Targeting lentiviral vectors to specific cell types in vivo

doi:10.1073/pnas.0604993103

Targeting lentiviral vectors to specific cell types in vivo

*Division of Biology, California Institute of Technology, Pasadena, CA 91125; and
^†Mork Family Department of Chemical Engineering and Materials Science, University of Southern California, Los Angeles, CA 90089

Contributed by David Baltimore, June 15, 2006

Abstract

We have developed an efficient method to target lentivirus-mediated gene transduction to a desired cell type. It involves incorporation of antibody and fusogenic protein as two distinct molecules into the lentiviral surface. The fusogen is constructed by modifying viral envelope proteins, so that they lack the ability to bind to their cognate receptor but still retain the ability to trigger pH-dependent membrane fusion. Thus, the specificity of such a lentiviral vector is solely determined by the antibody, which is chosen to recognize a specific surface antigen of the desired cell type. This specific binding then induces endocytosis of the surface antigen, bringing the lentivirus into an endosome. There, the fusogen responds to the low pH environment and mediates membrane fusion, allowing the virus core to enter the cytosol. Using CD20 as a target antigen for human B cells, we have demonstrated that this targeting strategy is effective both in vitro and in intact animals. This methodology is flexible and can be extended to other forms of cell type-specific recognition to mediate targeting. The only requirement is that the antibody (or other binding protein) must be endocytosed after interaction with its cell surface-binding determinant.

Footnotes

^‡To whom correspondence may be addressed. E-mail: baltimo{at}caltech.edu or pinwang{at}usc.edu
Author contributions: L.Y., D.B., and P.W. designed research; L.Y., L.B., and P.W. performed research; L.Y. and P.W. contributed new reagents/analytic tools; L.Y., L.B., D.B., and P.W. analyzed data; and L.Y., L.A.B., D.B., and P.W. wrote the paper.
Conflict of interest statement: No conflicts declared.
↵ § We have observed that this method can be exploited to target dendritic cells using a membrane-bound monoclonal antibody against the DEC-205 receptor. In addition, we found that incorporation of a membrane-bound form of stem cell factor could target c-kit-positive cells.
Abbreviations:

FM,

fusogenic molecule;

Env,

envelope glycoprotein;

SIN,

Sindbis virus glycoprotein;

αCD20,

anti-human CD20 antibody;

TU,

transduction units;

PBMC,

peripheral blood mononuclear cells;

FPV,

influenza A/fowl plague virus/Rostock/34
Freely available online through the PNAS open access option.