Hostname: page-component-76fb5796d-r6qrq Total loading time: 0 Render date: 2024-04-27T02:09:56.453Z Has data issue: false hasContentIssue false
  • English
  • Français

Influenza virus A subtype H1N1 is inhibited by methylated β-lactoglobulin

Published online by Cambridge University Press:  08 September 2010

Mahmoud Sitohy ,
Michela Scanu ,
Bernard Besse ,
Claudine Mollat ,
Sylviane Billaudel ,
Thomas Haertlé  and
Jean-Marc Chobert
Mahmoud Sitohy
Affiliation:
UR 1268 Biopolymères Interactions Assemblages, INRA, F-44300 Nantes Zagazig University, Biochemistry Department, Faculty of Agriculture, Zagazig, Egypt
Michela Scanu
Affiliation:
UR 1268 Biopolymères Interactions Assemblages, INRA, F-44300 Nantes
Bernard Besse
Affiliation:
Université de Nantes, UFR des Sciences Pharmaceutiques, Bactériologie-Virologie. 1, rue Gaston Veil, B.P. 53508, 44035 Nantes Cedex 1, France
Claudine Mollat
Affiliation:
Université de Nantes, UFR des Sciences Pharmaceutiques, Bactériologie-Virologie. 1, rue Gaston Veil, B.P. 53508, 44035 Nantes Cedex 1, France
Sylviane Billaudel
Affiliation:
Université de Nantes, UFR des Sciences Pharmaceutiques, Bactériologie-Virologie. 1, rue Gaston Veil, B.P. 53508, 44035 Nantes Cedex 1, France
Thomas Haertlé
Affiliation:
UR 1268 Biopolymères Interactions Assemblages, INRA, F-44300 Nantes
Jean-Marc Chobert*
Affiliation:
UR 1268 Biopolymères Interactions Assemblages, INRA, F-44300 Nantes
*
*For correspondence; e-mail: chobert@nantes.inra.fr
Get access Rights & Permissions [Opens in a new window]

Abstract

Addition of methylated β-lactoglobulin (Met-BLG) in the medium of MDCK cell lines infected with influenza virus subtype H1N1 reduced hemagglutination activity (HA) in a concentration dependent manner. Antiviral activity of Met-BLG depended on its concentration, viral load, and duration of infection. Using 17 μg/ml of Met-BLG inhibited 50% of HA of H1N1 grown in MDCK cells at 1 MOI after 24 h incubation at 37°C and in 5% CO2. Extension of incubation time enhanced antiviral action since the same concentration of Met-BLG inhibited about 61% of viral activity after 48 h. This viral inhibition was accompanied by a protection of MDCK cells as observed by using neutral red or by direct microscope examination. Reduction of viral RNA replication upon the addition of Met-BLG (50 μg/ml) was observed by real time-PCR showing a reduction of viral log value of about 0·9. When viral stock solution was mixed with 25 μg/ml Met-BLG in absence of cell lines, the morphology and viability of virus particles were significantly affected as observed by electron microscopy, and the number of intact virus particles was reduced by roughly 65%.

Keywords

Influenza virusH1N1β-LactoglobulinEsterificationInhibition
Type
Research Article
Information
Journal of Dairy Research , Volume 77 , Issue 4 , November 2010 , pp. 411 - 418
Copyright
Copyright © Proprietors of Journal of Dairy Research 2010

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Baba, M, Schols, D, Pauwels, R, Balzarini, J & De Clercq, E 1988 Fuschin acid selectively inhibits human immunodeficiency virus (HIV) replication in vitro. Biochemical and Biophysical Research Communication 155 1404–1411CrossRefGoogle Scholar
Baccam, P, Beauchemin, C, Macken, CA, Hayden, FG & Perelson, AS 2006 Kinetics of influenza A virus infection in humans. Journal of Virology 80 7590–7599CrossRefGoogle ScholarPubMed
Balzarini, J, Mitsuya, H, De Clercq, E & Broder, S 1986 Aurintricarboxylic acid and Evans Blue represent two different classes of anionic compounds which selectively inhibit the cytopathogenicity of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus. Biochemical and Biophysical Research Communication 136 64–71CrossRefGoogle Scholar
Boom, R, Sol, CJ, Salimans, MM, Jansen, CL, Wertheim-van Dilen, PM & van der Noordaa, J 1990 Rapid and simple method for purification of nucleic acids. Journal of Clinical Microbiology 28 495–503CrossRefGoogle ScholarPubMed
Boulo, S, Akarsu, H, Ruigrok, RW & Baudin, F 2007 Nuclear traffic of influenza virus proteins and ribonucleoprotein complexes. Virus Research 124 12–21Google ScholarPubMed
Bourmakina, SV & Garci-Sastre, A 2005 The morphology and composition of influenza A virus particles are not affected by low levels of M1 and M2 proteins in infected cells. Journal of Virology 79 7926–7932Google Scholar
De Clercq, E 2009 The next ten stories on antiviral drug discovery (Part E): Advents, advances, and adventures. Medicinal Research Review. DOI 10.1002/med.20179CrossRefGoogle Scholar
Donald, HB & Isaacs, A 1954 Counts of influenza virus particles. Journal of General Microbiology 10 457–464CrossRefGoogle ScholarPubMed
Hariton-Gazal, E, Rosenbluh, J, Graessmann, A, Gilon, C & Loyter, A 2003 Direct translocation of histone molecules across cell membranes. Journal of Cell Science 116 4577–4586CrossRefGoogle ScholarPubMed
Horne, RW & Wildy, P 1979 An historical account of the development and applications of the negative staining technique to the electron microscopy of viruses. Journal of Microscopy 117 103–122Google Scholar
Huang, T-S, Palese, P & Krystal, M 1990 Determination of influenza virus proteins required for genomic replication. Journal of Virology 64 5669–5673CrossRefGoogle Scholar
Invitrogene life technologies 2003 SuperScript. III One-Step RT-PCR System with Platinum® Taq DNA Polymerase, Cat. No. 12574–018Google Scholar
Mailliart, P & Ribadeau Dumas, B 1988 Preparation of β-lactoglobulin and α-lactalbumin-free proteins from whey retentate by sodium chloride salting out at low pH. Journal of Food Science 53 743–752CrossRefGoogle Scholar
Mc Kimm-Breschkin, J, Sahasrabudhe, A, Bilck, T & McDonald, M 2001 Mechanism of resistance of influenza virus to neuraminidase inhibitors. International Congress Series 1219 855–861CrossRefGoogle Scholar
Mc Kimm-Breschkin, J, Trivedi, T, Hampson, A, Hay, A, Klimov, A, Tashiro, M, Hayden, F & Zambon, M 2003 Neuraminidase sequence analysis and susceptibilities of influenza virus clinical isolates to zanamivir and oseltamivir. Antimicrobial Agents and Chemotherapy 47 2264–2272CrossRefGoogle ScholarPubMed
Murayama, R, Harada, Y, Shibata, T, Kuroda, K, Hayakawa, S, Shimizu, K & Tanaka, T 2007 Influenza A virus non-structural protein 1 (NS1) interacts with cellular multifunctional nucleolin during infection. Biochemical and Biophysical Research Communication 362 880–885Google ScholarPubMed
Neumann, G, Noda, T, Kawaoka, Y 2009 Emergence and pandemic potential of swine-origin H1N1 influenza virus. Nature 459 931–939CrossRefGoogle ScholarPubMed
Peiris, JSM, Yu, WC, Leung, CW, Cheung, CY, Ng, WF, Nicholls, JM, Ng, TK, Chan, KH, Lai, ST, Lim, WL, Yuen, KY, Guan, Y 2004 Re-emergence of fatal human influenza A subtype H5N1 disease. The Lancet 363 617–619CrossRefGoogle ScholarPubMed
Poole, E, Elton, D, Medcalf, L & Digard, P 2004 Functional domains of influenza A virus PB2 protein: identification of NP- and PB1-binding sites. Virology 321 120–133CrossRefGoogle ScholarPubMed
Rachakonda, PS, Veit, M, Korte, T, Ludwig, K, Bottcher, C, Huang, Q, Schmidt, MFG & Herrmann, A 2007 The relevance for the stability of the influenza virus hemagglutinin. FASEB Journal 21 995–1002CrossRefGoogle ScholarPubMed
Sitohy, MZ, Chobert, J-M & Haertlé, T 2001 Simplified short-time method for the esterification of milk proteins. Milchwissenschaft 56 127–131Google Scholar
Sitohy, M, Chobert, J-M & Haertlé, T 2005 Esterified whey proteins can protect Lactococcus lactis against bacteriophage infection. Comparison with the effect of native basic proteins and L-polylysines. Journal of Agricultural and Food Chemistry 53 3727–3734CrossRefGoogle ScholarPubMed
Sitohy, M, Chobert, J-M, Karwowska, U, Gozdzicka-Jozefiak, A & Haertlé, T 2006 Inhibition of bacteriophage M13 replication with esterified milk proteins. Journal of Agricultural and Food Chemistry 54 3800–3806CrossRefGoogle ScholarPubMed
Sitohy, M, Billaudel, S, Haertlé, T & Chobert, J-M 2007 Antiviral activity of esterified α-lactalbumin and β-lactoglobulin against herpes simplex virus type 1. Comparison with the effect of acyclovir and L-polylysines. Journal of Agricultural and Food Chemistry 55 10214–10220Google ScholarPubMed
Sitohy, M, Chobert, J-M, Dalgalarrondo, M, Nowoczin, M, Besse, B, Billaudel, S & Haertlé, T 2008 The effect of bovine whey proteins on the ability of Poliovirus and Coxsackie virus to infect Vero cells cultures. International Dairy Journal 18 658–668CrossRefGoogle Scholar
Sitohy, M, Besse, B, Billaudel, S, Haertlé, T & Chobert, J-M 2010 Anti-viral action of methylated β-lactoglobulin on the human influenza virus A subtype H3N2. Probiotics and Antimicrobial Proteins 2 104–111CrossRefGoogle Scholar
Spackman, E, Senne, DA, Myers, TJ, Bulaga, LL, Garber, LP, Perdue, ML, Lohman, K, Daum, LT & Suaez, DL 2002 Development of a real-time reverse transcriptase PCR assay for Type A influenza virus and the avian H5 and H7 hemagglutinin subtypes. Journal of Clinical Microbiology 40 3256–3260CrossRefGoogle ScholarPubMed
Tirabassi, RS & Enquist, LW 1998 Role of envelop protein in gE endocytosis in the pseudorabies virus life cycle. Journal of Virology 72 4571–4579CrossRefGoogle ScholarPubMed
WHO 2009 (http://www.who.int/csr/disease/swineflu/notes/h1n1_antiviral_resistance_20090708/en/index.html)Google Scholar
Yamaguchi, M, Danev, R, Nishiyama, K, Sugawara, K & Nagayama, K 2008 Zernike phase contrast electron microscopy of ice-embedded influenza A virus. Journal of Structural Biology 162 271–276CrossRefGoogle ScholarPubMed