High-Efficiency Gene Transfer to Primary Monkey Airway Epithelial Cells with Retrovirus Vectors Using the Gibbon Ape Leukemia Virus Receptor

Abstract

The efficiency of retrovirus-mediated gene transfer to primary airway epithelial cells from rhesus monkeys was evaluated. We compared the use of murine amphotropic retrovirus vectors to the use of murine retrovirus vectors containing the envelope (Env) glycoproteins from gibbon ape leukemia virus (GALV). These vectors use distinct receptors to gain entry into host cells. We found that vectors with the GALV Env glycoproteins are up to 10-fold more efficient at transducing genes into primary monkey airway epithelial cells than vectors with the amphotropic Env glycoproteins. Under optimal conditions, up to about 80% of primary monkey airway epithelial cells could be transduced with the vector containing the GALV Env glycoproteins. In addition, we found that delivery of retrovirus vectors to the apical side of polarized airway epithelial cultures was significantly more efficient than delivery to the basal side. These results suggest the feasibility of luminal delivery of retrovirus vectors to the lung.

Influenza M2 envelope protein augments avian influenza hemagglutinin pseudotyping of lentiviral vectors

“…This may reflect an asymmetry in receptor levels for HA-pseudotyped vectors since the difference in transduction between the two sides was greater than that expected from the barrier effect of the collagencoated membrane of the Transwell-col structure. 26 In cultures transduced apically, up to 30% of cells were transduced with the HA-pseudotyped vector as determined by morphometric analysis, whereas basolateral application of virus with the same titer resulted in o5% transduction. In these experiments, the efficiency of FPV HA-mediated apical transduction was found to be similar to VSV-G-mediated basolateral transduction.…”

Effect of Adenoviral Vector Infection on Cell Proliferation in Cultured Primary Human Airway Epithelial Cells

“…The S phase of the cell cycle was measured by incorporation of bromodeoxyuridine (BrdU) into the proUferating cells as previously described (Bayle et al, 1993) with the following modifications. Briefly, cells were pulsed for 2 hr with 4 X 10~* M BrdU (Boehringer Mannheim Corp.) in F12 media.…”

Cell and Gene Therapy for Genetic Diseases: Inherited Disorders Affecting the Lung and Those Mimicking Sudden Infant Death Syndrome

“…Generally, these vectors have been pseudotyped with envelopes that are suitable for targeting receptors present on airway epithelial cells. These have included the vesicular stomatitis virus-G glycoprotein, the Gibbon ape leukemia virus envelope, and an envelope targeting the folate receptor (Bayle et al, 1993;Copreni et al, 2010). In general, the longevity and safety of such systems have been quite good.…”