Enhancing dengue virus maturation using a stable furin over-expressing cell line

Area of Science:

• Virology
• Molecular Biology
• Cell Biology

Background:

• Flaviviruses require furin-mediated cleavage of the premembrane (prM) protein for maturation.
• Immature flavivirus particles, resulting from inefficient prM cleavage, exhibit distinct antigenic and functional properties.
• Viruses produced in standard tissue culture cells are often less mature than those from primary cells.

Purpose of the Study:

• To develop a cell line for producing more homogenous, mature flavivirus populations.
• To investigate the utility of a furin-expressing cell line for flavivirus production.

Main Methods:

• Generation of a Vero cell line with ectopic expression of human furin (Vero-furin).
• Culturing laboratory-adapted and clinical dengue virus isolates in Vero-furin cells.
• Employing biochemical and structural techniques to analyze viral maturation and prM cleavage.

Main Results:

• Vero-furin cells efficiently support the growth of dengue virus isolates.
• Efficient prM cleavage was observed in flavivirus preparations derived from Vero-furin cells.
• Virions produced in Vero-furin cells showed reduced sensitivity to neutralization by antibodies targeting immature virions.

Conclusions:

• The Vero-furin cell line facilitates the production of homogenous, mature flavivirus particles.
• This engineered cell line is valuable for flavivirus neutralization assays and structural biology studies.
• Enhanced furin expression improves flavivirus maturation, impacting viral properties and immune interactions.