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Temporal Morphogenesis of Herpes Simplex Virus Type 1-Infected and Brefeldin A-Treated Human Fibroblasts

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Abstract

Background

Insights in the herpesvirus-cell interactions are of general cell biology interest, especially to studies of intracellular transport, and of considerable significance in the efforts to generate drugs, vaccines, and gene therapy. However, the pathway of virus particle egress and maturation is a contentious issue.

Materials and Methods

The intracellular transport was inhibited in cultured herpes simplex virus type 1 (HSV-1) infected human fibroblasts by brefeldin A (BFA). The virus-cell interactions including the viral envelopment, transport of HSV-1 virions, and transport of viral glycoprotein D (gD-1) and glycoprotein C (gC-1) were studied by titration assay, immunoblot, immunofluorescence light microscopy, and immunogold electron microscopy of cryosections.

Results

gD-1 and gC-1 were synthesized and normally transported to the plasma membranes of untreated HSV-1 infected host cells. BFA (1 µg/ml medium) effectively blocked the transport of the glycoproteins to the plasma membranes and affected the tubulin and vimentin of the cytoskeleton. Viral particles and glycoproteins accumulated in the perinuclear space and the endoplasmic reticulum of BFA treated cells. Withdrawal of BFA influence up to 9 hr resulted in restored tubulin and vimentin, transport of glycoproteins to the plasma membranes, and steady release of infectious viral particles to the extracellular space superior to the cellular assembly of new virions. The ultrastructural data presented support that the primary envelopment of viral particles occur at the nuclear membranes containing immature glycoproteins followed by multiple de-envelopments and re-envelopments of the virions during the transport and maturation in the endoplasmic reticulum and the Golgi complex.

Conclusions

BFA-induced changes include the cytoskeleton with significant effect on HSV-1 maturation and egress. The data support a multiple-step envelopment of HSV-1 in a common pathway of glycoprotein synthesis and virion egress.

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Acknowledgments

This work was supported by grants from the Danish Cancer Society and by funds from the Health Insurance “danmark,” Frands Køhler Nielsen and wife, Erik Hørslev and wife, Birgit Hørslev, E. Danielsen and wife, Anna and Jakob Jakobsen, Valdemar and Thyra Foersom, Torben and Alice Frimodt, Aage Bang, and the LEO Foundation.

We are grateful to G.H. Cohen and R.J. Eisenberg (University of Pennsylvania, Philadelphia), S. Chatterjee (University of Alabama, Birmingham), and L. Pereira (University of California, San Francisco) for generous gifts of monoclonal antibodies. Cryosection and electron microscopy facilities were kindly made available by Professor O. Norén and Professor H. Sjöström (University of Copenhagen, Denmark).

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  1. The Protein Laboratory, Institute of Molecular Pathology, University of Copenhagen, The Panum Institute, Bldg. 6.2., Blegdamsvej 3C, DK-2200, Copenhagen N, Denmark

    Helle L. Jensen MD & Bodil Norrild

  2. The Department of Pathology, Gentofte University Hospital, Copenhagen, Denmark

    Helle L. Jensen MD

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Jensen, H.L., Norrild, B. Temporal Morphogenesis of Herpes Simplex Virus Type 1-Infected and Brefeldin A-Treated Human Fibroblasts. Mol Med 8, 210–224 (2002). https://doi.org/10.1007/BF03402013

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