• Title/Summary/Keyword: virus like particle

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Influenza M1 Virus-Like Particles Consisting of Toxoplasma gondii Rhoptry Protein 4

  • Lee, Su-Hwa;Lee, Dong-Hun;Piao, Ying;Moon, Eun-Kyung;Quan, Fu-Shi
    • Parasites, Hosts and Diseases
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    • v.55 no.2
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    • pp.143-148
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    • 2017
  • Toxoplasma gondii infections occur throughout the world, and efforts are needed to develop various vaccine candidates expressing recombinant protein antigens. In this study, influenza matrix protein (M1) virus-like particles (VLPs) consisting of T. gondii rhoptry antigen 4 (ROP4 protein) were generated using baculovirus (rBV) expression system. Recombinant ROP4 protein with influenza M1 were cloned and expressed in rBV. SF9 insect cells were coinfected with recombinant rBVs expressing T. gondii ROP4 and influenza M1. As the results, influenza M1 VLPs showed spherical shapes, and T. gondii ROP4 protein exhibited as spikes on VLP surface under transmission electron microscopy (TEM). The M1 VLPs resemble virions in morphology and size. We found that M1 VLPs reacted with antibody from T. gondii-infected mice by western blot and ELISA. This study demonstrated that T. gondii ROP4 protein can be expressed on the surface of influenza M1 VLPs and the M1 VLPs containing T. gondii ROP4 reacted with T. gondii-infected sera, indicating the possibility that M1 VLPs could be used as a coating antigen for diagnostic and/or vaccine candidate against T. gondii infection.

Production of Recombinant Human Papillomavirus Type 52 L1 Protein in Hansenula polymorpha Formed Virus-Like Particles

  • Liu, Cunbao;Yao, Yufeng;Yang, Xu;Bai, Hongmei;Huang, Weiwei;Xia, Ye;Ma, Yanbing
    • Journal of Microbiology and Biotechnology
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    • v.25 no.6
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    • pp.936-940
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    • 2015
  • Human papillomavirus (HPV) type 52 is a high-risk HPV responsible for cervical cancer. HPV type 52 is common around the world and is the most common in some Asian regions. The available prophylactic HPV vaccines protect only from HPV types 16 and 18. Supplementing economical vaccines that target HPV type 52 may satisfactorily complement available prophylactic vaccines. A codon-adapted HPV 52 L1 gene was expressed in the methylotrophic yeast Hansenula polymorpha, which is used as an industrial platform for economical hepatitis B surface antigen particle production in China. We found that the recombinant proteins produced in this expression system could form virus-like particles (VLPs) with diameters of approximately 50 nm. This study suggests that the HPV 52 VLPs produced in this platform may satisfactorily complement available prophylactic vaccines in fighting against HPVs prevalent in Asia.

Characterization of a Cell Line HFH-T2, Producing Viral Particles, from Primary Human Fetal Hepatocytes Infected with Hepatitis B Virus

  • Shim, Jae-Kyoung;Kim, Dong-Wook;Chung, Tae-Ho;Kim, June-Ki;Suh, Jeong-Ill;Park, Chun;Lee, Young-Choon;Chung, Tae-Wha;Song, Eun-Young;Kim, Cheorl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.186-192
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    • 2001
  • A primary culture of human fetal hepatocytes was obtained through a therapeutic abortion process at 26 weeks of gestation period. More than $10^8$ cells were seeded on a plastic plate. These hepatocytes were infected with hepatitis B virus (HBV). The HBV was purified from serum of one chronic HBV carrier. Transformed hepatocytes were subcultured in a 10% FBS-supplemented medium. The morphology of the transformed cell was epithelial-like. The cells from the first pass showed signs of early proliferation and had a latent period of more than 3 months after 6-7 passages. After the rest period, the transformed cell proliferated actively and they were subcultured every three days. Transformed hepatocytes were characterized by detection of the HBV transcript by RT-PCR. The secretion of virions from transformed cells was investigated by PCR with the cell medium. Two types of virions secreted into the culture medium were examined by using the transmission electron microscope. Another approach to study the secretion of virions in to culture medium was carried out with HBV antibody. HBsAg was detected in the culture medium of transformed cells using ELISA and Western blot analyses. These data suggested that the human fetal hepatocyte cell line has been established by infection of HBV, in which this cell line secreted viral particles into the culture medium.

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Expression and Characterization of Fusion Protein with Autographa californica Nuclear Polyhedrosis Virus Polyhedrin and Green Fluorescent Protein in Insect Cells (곤충세포주에서 Autographa californica 핵다각체병 바이러스의 다각체 단백질과 초록색 형광 단백질의 융합단백질 발현 및 특성)

  • 제연호;진병래;노종열;장진희;강석권
    • Korean journal of applied entomology
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    • v.38 no.2
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    • pp.139-144
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    • 1999
  • We have now constructed a novel recombinant baculovirus producing fusion protein with Autogrqha c.uliforrzica nuclear polyhedrosis virus (AcNPV) polyhedrin and green fluorescent protein (GFP). The fusion protein expressed by the recombinant baculovirus in insect cells was characterized. The GFP gene was introduced under the control of polyhedrin gene promoter of AcNPV, by fusion in the front or back of intact polyhedrin gene. The recombinant baculoviruses were named as Ac-GFPPOL or Ac-POLGFP. respectively. As expected, the 56 kDa fusion protein was expressed in the recombinant virus-infected cells. Interestingly. however, the fluorescence of GFP in the cells infected with Ac- POLGFP was only detected within the nuclei. and that was observed as polyhedra-like granular particles. In the microscopy of cells infected with Ac-GFPPOL, furthermore, GFP was detected in both cytoplasm and nuclei although most of GFP were present within the nuclei. However, fusion protein produced by recombinant virus did not form polyhedra although the fusion protein was fused with polyhedrin and GFP. It is suggested that difference of GFP location in the infected cells appear to be involved in the region of polyhedrin in the fusion protein, and the polyhedrin in the fusion protein might be responsible for the polyhedra-like granular particles present within nuclei.

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Ultrastructure of Cymbidium Leaf Tissue Systemically Infected with Odontoglossum Ringspot Virus (오돈토글로썸 윤문 바이러스에 감염된 Cymbidium 잎조직의 미세구조)

  • Ryu, Ki-Hyun;Kim, Woo-Kap;Park, Won-Mok
    • Applied Microscopy
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    • v.24 no.3
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    • pp.46-54
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    • 1994
  • In ultrathin section of the tissue of odontoglossum ringspot virus (ORSV)-infected Cymbidium goeringii Reichenbach, ORSV particles appeared as bundles of irregular aggregates of various length which were called stacked plates or rounded plates. Virus particles were found in the cytoplasm in electron clear zones and they also found between cell wall and plasma membrane. They mainly clustered in parallel aggregates and sometimes oriented randomly. The X-bodies and paramural bodies were observed near the cell membrane and these contained vacuole-like cavities. The cell wall of infected tissue expanded largely. Some chloroplast in ORSV infected cell was irregular. No virus particle was present in mitochondria, nuclei, vacuoles, vesicles or other organelles. The plasmodesmata slightly enlarged, and virus-associated granules were present around it.

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Hepatitis C Virus - Proteins, Diagnosis, Treatment and New Approaches for Vaccine Development

  • Keyvani, Hossein;Fazlalipour, Mehdi;Monavari, Seyed Hamid Reza;Mollaie, Hamid Reza
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.5917-5935
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    • 2012
  • Background: Hepatitis C virus (HCV) causes acute and chronic human hepatitis infection and as such is an important global health problem. The virus was discovered in the USA in 1989 and it is now known that three to four million people are infected every year, WHO estimating that 3 percent of the 7 billion people worldwide being chronically infected. Humans are the natural hosts of HCV and this virus can eventually lead to permanent liver damage and carcinoma. HCV is a member of the Flaviviridae family and Hepacivirus genus. The diameter of the virus is about 50-60 nm and the virion contains a single-stranded positive RNA approximately 10,000 nucleotides in length and consisting of one ORF which is encapsulated by an external lipid envelope and icosahedral capsid. HCV is a heterogeneous virus, classified into 6 genotypes and more than 50 subtypes. Because of the genome variability, nucleotide sequences of genotypes differ by approximately 31-34%, and by 20-23% among subtypes. Quasi-species of mixed virus populations provide a survival advantage for the virus to create multiple variant genomes and a high rate of generation of variants to allow rapid selection of mutants for new environmental conditions. Direct contact with infected blood and blood products, sexual relationships and availability of injectable drugs have had remarkable effects on HCV epidemiology. Hundreds of thousands of people die each year from hepatitis and liver cancer caused by HCV virus infection. Approximately 80% of patients with acute hepatitis C progress into a chronic disease state leading to serious hepatic disorders, 10-20% of which develop chronic liver cirrhosis and hepatocellular carcinoma. The incubation period of HCV is 6-8 weeks and the infection is often asymptomatic so it is very hard to detect at early stages, making early treatment very difficult. Therefore, hepatitis C is called a "silent disease". Neutralizing antibodies are produced against several HCV proteins during infection but the virus mutates to escape from antibodies. Some patients with chronic hepatitis C may have some symptoms such as fatigue, muscle aches, nausea and pain. Autoimmune and immunecomplex-mediated diseases have also been reported with chronic HCV infection.

Ultrastructural Aspects of Mixed Infections with Turnip mosaic virus (TuMV- ACl8 and -C5) and Ribgrass mosaic virus (RMV-CA1) in Oriental Cabbage

  • Cho, Jeom-Deog;Park, Hong-Soo;Kim, Jeong-Soo;La, Yong-Jun;Kim, Kyung-Soo
    • The Plant Pathology Journal
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    • v.18 no.4
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    • pp.192-198
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    • 2002
  • Mixed infections of two economically important viruses, Turnip mosaic virus(TuMV) in the family Potyviridae and Ribgrass mosaic virus(RMV) in the genus Tobamo-virus, were studied ultrastructurally on oriental cabbage. TuMV-ACl8 (alpine isolate in Korea) induced chlorotic spots on inoculated leaves of both ‘SSD63’ inbred line known as susceptible to TuMV, and ‘Tambok’ commercial cultivar, known as resistant to the virus, in the early stages of infection. TuMV-C5 (Taiwan isolate) caused severe mosaic and malformation on the upper leaves of ‘SSD63’, and necrotic spots in both inoculated and upper leaves of ‘Tambok’. RMV-CA1 (oriental cabbage isolate from alpine in Korea) induced vein chlorosis, leaf malformation, and midrib necrotic streak in the upper leaves of both ‘SSD63’ and ‘Tambok’. Both oriental cabbages infected with a combination of TuMV-ACl8 and RMV-CA1 showed synergistic symptoms of severe yellowing, severe mosaic, and necrotic spot or vein necrosis on their leaves. A combination of TuMV-C5 and RMV-CA1 produced synergistic symptoms only in ‘SSD63’. In ‘Tambok’ infected with the combination of TuMV-C5 and RMV-CA1, the number of necrotic spots on the inoculated leaves was one half lesser than that on singly infected with TuMV-C5. A few necrotic spots progressed systemically. In cells infected with a combination of TuMV-ACl8 and RMV-CA1 or TuMV-C5 and RMV-CA1, the particles of the two viruses made nonagon-like rings(NLR); one TuMV particle was surrounded loosely by nine RMV particles. Two unrelated viruses of TuMV and RMV were compacted in the central part of the spiral aggregates(SA) that was induced strikingly in cells by the mixed infections. The SA showed NLR in its center of the cross-sectioned side. Many particles of RMV of Tobamovirus were closely associated with Potyvirus-characteristic cylindrical inclusions. The SAs in the mixed infections were formed easily by the Potyvirus of TuMV-ACl8 or -C5 isolates.

Antiviral Potential of the Silkworm Deoxynojirimycin against Hepatitis B Virus

  • You, Jung-Eun;Seong, Su-Il;Kim, Young-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.7 no.2
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    • pp.139-144
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    • 2003
  • Over 350 million people worldwide are chronic carriers of hepatitis B virus (HBV). Chronic viral infections of the liver can progress to cirrhosis, which may ultimately lead to hepatic failure or the development of hepatocellular carcinoma. There are two antiviral drugs on the market approved for clinical management of chronic HBV infections; interferon-alpha and the nucleoside analog lamivudine. However, they showed adverse side-effects. In the rational drug design for such therapies we would like to utilize antiviral drugs that inhibit the HBV replication in the liver. Investigation of natural extracts of silkworm exhibiting antiviral potential was held in the functional HBV polymerase activity and the release of virion particle in the HepG2.2.15 cell lines. HBV-producing transgenic mouse fed with silkworm DNJ molecule was shown as an inhibitor of serum HBV particles. We could represent this DNJ molecule as an antiviral potential complementing conventional therapies after preclinical tests against WHBV-infected animal model, woodchuck.

Intracellular Signaling Pathway for Host Defense Mechanisms against Piscine Nervous Necrosis Virus (NNV) (어류신경괴사증바이러스(nervous necrosis virus, NNV) 감염에 따른 숙주의 방어기전관련 세포신호전달)

  • Kim, Jong-Oh
    • Journal of Life Science
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    • v.30 no.4
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    • pp.402-409
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    • 2020
  • Nervous necrosis virus (NNV) contains a bi-segmented viral genome, RNA1 (3.4 kb, RdRp), and RNA2 (1.4 kb, capsid protein) in a small particle (25 nm). Despite its extremely compact size, NNV has caused serious damage by infecting approximately 120 fish species worldwide since it was first reported in the late 1980s. In order to minimize the damage caused by NNV infection and develop effective vaccines, it is necessary to understand the intra cellular signaling system according to NNV infection. NNV infection induces cell cycle arrest at the G1 phase via the p53-dependent pathway to use the cellular system for its replication. Otherwise, host cells recognize NNV infection through the RIG-1-like receptor (RLR) signaling pathway to control the virus and infected cells, and then ISGs required for antiviral action are activated via the IFN signaling pathway. Moreover, apoptosis of infected cells is triggered by the unfolded protein response (UPR) through ER stress and mitochondria-mediated cell death. Cell signaling studies on the NNV infection mechanisms are still at an early stage and many pathways have yet to be identified. Understanding the various disease-specific cellular signaling systems associated with NNV infection is essential for rapid and accurate diagnosis and vaccine development.

Comparisons of Certification Standards for Mask and Review on Filtration Efficiency for Viruses (마스크의 인증기준 비교와 바이러스 여과효율에 대한 고찰)

  • Yoon, Chungsik;Go, Sulbee;Park, Jihoon
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.30 no.2
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    • pp.109-123
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    • 2020
  • Objectives: The aims of this study were to review the standards and key components of the standards for disposable masks in Korea, the US, EU, Japan, and China and to evaluate the appropriateness of disposable masks during a virus pandemic. Methods: We reviewed the standards in the above countries and compared their key elements for each standard. For the second purpose, systemic paper gathering using key words like 'mask', 'respirator' 'virus', and 'coronavirus' in the PubMed search engine was performed. Fifty-three papers were selected and reviewed in regard to the appropriateness of test protocols with sodium chloride(NaCl) particles for virus filtration and the effectiveness against viruses. Results: The standards for masks are largely divided into two categories: US standards and EU standards. In Korea, the Ministry of Employment and Labor adapted the EU standards for workers and the Health Masks adopted the Ministry of Employment and Labor standards by the Ministry of Food and Drug Safety. Regarding airborne viral infections, WHO emphasizes only droplet infection, while many studies have shown that small particles enter the air through coughing or sneezing, which increases the possibility of airborne infection. Compared to other particles, various factors such as airborne viability and the ability to replicate the virus in the body are further involved in the virus's airborne infection rate. Airborne infection is classified into absolute air infection, preferential air infection, and opportunistic air infection. The NaCl-certified N95 mask showed good filtration efficiency against viruses and NaCl particles were proved to be a surrogate material for viruses. From this, KF94 is also expected to be effective in blocking viruses. Conclusion: The N95 test method could be used as a surrogate test method for virus filtration. N95-class masks have been found to effectively block viral infections in the air. However, surgical or medical masks are only partially effective against airborne virus infection though they could effectively block large droplet infection. However, most studies considered in this study targeted N95 in foreign countries and studies on masks actually used in Korea are very limited, so studies on microorganisms and reuse on domestic masks should be conducted in the future.