PEGylation of a vesicular stomatitis virus G pseudotyped lentivirus vector prevents inactivation in serum.
One disadvantage of vesicular stomatitis virus G (VSV-G) pseudotyped lentivirus vectors for clinical application is inactivation of the vector by human serum complement. To prevent this, monomethoxypoly(ethylene) glycol was conjugated to a VSV-G-human immunodeficiency virus vector expressing Escherichia coli beta-galactosidase. The modification did not affect transduction efficiency in vitro and protected the vector from inactivation in complement-active human and mouse sera. Blood from mice dosed intravenously with either the unmodified or the PEGylated virus particles was assayed for active vector by a limiting-dilution assay to evaluate transduction efficiency and for p24, an indicator of the total number of virus particles present. PEGylation extended the circulation half-life of active vector by a factor of 5 and reduced the rate of vector inactivation in the serum by a factor of 1,000. Pharmacokinetic profiles for the total number of virus particles present in the circulation were unaffected by PEGylation. Modification of the vector with poly(ethylene) glycol significantly enhanced transduction efficiency in the bone marrow and in the spleen 14 days after systemic administration of the virus. These results, in concert with the pharmacokinetic profiles, indicate that PEGylation does protect the virus from inactivation in the serum and, as a result, improves the transduction efficiency of VSV-G pseudotyped lentivirus vectors in susceptible organs in vivo.
Croyle
Maria A
MA
Division of Pharmaceutics, College of Pharmacy, The University of Texas at Austin Austin, Texas 78712, USA. macroyle@mail.utexas.edu
Callahan
Shellie M
SM
Auricchio
Alberto
A
Schumer
Gregg
G
Linse
Klause D
KD
Wilson
James M
JM
Brunner
Lane J
LJ
Kobinger
Gary P
GP
D000818:Animals; D001853:Bone Marrow / Q000821:virology; D019075:Electrophoresis, Capillary; D005822:Genetic Vectors / Q000008:administration & dosage / Q000737:chemistry*; D015497:HIV-1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009500:Neutralization Tests; D011092:Polyethylene Glycols / Q000737:chemistry*; D013154:Spleen / Q000821:virology; D014161:Transduction, Genetic; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics* / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
C028391:G protein, vesicular stomatitis virus; D007106:Immune Sera; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D011092:Polyethylene Glycols