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Published on: October 30, 2018
Protocol for generating human CAR-engineered macrophages by Vpx-containing lentivirus.
Yun Gao1, Xiaobin Fang1, Luo Zhang2
1RocRock Biotechnology (Suzhou), Suzhou 215000, China.
STAR Protocols
|September 29, 2024
Summary
Chimeric antigen receptor macrophage (CAR-M) therapy is advanced by a new protocol. This method uses the viral protein Vpx to enhance lentivirus infection of human macrophages, overcoming a key barrier.
Area of Science:
- Immunology
- Virology
- Cell Biology
Background:
- Human macrophages are difficult to infect with lentiviruses, hindering CAR-M therapy development.
- HIV-1-based lentiviral vectors are commonly used but show low efficiency in human macrophages.
Purpose of the Study:
- To present a protocol for generating CAR-engineered macrophages with enhanced lentiviral transduction efficiency.
- To overcome the limitation of low lentiviral infectivity in human macrophages for CAR-M therapy.
Main Methods:
- Incorporation of viral protein Vpx (from SIV/HIV-2) into lentivirus vectors.
- Detailed steps for cell cultivation, lentivirus production, concentration, and infection.
- Assessment of CAR-engineered macrophage generation efficiency.
Main Results:
- The Vpx-containing lentivirus vector significantly enhances infection efficiency in human macrophages.
- The protocol facilitates the generation of chimeric antigen receptor (CAR)-engineered macrophages.
Conclusions:
- This protocol provides a foundation for studying macrophage engineering, particularly for CAR-M therapy.
- Vpx incorporation is a viable strategy to improve lentiviral transduction of human macrophages.
