title: Adenoviral targeting using genetically incorporated camelid single variable domains.
abstract: The unique ability of human adenovirus serotype 5 (Ad5) to accomplish efficient transduction has allowed the use of Ad5-based vectors for a range of gene therapy applications. Several strategies have been developed to alter tropism of Ad vectors to achieve a cell-specific gene delivery by using fiber modifications via genetic incorporation of targeting motifs. In this study, we have explored the utility of novel anti-human carcinoembryonic antigen (hCEA) single variable domains derived from heavy chain (VHH) camelid family of antibodies to achieve targeted gene transfer. To obtain anti-CEA VHHs, we produced a VHH-display library from peripheral blood lymphocytes RNA of alpacas at the peak of immune response to the hCEA antigen (Ag). We genetically incorporated an anti-hCEA VHH into a de-knobbed Ad5 fiber-fibritin chimera and demonstrated selective targeting to the cognate epitope expressed on the membrane surface of target cells. We report that the anti-hCEA VHH used in this study retains Ag recognition functionality and provides specificity for gene transfer of capsid-modified Ad5 vectors. These studies clearly demonstrated the feasibility of retargeting of Ad5-based gene transfer using VHHs.
authors.affiliation: Department of Radiation Oncology, School of Medicine, Washington University in St Louis, St Louis, MO, USA.
authors.forename: Sergey A
authors.initials: SA
authors.lastname: Kaliberov
authors.affiliation: Department of Radiation Oncology, School of Medicine, Washington University in St Louis, St Louis, MO, USA.
authors.forename: Lyudmila N
authors.initials: LN
authors.lastname: Kaliberova
authors.affiliation: Department of Radiation Oncology, School of Medicine, Washington University in St Louis, St Louis, MO, USA.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Buggio
authors.affiliation: Department of Infectious Disease and Global Health, Tufts Cummings School of Veterinary Medicine, North Grafton, MA, USA.
authors.forename: Jacqueline M
authors.initials: JM
authors.lastname: Tremblay
authors.affiliation: Department of Infectious Disease and Global Health, Tufts Cummings School of Veterinary Medicine, North Grafton, MA, USA.
authors.forename: Charles B
authors.initials: CB
authors.lastname: Shoemaker
authors.affiliation: Department of Radiation Oncology, School of Medicine, Washington University in St Louis, St Louis, MO, USA.
authors.forename: David T
authors.initials: DT
authors.lastname: Curiel
mesh_terms: D000256:Adenoviridae / Q000502:physiology*; D000818:Animals; D000918:Antibody Specificity; D002161:Camelids, New World; D036022:Capsid Proteins / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D002272:Carcinoembryonic Antigen / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D045744:Cell Line, Tumor; D005240:Feasibility Studies; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage / Q000502:physiology*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008297:Male; D010446:Peptide Fragments / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism; D014161:Transduction, Genetic; D014764:Viral Proteins / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D056189:Viral Tropism; D014771:Virion / Q000502:physiology
chemical_list: D036022:Capsid Proteins; D002272:Carcinoembryonic Antigen; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; C057839:fibritin protein, Enterobacteria phage T4