Targeted adenovirus-mediated gene delivery to T cells via CD3.
T cells are primary targets in numerous gene therapy protocols. However, the use of subgroup C adenovirus serotype 2 or 5 (Ad2 or Ad5) as a vector to transduce T cells is limited by its poor transduction efficiency for these cells. In this report we show that poor T-cell transduction results from these cells lacking both the primary Ad2-Ad5 receptor, used in attachment, and the secondary Ad receptor, which mediates entry of most adenovirus serotypes. These deficiencies were overcome by using a bispecific antibody (bsAb) with specificities for human CD3 and for a FLAG epitope genetically introduced into Ad5 (Ad.FLAG) to redirect the virus to human T cells. The anti-FLAG x anti-CD3 bsAb increased Ad.FLAG binding 30-fold, induced the efficient uptake of Ad.FLAG into the cells, and led to a 100- to 500-fold increase in the transduction of resting T cells. Moreover, fluorescence-activated cell sorter analysis showed that 25 to 90% of the T cells were transduced by the bsAb-complexed Ad.FLAG at multiplicities of infection between 20 and 100 active particles per cell. These results demonstrate that bsAbs can target Ad to non-Ad receptors on cells that are normally resistant to Ad, resulting in their efficient and specific transduction.
Wickham
T J
TJ
GenVec, Inc., Rockville, Maryland 20852-1709, USA. wickham@genvec.com
Lee
G M
GM
Titus
J A
JA
Sconocchia
G
G
Bakács
T
T
Kovesdi
I
I
Segal
D M
DM
D000260:Adenoviruses, Human / Q000235:genetics / Q000502:physiology; D018033:Antibodies, Bispecific; D017252:CD3 Complex / Q000276:immunology / Q000502:physiology*; D003587:Cytomegalovirus; D005434:Flow Cytometry; D017930:Genes, Reporter; D005822:Genetic Vectors; D005966:Glucuronidase / Q000096:biosynthesis; D006801:Humans; D019169:Jurkat Cells; D008213:Lymphocyte Activation; D009842:Oligopeptides; D010455:Peptides; D011401:Promoter Regions, Genetic; D011991:Receptors, Virus / Q000276:immunology / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis; D011995:Recombination, Genetic; D013601:T-Lymphocytes / Q000276:immunology / Q000502:physiology*; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000096:biosynthesis
D018033:Antibodies, Bispecific; D017252:CD3 Complex; D009842:Oligopeptides; D010455:Peptides; D011991:Receptors, Virus; D011994:Recombinant Proteins; C085706:FLAG peptide; D001616:beta-Galactosidase; D005966:Glucuronidase