Lentiviral Vectors Pseudotyped with Envelope Glycoproteins Derived from Gibbon Ape Leukemia Virus and Murine Leukemia Virus 10A1

Abstract

Lentiviral vectors pseudotyped with the envelope glycoproteins (Env) of amphotropic murine leukemia virus (MLV) and the G protein of vesicular stomatitis virus (VSV-G) have been successfully used in recent preclinical gene therapy studies. We report here the generation of infectious HIV-1-derived vector particles pseudotyped with the Env of the molecular clone 10A1 of MLV and with chimeric envelope glycoprotein variants derived from gibbon ape leukemia virus (GaLV) and MLV. Formation of infectious HIV-1 (GaLV) pseudotype vectors was only possible with the substitution of the cytoplasmic tail of GaLV Env with that of MLV. The lentiviral vectors exhibited a host cell range identical with that of MLV(GaLV) and MLV(10A1) vectors, which are known to enter cells either via the GaLV-receptor Glvr-1 (Pit-1) or via the amphotropic receptor Ram-1 (Pit-2) in addition to Glvr-1, respectively. Thus, HIV-1(GaLV) and HIV-1(10A1) pseudotype vectors may be useful for efficient gene transfer into a variety of human tissues like primary human hematopoietic cells.

Suspension packaging cell lines for the simplified generation of T-cell receptor encoding retrovirus vector particles

“…20,21 The retroviral vector pLEGSN encodes the enhanced GFP gene and the neomycin resistance gene. The retroviral vector MP71-tcr26 encodes a human TCR (Va20-Ja22 [TRAV4*01-TRAJ22*01] and Vb22-Jb2.7 [TRBV2*03-TRBJ2-7*01]) isolated from a tumor-infiltrating T-cell clone (TIL-26) of a renal cell carcinoma patient.…”

Surface-engineering of lentiviral vectors

“…For example, onco-retrovirus vectors pseudotyped with the envelope of the gibbon ape leukemia virus [32,33] or the cat endogenous retrovirus envelope (RD114) allow efficient transduction of HSCs [34]. However, incorporation of these envelope glycoproteins in HIV particles is very inefficient [22,[35][36][37] and requires modifications of their cytoplasmic tails.…”

“…There are specific examples of restriction of pseudotype formation, notably between glycoproteins and viral cores derived from different retrovirus families such as GALV and RD114 lentiviral pseudotypes [22,35,37,[47][48][49][50]. Actually, functional associations between viral glycoproteins and viral cores are unpredictable, in large part because of our insufficient knowledge of the mechanisms that dictate assembly of retroviral particles.…”

“…This suggested, rather, a defect at the level of glycoprotein incorporation on the lentiviral cores. Indeed, the glycoproteins of type C and D mammalian retroviruses, like the GALV and the RD114 viruses, have been shown to harbour in their cytoplasmic tail a determinant that restricts incorporation on lentiviral cores [22,35,37]. The relatively short cytoplasmic tails of type C/D mammalian retrovirus Env, of about 30-40 amino acids length, harbour a 15-20 amino-acid-long carboxy-terminal peptide -named R for MLVs -whose cleavage by the homologous viral core protease is required to activate the fusion potential of the glycoprotein [58][59][60].…”

“…Indeed, replacement of the cytoplasmic tail of RD114 and GALV glycoproteins with that of MLV-A glycoprotein resulted in strongly increased incorporation of either glycoprotein on lentiviral cores for the RD114 glycoprotein and for the GALV glycoprotein [22,35,37]. These chimeric GALV and RD114 glycoproteins (named GALV/TR and RD114/TR) preserved the host range of the initial glycoproteins and conferred 25-fold increased titres compared with SIV vectors pseudotyped with the wt glycoproteins.…”