Efficient lentiviral transduction and transgene expression in primary human B cells

Area of Science:

• Gene Therapy
• Immunology
• Virology

Background:

• Primary human B cells are a key target for gene therapy but show resistance to lentiviral vector transduction.
• Low transgene expression further limits the utility of lentiviral vectors (LVVs) in B cells.

Purpose of the Study:

• To investigate the transduction efficiency of gibbon-ape leukemia virus (GALV) Env-pseudotyped LVVs in primary human B cells.
• To develop an optimized LVV for enhanced transgene expression in B cells for therapeutic applications.

Main Methods:

• Optimized transduction kinetics and multiplicities of infection were established for GALV-pseudotyped LVVs.
• A novel LVV was engineered by combining enhancer (Eμ) and matrix/scaffold-attachment regions (MARs) with the spleen focus-forming virus (SFFV) promoter.
• Transduction efficiencies and transgene expression levels were assessed in primary human B cells and B cell lines.

Main Results:

• Over 50% transduction efficiency was achieved in CD40L-activated primary human B cells using GALV-pseudotyped LVVs.
• GALV-pseudotyped LVVs yielded over 10-fold more transduced cells compared to measles virus glycoprotein-pseudotyped LVVs.
• The novel, optimized LVV demonstrated a mean 3-fold increase in transgene expression in B cells.

Conclusions:

• An efficient protocol for lentiviral transduction of primary human B cells was established using GALV Env-pseudotyped LVVs.
• Vector modification significantly improved transgene expression in B cells, enhancing their suitability for gene therapy.
• The findings pave the way for improved gene-therapeutic strategies targeting human B cells.