Gene Transfer to Respiratory Epithelia with Lentivirus Pseudotyped with Jaagsiekte Sheep Retrovirus Envelope Glycoprotein

Abstract

A feline immunodeficiency virus (FIV)-based lentiviral vector was pseudotyped to identify envelope (env) glycoproteins that direct efficient gene transfer to pulmonary epithelia for the treatment or prevention of lung diseases. The envelope glycoprotein from the Jaagsiekte sheep retrovirus (JSRV) is a candidate under investigation. We utilized high titer FIV vector (>10(8) TU/ml) pseudotyped with the JSRV env glycoprotein (JSRVFIV) to study the transduction of polarized primary cultures of human airway epithelia and receptor/vector interactions. The reported receptor for JSRV, hyaluronidase 2 (HYAL2), is a GPI-linked protein. We expressed FLAG-tagged HYAL2 in polarized airway epithelia using an adenoviral vector and documented that the HYAL2 protein sorts predominantly to the apical surface. Of interest, the efficiency of gene transfer with apically applied JSRV-FIV was markedly less than FIV pseudotyped with VSV-G, even in Ad-HYAL2 complemented epithelia. The inefficient gene transfer with JSRV-FIV in HYAL2 complemented cells suggests that factors other than receptor abundance limit apical gene transfer efficiency with this envelope. JSRV-FIV transduced the distal lung epithelia of rabbits in vivo and transduced primary cultures of rabbit type II cells with 100-fold greater efficiency than primary cultures of rabbit tracheal cells. These data indicate that a lentivirus pseudotyped with the JSRV envelope glycoprotein transduces type II cells with greater efficiency than conducting airway epithelia and provides an example of glycoprotein-mediated cell-specific tropism within a tissue with a widely heterogeneous cell population.

Reactive Oxygen Species and Hyaluronidase 2 Regulate Airway Epithelial Hyaluronan Fragmentation

“…Here we confirmed that, as previously reported by us and others (26,30,46), GPI-anchored and soluble forms of Hyal2 are restricted to the apical compartment of airway epithelium, suggesting a specific luminal function, unrelated to the turnover of HA in the bronchial subepithelial tissue.…”

“…Forms Are Present Apically in NHBE Cells-We and others (26,46) have previously reported that Hyal2 is localized at the apical membrane of the tracheal epithelium. Here we confirmed that Hyal2 is associated with the cell membrane of NHBE cells through a GPI anchor, and limited to the luminal surface (Fig.…”

Optimization of Feline Immunodeficiency Virus Vectors for RNA Interference

“…Lentiviral vectors are well suited for gene transfer because they provide long-term, heritable gene expression by integrating into host cell chromosomes (28). In addition, they can be modified to transduce numerous cell types, including postmitotic neurons and difficult-to-transduce primary neural progenitor cells (NPCs) (9,15,18,23,28,37,38,42). The most well developed lentiviral vector systems are based on human immunodeficiency virus type 1 (HIV-1), and several publications have described delivery of inhibitory RNAs using HIV-1-based lentiviral vectors in vitro and in vivo (20, 22, 26, 32-34, 45, 48).…”

Lentiviral transduction of the murine lung provides efficient pseudotype and developmental stage-dependent cell-specific transgene expression

“…14,18 Recent interest has converged on pseudotyping; be it of adeno-associated virus (AAV)-2 genomes with novel AAV serotypes 21,36 or of integrating lentiviral vectors with viral envelopes from lung pathogens. 28,29,[31][32][33]37 The Ebola virus envelope glycoprotein has been successfully used to pseudotype lentivirus and has achieved efficient transduction of the murine lung epithelium and human explants 33,34 although generation of consistently high viral titres has been problematic. Recently the baculovirus gp64 envelope has been shown to produce significant expression in the nasal epithelia of adult mice up to 1 year after application.…”