Antibody-Mediated Targeting of Replication-Competent Retroviral Vectors
| Autor: | Jinha M. Park, Chien Kuo Tai, Noriyuki Kasahara, Christopher R. Logg, W. French Anderson, Michael F. Press |
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| Rok vydání: | 2003 |
| Předmět: |
Receptor
ErbB-2 Recombinant Fusion Proteins viruses Genetic Vectors Green Fluorescent Proteins Breast Neoplasms Biology Virus Replication Cell Line Mice Retrovirus Viral Envelope Proteins Viral envelope Tandem repeat Transduction Genetic Cell Line Tumor Murine leukemia virus Genetics Animals Humans Vector (molecular biology) Staphylococcal Protein A Molecular Biology Tropism Virion Antibodies Monoclonal Flow Cytometry biology.organism_classification Virology Protein Structure Tertiary Leukemia Virus Murine Kinetics Luminescent Proteins DNA Viral Monoclonal NIH 3T3 Cells Pseudotyping Molecular Medicine Female Plasmids |
| Zdroj: | Human Gene Therapy. 14:789-802 |
| ISSN: | 1557-7422 1043-0342 |
| DOI: | 10.1089/104303403765255174 |
| Popis: | Replication-competent murine leukemia virus (MLV) vectors can be engineered to achieve high efficiency gene transfer to solid tumors in vivo and tumor-restricted replication, however their safety can be further enhanced by redirecting tropism of the virus envelope. We have therefore tested the targeting capability and replicative stability of ecotropic and amphotropic replication-competent retrovirus (RCR) vectors containing two tandem repeats from the immunoglobulin G-binding domain of Staphylococcal protein A inserted into the proline-rich "hinge" region of the envelope, which enables modular use of antibodies of various specificities for vector targeting. The modified envelopes were efficiently expressed and incorporated into virions, were capable of capturing monoclonal anti-HER2 antibodies, and mediated efficient binding of the virus-antibody complex to HER2-positive target cells. While infectivity was markedly reduced by pseudotyping with targeted envelopes alone, coexpression of wild-type envelope rescued efficient cellular entry. Both ecotropic and amphotropic RCR vector/anti-HER2 antibody complexes achieved significant enhancement of transduction on murine target cells overexpressing HER2, which could be competed by preincubation with excess free antibodies. Interestingly, HER2-expressing human breast cancer cells did not show enhancement of transduction despite efficient antibody-mediated cell surface binding, suggesting that target cell-specific parameters markedly affect the efficiency of post-binding entry processes. Serial replication of targeted vectors resulted in selection of Z domain deletion variants, but reduction of the overall size of the vector genome enhanced its stability. Application of antibody-mediated targeting to the initial localization of replication-competent virus vectors to tumor sites will thus require optimized target selection and vector design. |
| Databáze: | OpenAIRE |
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