Cholesterol-depleted cells that are relatively permissive for Semliki Forest virus infection

Marianne T. Marquardt, Margaret Kielian

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Semliki Forest virus (SFV), an enveloped alphavirus, infects cells by endocytosis followed by low pH-triggered fusion of the virus and endocytic vesicle membranes. Progeny virus is released by budding from the cell plasma membrane. In vitro SFV fusion with artificial liposomes is triggered by low pH and is dependent on the presence of cholesterol and sphingolipid in the target liposome membrane. In tissue culture, both SFV fusion and virus exit are strongly cholesterol-dependent when assayed in cholesterol-depleted insect cells. We here describe the preparation of insect cells that while not containing detectable amounts of cholesterol, have adapted to sterol-depleted conditions, resulting in a more permissive phenotype for SFV infection. Although still less efficient at supporting SFV infection than control cholesterol-containing cells, the adapted cells show a 45-fold increase in primary infection by SFV, increased release of progeny virus, and enhanced virus growth kinetics compared to nonadapted cholesterol-depleted cells. The adapted cells are also about 35-fold more permissive for low pH-induced fusion of SFV with the plasma membrane, suggesting that adaptation correlates with a change in the cell membrane.

Original languageEnglish (US)
Pages (from-to)198-205
Number of pages8
JournalVirology
Volume224
Issue number1
DOIs
StatePublished - Oct 1 1996

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Semliki forest virus
Virus Diseases
Cholesterol
Viruses
Cell Membrane
Liposomes
Insects
Alphavirus
Virus Release
Transport Vesicles
Sphingolipids
Membranes
Sterols
Infection Control
Endocytosis
Phenotype
Growth
Infection

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Cholesterol-depleted cells that are relatively permissive for Semliki Forest virus infection. / Marquardt, Marianne T.; Kielian, Margaret.

In: Virology, Vol. 224, No. 1, 01.10.1996, p. 198-205.

Research output: Contribution to journalArticle

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abstract = "Semliki Forest virus (SFV), an enveloped alphavirus, infects cells by endocytosis followed by low pH-triggered fusion of the virus and endocytic vesicle membranes. Progeny virus is released by budding from the cell plasma membrane. In vitro SFV fusion with artificial liposomes is triggered by low pH and is dependent on the presence of cholesterol and sphingolipid in the target liposome membrane. In tissue culture, both SFV fusion and virus exit are strongly cholesterol-dependent when assayed in cholesterol-depleted insect cells. We here describe the preparation of insect cells that while not containing detectable amounts of cholesterol, have adapted to sterol-depleted conditions, resulting in a more permissive phenotype for SFV infection. Although still less efficient at supporting SFV infection than control cholesterol-containing cells, the adapted cells show a 45-fold increase in primary infection by SFV, increased release of progeny virus, and enhanced virus growth kinetics compared to nonadapted cholesterol-depleted cells. The adapted cells are also about 35-fold more permissive for low pH-induced fusion of SFV with the plasma membrane, suggesting that adaptation correlates with a change in the cell membrane.",
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