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Gangliosides are not essential for influenza virus infection

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Abstract

Sialic acid is known to be an essential part of influenza virus receptors, but the specific identity of the receptor molecules on target cells is still not defined. In particular, the relative roles played by cellular sialylglycoproteins and gangliosides in virus entry into target cells remain unclear. To test whether gangliosides are essential for virus infection, we used the GM-95 mutant cell line of mouse B16 melanoma which lacks synthesis of major glycosphingolipids including gangliosides. We found that GM-95 cells grown in serum-containing medium harboured substantial amounts of ganglioside receptors for influenza virus due to incorporation of serum gangliosides. To obtain ganglioside-free cells, we adapted GM-95 cells to growth in defined serum-free (sf) medium. Ganglioside-free GM-95-sf cells could be infected by avian and human influenza A viruses and produced infectious virus progeny demonstrating that gangliosides were neither absolutely necessary for the early nor for the late stages of the infection. However, sensitivity of the GM-95-sf cells to the viruses was 2–4 times lower than that of the ganglioside-containing parent cell line. Further studies are needed to specify whether this effect was due to the lack of gangliosides, neutral glycosphingolipids, or other effects.

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Abbreviations

BSA:

bovine serum albumin

Dk/98:

A/Mallard duck/Alberta/279/98 (H3N8)

DMEM:

Dulbecco's modified Eagle medium

FCS:

fetal calf serum

GM-95-sf:

GM-95 cells adapted to serum-free medium

GD1a:

disialoganglioside Neu5Acα2-3Galβ1-3GalNAcβ1-4(Neu5Acα2-3)Galβ1-4Glc β1-1Cer

GM3:

monosialoganglioside Neu5Acα2-3Galβ1-4Glcβ1-1Cer

GT1b:

trisialoganglioside Neu5Acα2-3Galβ1-3GalNAcβ1-4(Neu5Acα2-8Neu5Acα2-3)Galβ1-4Glcβ1-1Cer

GalCer:

galactosylceramide

GlcCer:

glucosylceramide

HRP:

horseradish peroxidase

LacCer:

lactosylceramide

NP:

nucleoprotein of influenza virus

MDCK:

Madine-Darby canine kidney cells

PBS:

phosphate-buffered saline

TLC:

thin layer chromatography

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Authors and Affiliations

  1. Institute of Virology, Philipps University, Robert Koch str. 17, 35037, Marburg, Germany

    Mikhail Matrosovich

  2. M.P. Chumakov Institute of Poliomyelitis and Viral Encephalitides, Moscow, Russia

    Mikhail Matrosovich

  3. Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, Japan

    Takashi Suzuki

  4. Neuronal Circuit Mechanisms Research Group, Brain Science Institute, The Institute of Physical and Chemical Research (RIKEN), Japan

    Yoshio Hirabayashi

  5. Institute of Virology, Philipps University, Marburg, Germany

    Wolfgang Garten & Hans-Dieter Klenk

  6. Division of Virology, Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, TN, USA

    Robert G. Webster

Authors
  1. Mikhail Matrosovich
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  2. Takashi Suzuki
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  3. Yoshio Hirabayashi
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  4. Wolfgang Garten
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  5. Robert G. Webster
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  6. Hans-Dieter Klenk
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Correspondence to Mikhail Matrosovich.

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Matrosovich, M., Suzuki, T., Hirabayashi, Y. et al. Gangliosides are not essential for influenza virus infection. Glycoconj J 23, 107–113 (2006). https://doi.org/10.1007/s10719-006-5443-y

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