Transduction of Murine Alveolar Macrophages with Adeno-associated Virus 6: Implications for Cell and Gene Therapy

Raw Data for the manuscript "Transduction of Murine Alveolar Macrophages with Adeno-associated Virus 6: Implications for Cell and Gene Therapy".
Abstract:
Alveolar macrophages (AMs) are self-renewing, tissue resident cells of the lung. Their genetic manipulation remains challenging but due to their longevity they are promising tools for gene or cell therapy.
Murine AMs were isolated from C57BL/6J mice and characterized for purity and gene expression profile. AMs were cultured and transduced with different AAV serotypes expressing green fluorescent protein (GFP) to investigate the effects of serotype, multiplicity of infection (MOI), and incubation time, and analyzed by live cell and confocal microscopy. Transducing AMs with AAV6 at a MOI of 87,500 resulted in the highest transduction rate, transgene signal intensity, and expression after 6 days. Activation of AM by lipopolysaccharide (LPS) stimulation significantly increased GFP protein synthesis. AAV transduction did not induce inflammation in AMs and transgene signal intensity was stable after the removal of the LPS stimulus. In conclusion, murine AMs can be successfully cultured in vitro, AAV6 showed the highest transduction efficiency, and activation by LPS significantly increases transgene expression.