scholarly journals APOPTOSIS STUDY OF INDONESIAN AVIAN INFLUENZA VIRUS SUBTYPE H5N1 IN MADIN-DARBY CANINE KIDNEY CELLS

2017 ◽  
Vol 11 (1) ◽  
pp. 39-44
Author(s):  
NLP Indi Dharmayanti ◽  
Dwi Rillah Ukhti ◽  
Farida Syamsiah ◽  
Risza Hartawan
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Avian Influenza Virus ◽  
Mdck Cells ◽  
Madin Darby Canine Kidney ◽  
Dna Ladder ◽  
Pathogenic Avian Influenza Virus ◽  
Virus Subtype ◽  
Darby Canine Kidney ◽  
Canine Kidney

This study aimed to determine the ability of highly pathogenic avian influenza virus (HPAI) virus subtype H5N1 originated from Indonesia to induce apoptosis in Madin-Darby Canine Kidney (MDCK) cells. Three HPAI virus subtype H5N1 isolates with different genetic characteristic namely A/Bird/Bali1/2011, A/Chicken/East Java/BwiI2/2010 and A/Chicken/West Java/1074/2003, were cultured in MDCK cells. Apoptosis was identified by deoxyribonucleic acid (DNA) fragmentation of infected MDCK cells using Apoptotic DNA Ladder Kit. The results showed that all three HPAI virus isolates used in this study did not able to induce apoptosis in the MDCK cells within 5 to 72 hours post infection.

scholarly journals The MAL Proteolipid Is Necessary for Normal Apical Transport and Accurate Sorting of the Influenza Virus Hemagglutinin in Madin-Darby Canine Kidney Cells

1999 ◽  
Vol 145 (1) ◽  
pp. 141-151 ◽  
Author(s):  
Rosa Puertollano ◽  
Fernando Martín-Belmonte ◽  
Jaime Millán ◽  
María del Carmen de Marco ◽  
Juan P. Albar ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Mdck Cells ◽  
Basolateral Membrane ◽  
Ectopic Expression ◽  
Apical Surface ◽  
Madin Darby Canine Kidney ◽  
Transport Pathway ◽  
Influenza Virus Hemagglutinin ◽  
Darby Canine Kidney ◽  
Canine Kidney

The MAL (MAL/VIP17) proteolipid is a nonglycosylated integral membrane protein expressed in a restricted pattern of cell types, including T lymphocytes, myelin-forming cells, and polarized epithelial cells. Transport of the influenza virus hemagglutinin (HA) to the apical surface of epithelial Madin-Darby canine kidney (MDCK) cells appears to be mediated by a pathway involving glycolipid- and cholesterol- enriched membranes (GEMs). In MDCK cells, MAL has been proposed previously as being an element of the protein machinery for the GEM-dependent apical transport pathway. Using an antisense oligonucleotide-based strategy and a newly generated monoclonal antibody to canine MAL, herein we have approached the effect of MAL depletion on HA transport in MDCK cells. We have found that MAL depletion diminishes the presence of HA in GEMs, reduces the rate of HA transport to the cell surface, inhibits the delivery of HA to the apical surface, and produces partial missorting of HA to the basolateral membrane. These effects were corrected by ectopic expression of MAL in MDCK cells whose endogenous MAL protein was depleted. Our results indicate that MAL is necessary for both normal apical transport and accurate sorting of HA.

scholarly journals Heterocyclic compounds based on 3-(4-bromophenyl) azo-5-phenyl-2(3H)-furanone: Anti-avian influenza virus (H5N1) activity / Heterociklički derivati 3-(4-bromfenil) azo-5-fenil-2(3H)-furanona: Djelovanje na virus ptičje gripe (H5N1)

2012 ◽  
Vol 62 (4) ◽  
pp. 593-606 ◽  
Author(s):  
Eman M. Flefel ◽  
Randa E. Abdel-Mageid ◽  
Waled A. Tantawy ◽  
Mohamed A. Ali ◽  
Abd El-Galil E. Amr
Keyword(s):  
Influenza Virus ◽  
Antiviral Activity ◽  
Avian Influenza ◽  
Avian Influenza Virus ◽  
Heterocyclic Compounds ◽  
Madin Darby Canine Kidney ◽  
Plaque Reduction Assay ◽  
Avian Influenza Virus H5n1 ◽  
Darby Canine Kidney

1 3-[2-(4-Bromphenyl)hydrazono]-5-phenyl-furan-2(3H)-one () was used for preparation of some novel pyrazole, pyridazinone, oxadiazole, triazole, thiazolidine and thioxopyrimidine derivatives. Some of the prepared products were tested for anti-avian influenza virus activity and revealed promising antiviral activity against H5N1 virus [A/Chicken/Egypt/1/20 % (H5N1)] by determination of both EC50 and LD50 and confirmed by plaque reduction assay on Madin-Darby canine kidney cells. Compounds 3-[2-(4-bromophenyl)hydrazono]-5-phenylfuran-2(3H)-one (1), 1-(4-bromophenyl)-N-hydroxy-5-phenyl-1H-pyrazole-3-carboxamide (5) and 1-(4-bromophenyl)-N-{2,3-dihydro-4-hydroxy-3-phenyl-6-oxo-2-thioxopyrimidin-1(6H)-yl}-5-phenyl-1H-pyrazole-3-carboxamide (12a) showed the highest effects. Detailed synthesis, spectroscopic data, and antiviral activity of the synthesized compounds are reported.

scholarly journals An Intact Dilysine-like Motif in the Carboxyl Terminus of MAL Is Required for Normal Apical Transport of the Influenza Virus Hemagglutinin Cargo Protein in Epithelial Madin-Darby Canine Kidney Cells

2001 ◽  
Vol 12 (6) ◽  
pp. 1869-1883 ◽  
Author(s):  
Rosa Puertollano ◽  
José Angel Martı́nez-Menárguez ◽  
Alicia Batista ◽  
José Ballesta ◽  
Miguel Angel Alonso
Keyword(s):  
Influenza Virus ◽  
Mdck Cells ◽  
Protein Sorting ◽  
Apical Surface ◽  
Carboxy Terminus ◽  
Madin Darby Canine Kidney ◽  
Influenza Virus Hemagglutinin ◽  
Carboxyl Terminal ◽  
Darby Canine Kidney ◽  
Canine Kidney

The MAL proteolipid, a component of the integral protein sorting machinery, has been demonstrated as being necessary for normal apical transport of the influenza virus hemagglutinin (HA) and the overall apical membrane proteins in Madin-Darby canine kidney (MDCK) cells. The MAL carboxy terminus ends with the sequence Arg-Trp-Lys-Ser-Ser (RWKSS), which resembles dilysine-based motifs involved in protein sorting. To investigate whether the RWKSS pentapeptide plays a role in modulating the distribution of MAL and/or its function in apical transport, we have expressed MAL proteins with distinct carboxy terminus in MDCK cells whose apical transport was impaired by depletion of endogenous MAL. Apical transport of HA was restored to normal levels by expression of MAL with an intact but not with modified carboxyl terminal sequences bearing mutations that impair the functioning of dilysine-based sorting signals, although all the MAL proteins analyzed incorporated efficiently into lipid rafts. Ultrastructural analysis indicated that compared with MAL bearing an intact RWKSS sequence, a mutant with lysine −3 substituted by serine showed a twofold increased presence in clathrin-coated cytoplasmic structures and a reduced expression on the plasma membrane. These results indicate that the carboxyl-terminal RWKSS sequence modulates the distribution of MAL in clathrin-coated elements and is necessary for HA transport to the apical surface.

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