Efficient Transduction and Expansion of Ovine Macrophages for Gene Therapy Implementations

Area of Science:

• Veterinary Medicine
• Molecular Biology
• Infectious Diseases

Background:

• Zoonotic bacteria often survive intracellularly within monocyte/macrophage lineages, complicating eradication and causing chronic inflammation.
• Ruminant phagocytes are critical in zoonotic diseases like brucellosis, but their susceptibility to lentiviral vector transduction is poorly understood.
• Lentiviral vectors show promise for targeting intracellular pathogens, but their efficacy in animal models requires investigation.

Purpose of the Study:

• To assess the transduction efficiency of ovine macrophages using lentiviral vectors.
• To evaluate transgene expression and vector copy number (VCN) in ovine macrophages.
• To establish a foundation for gene therapy strategies against intracellular bacterial zoonoses in ruminants.

Main Methods:

• Ovine macrophages were cultured and exposed to a lentiviral vector encoding green fluorescence protein.
• Transduction efficiency was measured by transgene expression and vector copy number (VCN).
• The effect of transduction enhancers, such as polybrene, was evaluated.

Main Results:

• Ovine macrophages demonstrated relative resistance to lentiviral vector transduction, even at high multiplicities of infection.
• The addition of polybrene significantly enhanced transgene expression in transduced ovine macrophages.
• Sustained transgene expression was observed after one week of in vitro culture.

Conclusions:

• Ovine macrophages can be effectively expanded and transduced in culture for potential gene therapy applications.
• Lentiviral vector transduction of ovine macrophages is feasible, albeit requiring optimization with enhancers.
• This study provides a critical benchmark for developing gene therapy targeting zoonotic diseases in ruminant populations.