Cell Type-Specific Targeting with Surface-Engineered Lentiviral Vectors Co-displaying OKT3 Antibody and Fusogenic Molecule
- 223 Downloads
The purpose of this study was to investigate the potential of a T-cell-related targeting method using a lentiviral vector-based gene delivery system.
Materials and Methods
A lentiviral vector system was constructed by co-incorporating an anti-CD3 antibody (OKT3) and a fusogen into individual viral particles. The incorporation of OKT3 and fusogen was analyzed using confocal microscopy and the in vitro transduction efficiency was evaluated using flow cytometry. Blocking reagents (ammonium chloride (NH4Cl) and soluble OKT3 antibody) were added into vector supernatants during transduction to study the mechanism of this two-molecule targeting strategy. To demonstrate the ability of targeted transduction in vivo, Jurkat.CD3 cells were xenografted subcutaneously into the right flank of each mouse and the lentiviral vector was injected subcutaneously on both sides of each mouse 8 h post-injection. Subsequently, the reporter gene (firefly luciferase) expression was monitored using a noninvasive bioluminescence imaging system.
By co-displaying OKT3 and fusogen on the single lentiviral surface, we could achieve targeted delivery of genes to CD3-positive T-cells both in vitro and in vivo.
These results suggest the potential utility of this engineered lentiviral system as a new tool for cell type-directed gene delivery.
KEY WORDSCD3 antigen gene therapy lentiviral vectors targeted gene delivery
We thank April Tai, Lili Yang and Steven Froelich for critical reading of the manuscript, and the USC Norris Center Cell and Tissue Imaging Core. This work was supported by a National Institute of Health grant. The following reagents was obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH: Monoclonal Antibody to HIV-1 p24 (AG3.0) from Dr. Jonathan Allan.
- 2.A. Aiuti, S. Vai, A. Mortellaro, G. Casorati, F. Ficara, G. Andolfi, G. Ferrari, A. Tabucchi, F. Carlucci, H. D. Ochs, L. D. Notarangelo, M. G. Roncarolo, and C. Bordignon. Immune reconstitution in ADA-SCID after PBL gene therapy and discontinuation of enzyme replacement. Nat. Med. 8:423–425 (2002) doi: 10.1038/nm0502-423.PubMedCrossRefGoogle Scholar
- 4.W. R. Drobyski, H. C. Morse 3rd, W. H. Burns, J. T. Casper, and G. Sandford. Protection from lethal murine graft-versus-host disease without compromise of alloengraftment using transgenic donor T cells expressing a thymidine kinase suicide gene. Blood. 97:2506–2513 (2001) doi: 10.1182/blood.V97.8.2506.PubMedCrossRefGoogle Scholar
- 5.M. Maurice, E. Verhoeyen, P. Salmon, D. Trono, S. J. Russell, and F. L. Cosset. Efficient gene transfer into human primary blood lymphocytes by surface-engineered lentiviral vectors that display a T cell-activating polypeptide. Blood. 99:2342–2350 (2002) doi: 10.1182/blood.V99.7.2342.PubMedCrossRefGoogle Scholar
- 6.R. A. Morgan, M. E. Dudley, J. R. Wunderlich, M. S. Hughes, J. C. Yang, R. M. Sherry, R. E. Royal, S. L. Topalian, U. S. Kammula, N. P. Restifo, Z. Zheng, A. Nahvi, C. R. de Vries, L. J. Rogers-Freezer, S. A. Mavroukakis, and S. A. Rosenberg. Cancer regression in patients after transfer of genetically engineered lymphocytes. Science. 314:126–129 (2006) doi: 10.1126/science.1129003.PubMedCrossRefGoogle Scholar
- 7.B. L. Levine, L. M. Humeau, J. Boyer, R. R. MacGregor, T. Rebello, X. Lu, G. K. Binder, V. Slepushkin, F. Lemiale, J. R. Mascola, F. D. Bushman, B. Dropulic, and C. H. June. Gene transfer in humans using a conditionally replicating lentiviral vector. Proc. Natl. Acad. Sci. U.S.A. 103:17372–17377 (2006) doi: 10.1073/pnas.0608138103.PubMedCrossRefGoogle Scholar
- 23.M. Marin, D. Noel, S. Valsesia-Wittman, F. Brockly, M. Etienne-Julan, S. Russell, F. L. Cosset, and M. Piechaczyk. Targeted infection of human cells via major histocompatibility complex class I molecules by Moloney murine leukemia virus-derived viruses displaying single-chain antibody fragment-envelope fusion proteins. J. Virol. 70:2957–2962 (1996).PubMedGoogle Scholar
- 28.P. Roux, P. Jeanteur, and M. Piechaczyk. A versatile and potentially general approach to the targeting of specific cell types by retroviruses: application to the infection of human cells by means of major histocompatibility complex class I and class II antigens by mouse ecotropic murine leukemia virus-derived viruses. Proc. Natl. Acad. Sci. U.S.A. 86:9079–9083 (1989) doi: 10.1073/pnas.86.23.9079.PubMedCrossRefGoogle Scholar
- 31.A. H. Lin, N. Kasahara, W. Wu, R. Stripecke, C. L. Empig, W. F. Anderson, and P. M. Cannon. Receptor-specific targeting mediated by the coexpression of a targeted murine leukemia virus envelope protein and a binding-defective influenza hemagglutinin protein. Hum. Gene Ther. 12:323–332 (2001) doi: 10.1089/10430340150503957.PubMedCrossRefGoogle Scholar
- 32.H. Yang, L. Zeigler, K. I. Joo, T. Cho, Y. Lei, and P. Wang. Gamma-retroviral vectors enveloped with an antibody and an engineered fusogenic protein achieved antigen-specific targeting. Biotechnol. Bioeng. 19:861–872 (2008).Google Scholar
- 37.A. B. Cosimi, R. C. Burton, R. B. Colvin, G. Goldstein, F. L. Delmonico, M. P. LaQuaglia, N. Tolkoff-Rubin, R. H. Rubin, J. T. Herrin, and P. S. Russell. Treatment of acute renal allograft rejection with OKT3 monoclonal antibody. Transplantation. 32:535–539 (1981) doi: 10.1097/00007890-198112000-00018.PubMedCrossRefGoogle Scholar
- 38.A. B. Cosimi, R. B. Colvin, R. C. Burton, R. H. Rubin, G. Goldstein, P. C. Kung, W. P. Hansen, F. L. Delmonico, and P. S. Russell. Use of monoclonal antibodies to T-cell subsets for immunologic monitoring and treatment in recipients of renal allografts. N. Engl. J. Med. 305:308–314 (1981).PubMedGoogle Scholar