TY - JOUR
T1 - Adenoviral targeting using genetically incorporated camelid single variable domains
AU - Kaliberov, Sergey A.
AU - Kaliberova, Lyudmila N.
AU - Buggio, Maurizio
AU - Tremblay, Jacqueline M.
AU - Shoemaker, Charles B.
AU - Curiel, David T.
N1 - Funding Information:
We thank Cynthia L Marich from the Biologic Therapeutics Center, Department of Radiation Oncology, School of Medicine, Washington University in St Louis for her assistance in preparing the manuscript. The authors are grateful to Jesse J Parry from the Department of Radiation Oncology, School of Medicine, Washington University in St Louis for his assistance with FACS. The authors are grateful to Dr Daniela Bedenice, Dr Jean Mukherjee, Karen Baldwin, Kwasi Ofori, and Courtney Boucher from the Tufts Cummings School of Veterinary Medicine for their assistance with alpaca immunization, bleeding and PBL preparation. This work was supported in part by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, under award number U54 AI057159 and R21/R33 AI101403 (CBS), as well as 5P50 CA101955 and 5R01 CA154697 (DTC). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Allergy and Infectious Diseases or the National Institutes of Health.
PY - 2014/8
Y1 - 2014/8
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84905174041&partnerID=8YFLogxK
U2 - 10.1038/labinvest.2014.82
DO - 10.1038/labinvest.2014.82
M3 - Article
C2 - 24933423
AN - SCOPUS:84905174041
SN - 0023-6837
VL - 94
SP - 893
EP - 905
JO - Laboratory Investigation
JF - Laboratory Investigation
IS - 8
ER -