle, and which involve both non-specific and specific effectors. The survival of viruses depends on the survival of their hosts, and therefore the immune system and viruses have evolved together. Immune responses to viral infection may be variable depending on the site of infection, the mechanism of cell-to-cell spread of virus, physiology of the host, host genetic variation, and environmental condition. Viral infection of cells directly stimulates the production of interferons and they induce antiviral state in the surrounding cells. Complement system is also involved in the elimination of viruses and establishes the first line of defence with other non-specific immunity. During the course of viral infection, antibody is most effective at an early stage, especially before the virus enters its target cells. The virus- specific cytotoxic T lymphocytes are the principal effector cells in clearing established viral infections. But many viruses have resistant mechanism to host immune responses in every step of viral infection to cells. Some viruses have immune evasion mechanism and establish latency or persistency indefinitely. Furthermore antibodies to some viruses can enhance the disease by the second infection. Immune responses to viral infection are very different from those to bacterial infection.

Transcriptional Analysis of 10 Selected Genes in a Model of Penicillin G Induced Persistence of Chlamydophila psittaci in HeLa Cells

  • Hu, Yanqun;Chen, Lili;Wang, Chuan;Xie, Yafeng;Chen, Zhixi;Liu, Liangzhuan;Su, Zehong;Wu, Yimou
    • Journal of Microbiology and Biotechnology
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    • v.25 no.8
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    • pp.1246-1256
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    • 2015
  • Chlamydophila psittaci is an important intracellular pathogen. Persistent infection is an important state of the host-parasite interaction in this chlamydial infection, which plays a significant role in spreading the organism within animal populations and in causing chronic chlamydiosis and serious sequelae. In this study, a C. psittaci persistent infection cell model was induced by penicillin G, and real-time quantitative PCR was used to study the transcriptional levels of 10 C. psittaci genes (dnaA, dnaK, ftsW, ftsY, grpE, rpsD, incC, omcB, CPSIT_0846, and CPSIT_0042) in acute and penicillin-G-induced persistent infection cultures. Compared with the acute cultures, the penicillin-G-treated cultures showed a reduced chlamydial inclusion size and a significantly decreased number of elementary body particles. Additionally, some enlarged aberrant reticulate body particles were present in the penicillin-G-treated cultures but not the acute ones. The expression levels of genes encoding products for cell division (FtsW, FtsY) and outer membrane protein E encoding gene (CPSIT_0042) were downregulated (p < 0.05) from 6 h post-infection onward in the persistent infection cultures. Also from 6 h post-infection, the expression levels of DnaA, DnaK, IncC, RpsD, GrpE, and CPSIT_0846 were upregulated (p < 0.05); however, the expression level of OmcB in the persistent infection was< almost the same as that in the acute infection (p > 0.05). These results provide new insight regarding molecular activities that accompany persistence of C. psittaci, which may play important roles in the pathogenesis of C. psittaci infection.

Trichomonas vaginalis Metalloproteinase Induces mTOR Cleavage of SiHa Cells

  • Quan, Juan-Hua;Choi, In-Wook;Yang, Jung-Bo;Zhou, Wei;Cha, Guang-Ho;Zhou, Yu;Ryu, Jae-Sook;Lee, Young-Ha
    • Parasites, Hosts and Diseases
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    • v.52 no.6
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    • pp.595-603
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    • 2014
  • Trichomonas vaginalis secretes a number of proteases which are suspected to be the cause of pathogenesis; however, little is understood how they manipulate host cells. The mammalian target of rapamycin (mTOR) regulates cell growth, cell proliferation, cell motility, cell survival, protein synthesis, and transcription. We detected various types of metalloproteinases including GP63 protein from T. vaginalis trophozoites, and T. vaginalis GP63 metalloproteinase was confirmed by sequencing and western blot. When SiHa cells were stimulated with live T. vaginalis, T. vaginalis excretory-secretory products (ESP) or T. vaginalis lysate, live T. vaginalis and T. vaginalis ESP induced the mTOR cleavage in both time-and parasite load-dependent manner, but T. vaginalis lysate did not. Pretreatment of T. vaginalis with a metalloproteinase inhibitor, 1,10-phenanthroline, completely disappeared the mTOR cleavage in SiHa cells. Collectively, T. vaginalis metallopeptidase induces host cell mTOR cleavage, which may be related to survival of the parasite.