• Title/Summary/Keyword: nucleoprotein

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Synthesis of New Uraci1-5-Sulphonamide-p-Phenyl Derivatives and Their Effect on Biomphalaria alexandrina Snail's Nucleoproteins

  • Fathalla, O.A.;Gad, H.S.M.;Maghaby, A.S.
    • Archives of Pharmacal Research
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    • v.23 no.2
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    • pp.128-138
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    • 2000
  • In continuation of the previous work (Fathalla, 1992) on the synthesis of some heterocycles containing uracil moiety, we report herein the incorporation of uracil moiety into cyan-opyridine thione, thiosemecarbazone, semicarbazone, cyanopyridine, ami nocyano pyridine, isoxazoline, pyrazoline, pyrimidine, triazolo pyrimidine, pyran, selena and thiazole derivatives which might modify their biological activities. The biological studies revealed that the chemical compound III f showed high molluscicdal activity than other compounds. The profile of the nucleoprotein extracted from chemically (compound IIIc, e, f and g) treated or UV-irradiated B.alexandrina snails did not show appreciable differences when compared to non-treated (native) snails by using SDS-PAGE, where no obvious qualitative or quantitative differences were observed. Immunization of experimental animals with the nucleoprotein extracted from native, chemically (compound III f & g) treated or physically treated B.alexandrina snails induced significant protection against challenge with normal S.mansoni cercariae, as compared to the non-immunized challenged control. As well as , a decrease in the number of granuloma formation and the size range of granuloma was also observed in immunized animals. It is concluded that, compounds III f and g have a potent molluscicidal activity. They also induced chemical modification comparable to that induced by physical treatment in the snail's nucleoprotein, which could possibly be used in immunization against S. mansoni infection.

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Improved Immune Response to Recombinant Influenza Nucleoprotein Formulated with ISCOMATRIX

  • Cargnelutti, Diego E.;Sanchez, Maria V.;Alvarez, Paula;Boado, Lorena;Glikmann, Graciela;Mattion, Nora;Scodeller, Eduardo A.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.416-421
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    • 2012
  • Current influenza vaccines elicit antibodies effective against homologous strains, but new strategies are urgently needed for protection against emerging epidemic or pandemic strains. Although influenza vaccine candidates based on the viral nucleoprotein (NP) or matrix protein do not elicit sterilizing immunity, they have the advantage of inducing immunity that may cover a larger number of viral strains. In this study, recombinant NP produced in Escherichia coli was purified and formulated in combination with the adjuvant ISCOMATRIX. This formulation increased a NP-specific immunity in mice, with a Th1 profile, and may constitute a promising low-cost influenza vaccine candidate, with ability to stimulate humoral and cellular immune responses.

Improving siRNA design targeting nucleoprotein gene as antiviral against the Indonesian H5N1 virus

  • Hartawan, Risza;Pujianto, Dwi Ari;Dharmayanti, Ni Luh Putu Indi;Soebandrio, Amin
    • Journal of Veterinary Science
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    • v.23 no.2
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    • pp.24.1-24.10
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    • 2022
  • Background: Small interfering RNA technology has been considered a prospective alternative antiviral treatment using gene silencing against influenza viruses with high mutations rates. On the other hand, there are no reports on its effectiveness against the highly pathogenic avian influenza H5N1 virus isolated from Indonesia. Objectives: The main objective of this study was to improve the siRNA design based on the nucleoprotein gene (siRNA-NP) for the Indonesian H5N1 virus. Methods: The effectiveness of these siRNA-NPs (NP672, NP1433, and NP1469) was analyzed in vitro in Marbin-Darby canine kidney cells. Results: The siRNA-NP672 caused the largest decrease in viral production and gene expression at 24, 48, and 72 h post-infection compared to the other siRNA-NPs. Moreover, three serial passages of the H5N1 virus in the presence of siRNA-NP672 did not induce any mutations within the nucleoprotein gene. Conclusions: These findings suggest that siRNA-NP672 can provide better protection against the Indonesian strain of the H5N1 virus.

A "Prime and Deploy" Strategy for Universal Influenza Vaccine Targeting Nucleoprotein Induces Lung-Resident Memory CD8 T cells

  • Haerynn Chung;Eun-Ah Kim;Jun Chang
    • IMMUNE NETWORK
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    • v.21 no.4
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    • pp.28.1-28.14
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    • 2021
  • Lung-resident memory T cells (TRM) play an essential role in protecting against pulmonary virus infection. Parenteral administration of DNA vaccine is generally not sufficient to induce lung CD8 TRM cells. This study investigates whether intramuscularly administered DNA vaccine expressing the nucleoprotein (NP) induces lung TRM cells and protects against the influenza B virus. The results show that DNA vaccination poorly generates lung TRM cells and massive secondary effector CD8 T cells entering the lungs after challenge infection do not offer sufficient protection. Nonetheless, intranasal administration of non-replicating adenovirus vector expressing no Ag following priming DNA vaccination deploys NP-specific CD8 TRM cells in the lungs, which subsequently offers complete protection. This novel 'prime and deploy' strategy could be a promising regimen for a universal influenza vaccine targeting the conserved NP Ag.

Development and Characterization of Monoclonal Antibodies against Nucleoprotein for Diagnosis of Influenza A Virus

  • Nguyen, Hong Phuong;Kwak, Chaewon;Heo, Chang-Kyu;Cho, Eun Wie;Yang, Jihyun;Poo, Haryoung
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.809-815
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    • 2018
  • Influenza, which is a highly contagious disease caused by the influenza A virus, continues to be a major health concern worldwide. Although the accurate and early diagnosis of influenza virus infection is important for controlling the spread of this disease and rapidly initiating antiviral therapy, the current influenza diagnostic kits are limited by their low sensitivity. In this study, we developed several new influenza nucleoprotein (NP)-specific monoclonal antibodies (mAbs) and compared their sensitivity and specificity of those with commercially available anti-NP mAbs. Three mAbs, designated M24.11, M34.3, and M34.33, exhibited higher reactivities to recombinant NPs and A/Puerto Rico/8/1934 (H1N1) viral lysates compared with the commercial mAbs, as assessed using enzyme-linked immunosorbent assays. M34.3 and M34.33 showed higher reactivities with A/California/04/09 (pandemic H1N1) and A/Philippines/2/82 (H3N2) viral lysates than the commercial mAbs. In contrast, M24.11 had marked reactivity with H3N2 but not with pandemic H1N1. Immunofluorescent confocal microscopy showed that the three mAbs effectively detected the presence of influenza virus in lung tissues of mice infected with A/Puerto Rico/8/1934. These results indicate that the newly developed M34.3 and M34.33 mAbs could be useful for the development of influenza diagnostics.

Optimized Expression, Purification, and Rapid Detection of Recombinant Influenza Nucleoproteins Expressed in Sf9 Insect Cells

  • Yoon, Sung-Jin;Park, Young-Jun;Kim, Hyun Ju;Jang, Jinwoo;Lee, Sang Jun;Koo, Sunwoo;Lee, Moo-Seung
    • Journal of Microbiology and Biotechnology
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    • v.28 no.10
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    • pp.1683-1690
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    • 2018
  • Accurate and rapid diagnosis of influenza infection is essential to enable early antiviral treatment and reduce the mortality associated with seasonal and epidemic infections. Immunochromatography is one of the most common methods used for the diagnosis of seasonal human influenza; however, it is less effective in diagnosing pandemic influenza virus. Currently, rapid diagnostic kits for pandemic influenza virus rely on the detection of nucleoprotein (NP) or hemagglutinin (HA). NP detection shows higher specificity and is more sensitive than HA detection. In this study, we time-dependently screened expression conditions, and herein report optimal conditions for the expression of recombinant nucleoprotein (rNP), which was 48 h after infection. In addition, we report the use of the expressed rNP in a rapid influenza diagnostic test (SGT i-flex Influenza A&B Test). We constructed expression vectors that synthesized rNP (antigen) of influenza A and B in insect cells (Sf9 cells), employed the purified rNP to the immunoassay test kit, and clearly distinguished NPs of influenza A and influenza B using this rapid influenza diagnostic kit. This approach may improve the development of rapid test kits for influenza using NP.

Molecular epidemiological study of measles virus throughout an imported epidemic outbreak in Gyeonggi-do in 2014 (2014년 경기지역에서 발생한 해외유입 홍역 바이러스의 분자역학적 특성 분석)

  • Yun, Hee-Jeong;Park, Po-Hyun;Hwang, Sun-Il;Huh, Jeong-Weon;Nam, Soo-Jung;Yong, Kum-Chan;Yoon, Mi-Hye
    • Korean Journal of Microbiology
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    • v.53 no.1
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    • pp.1-8
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    • 2017
  • Measles virus is a highly contagious, exanthematic virus, preventable by the use of an effective live-attenuated vaccine. However, measles virus remains endemic in many area of the world causing nearly 200,000 deaths per year and still a major cause of child mortality, mostly in developing countries. In March 2014, Republic of Korea was certified as a 'national measles elimination' by the WHO as a result of a high-quality case-based surveillance system and population immunity, which was achieved by a high vaccination rate (>95.0% since 1996). But, since the beginning of 2014, the Gyeonggi province has experienced a resurgence of measles cases. In this study, we investigated the characteristics of measles viruses isolated from confirmed measles in Gyeonggi province during January 1, 2014 ~ July 31, 2014, 60 isolates were obtained from 72 confirmed measles specimens. Genotypic distributions and genetic diversities of isolated measles virus were analyzed by sequencing of nucleoprotein (N) gene. 58 (96.7%) imported cases were identified. The predominant genotype was B3, which reflects the circulating measles virus in adjacent countries. The sequences of nucleoprotein (N) gene of isolated MeV were showed that the strains characterized showed the highest degree of identity (99%) with the Philippine related strains in 2013-2014. Therefore, infected traveler returning from the Philippines transmitted secondary infection in Korea.

Epigenetic aspects of telomeric chromatin in Arabidopsis thaliana

  • Lee, Won Kyung;Cho, Myeon Haeng
    • BMB Reports
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    • v.52 no.3
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    • pp.175-180
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    • 2019
  • Telomeres are nucleoprotein complexes at the physical ends of linear eukaryotic chromosomes. They protect the chromosome ends from various external attacks to avoid the loss of genetic information. Telomeres are maintained by cellular activities associated with telomerase and telomere-binding proteins. In addition, epigenetic regulators have pivotal roles in controlling the chromatin state at telomeres and subtelomeric regions, contributing to the maintenance of chromosomal homeostasis in yeast, animals, and plants. Here, we review the recent findings on chromatin modifications possibly associated with the dynamic states of telomeres in Arabidopsis thaliana.

Enhancement of DNA Vaccine-induced Immune Responses by Influenza Virus NP Gene

  • Choi, So-Young;Suh, You-Suk;Cho, Jae-Ho;Jin, Hyun-Tak;Chang, Jun;Sung, Young-Chul
    • IMMUNE NETWORK
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    • v.9 no.5
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    • pp.169-178
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    • 2009
  • DNA immunization induces B and T cell responses to various pathogens and tumors. However, these responses are known to be relatively weak and often transient. Thus, novel strategies are necessary for enhancing immune responses induced by DNA immunization. Here, we demonstrated that co-immunization of influenza virus nucleoprotein (NP) gene significantly enhances humoral and cell-mediated responses to codelivered antigens in mice. We also found that NP DNA coimmunization augments in vivo proliferation of adoptively transferred antigen-specific CD4 and CD8 T cells, which enhanced protective immunity against tumor challenge. Our results suggest that NP DNA can serve as a novel genetic adjuvant in cocktail DNA vaccination.

Multiple Functions of the Amino-terminal Domain of Bacteriophage Lambda Integrase: A New Member of Three-stranded $\beta-sheet$ DNA-binding Proteins

  • Cho Eun Hee
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.159-161
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    • 2002
  • Bacteriophage lambda integrase carries out the site-specific recombination of lambda. Integrase contains two DNA binding domains with distinct sequence specificity, namely arm-type binding and core-type binding domains. The amino-terminal arm-binding domain is structurally related to the three-stranded $\beta-sheet$ family of DNA-binding domains. Integrase binding to the high affinity arm-type site by the amino-terminal domain facilitates Int binding to the low affinity core-type site, where the cleavage and strand exchange occurs. The amino-terminal domain of Int also modulates the core-binding and catalysis through intramolecular domain-domain interaction and/or intermolecular interactions between Int monomers. In addition, the amino-terminal domain interacts cooperatively with excisionase during excision. This indicates that amino-terminal domain of Int plays an important role in formation of proper higher-order nucleoprotein structure required for lambda site-specific recombination.

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