Intrapulmonary Versus Nasal Transduction of Murine Airways With GP64-pseudotyped Viral Vectors

Abstract

Persistent viral vector-mediated transgene expression in the airways requires delivery to cells with progenitor capacity and avoidance of immune responses. Previously, we observed that GP64-pseudotyped feline immunodeficiency virus (FIV)-mediated gene transfer was more efficient in the nasal airways than the large airways of the murine lung. We hypothesized that in vivo gene transfer was limited by immunological and physiological barriers in the murine intrapulmonary airways. Here, we systematically investigate multiple potential barriers to lentiviral gene transfer in the airways of mice. We show that GP64-FIV vector transduced primary cultures of well-differentiated murine nasal epithelia with greater efficiency than primary cultures of murine tracheal epithelia. We further demonstrate that neutrophils, type I interferon (IFN) responses, as well as T and B lymphocytes are not the major factors limiting the transduction of murine conducting airways. In addition, we observed better transduction of GP64-pseudotyped vesicular stomatitis virus (VSV) in the nasal epithelia compared with the intrapulmonary airways in mice. VSVG glycoprotein pseudotyped VSV transduced intrapulmonary epithelia with similar efficiency as nasal epithelia. Our results suggest that the differential transduction efficiency of nasal versus intrapulmonary airways by FIV vector is not a result of immunological barriers or surface area, but rather differential expression of cellular factors specific for FIV vector transduction.

Human, Pig, and Mouse Interferon-Induced Transmembrane Proteins Partially Restrict Pseudotyped Lentiviral Vectors

“…In a previous study, we observed that GP64-FIV transduced mouse nasal airways with greater efficiency than the intrapulmonary airways. 10 This observation was true both in vivo and in welldifferentiated primary cultures derived from mouse nasal septa or trachea. We hypothesized that the differential expression pattern was due to a cellspecific factor.…”

“…We previously demonstrated that FIV pseudotyped with the envelope glycoprotein from baculovirus Autographa californica multinucleocapsid nucleopolyhedrosis virus (GP64) confers apical entry into well-differentiated primary cultures of human, mouse, and pig airway epithelial cells. 4,[8][9][10] We further demonstrated that GP64-FIV efficiently transduced the airways of mice or pigs in vivo, was repeatedly administered without the generation of blocking immune responses, and expressed a reporter gene for the life span of mice. 9,11 Thus, GP64-FIV is a candidate gene therapy reagent for genetic pulmonary diseases such as cystic fibrosis.…”

“…In addition, the transduction efficiency difference was also observed with VSV-G-and Ebola-pseudotyped FIV. 10 These data suggested that a cellular restriction factor was responsible for the difference in transduction efficiency. Here, we investigate whether the IFITM family of proteins contributes to the differential transduction efficiency observed between the nasal and tracheal airways.…”

“…[24][25][26] We previously reported that GP64-FIV transduces mouse nasal airway epithelia with *10-fold greater efficiency than mouse intrapulmonary epithelia. 10 This difference was not the result of surface area, immune responses, or clearance because the difference was recapitulated in well-differentiated primary cultures derived from mouse septa and trachea. In addition, the transduction efficiency difference was also observed with VSV-G-and Ebola-pseudotyped FIV.…”

Lentiviral-mediated phenotypic correction of cystic fibrosis pigs

“…Lentiviral vectors transduce both dividing and nondividing cells, integrate into the host genome, and provide long-term trans-gene expression (10, 11). A feline immunodeficiency virus–based (FIV-based) viral vector pseudotyped with the baculovirus envelope protein GP64 transduces epithelial cells at the apical surface and persistently expresses a transgene of interest in mouse airways (12, 13). We previously demonstrated that FIV pseudotyped with the GP64 envelope from baculovirus efficiently transduces both human and pig airway epithelia (12, 14).…”