High-throughput screening of viral entry inhibitors using pseudotyped virus

Area of Science:

• Virology
• Infectious Diseases
• Drug Discovery

Background:

• Virus entry is a key therapeutic target to prevent infection and limit drug resistance.
• Studying BSL-4 viruses like Ebola requires specialized, high-containment laboratories.
• Developing antivirals against BSL-4 pathogens is challenging due to safety and containment requirements.

Purpose of the Study:

• To present a method for preparing and utilizing pseudotyped viruses for antiviral drug screening.
• To enable high-throughput screening (HTS) of viral entry inhibitors in BSL-2 facilities.
• To facilitate the discovery of therapeutics against BSL-4 viruses.

Main Methods:

• Generation of pseudotyped viruses by replacing the envelope glycoprotein of a BSL-2 virus with that of a BSL-4 virus.
• Utilizing lentiviral vectors for the production of pseudotyped viral particles.
• Employing pseudotyped viruses in high-throughput screening assays to identify entry inhibitors.

Main Results:

• Pseudotyped viruses accurately mimic the entry pathway of the original BSL-4 virus.
• These surrogate viruses are replication-incompetent, allowing safe handling in BSL-2 laboratories.
• The method supports efficient screening of numerous compounds for antiviral activity.

Conclusions:

• Pseudotyped viruses are a valuable tool for safe and effective antiviral drug discovery against high-risk pathogens.
• This approach overcomes the limitations of BSL-4 containment for early-stage antiviral research.
• The protocols described facilitate the development of novel therapeutics targeting viral entry mechanisms.