Abstract
Purpose
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.
Results
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.
Conclusions
These results suggest the potential utility of this engineered lentiviral system as a new tool for cell type-directed gene delivery.
Similar content being viewed by others
References
Y. Chernajovsky, G. Adams, K. Triantaphyllopoulos, M. F. Ledda, and O. L. Podhajcer. Pathogenic lymphoid cells engineered to express TGF beta 1 ameliorate disease in a collagen-induced arthritis model. Gene. Ther. 4:553–559 (1997) doi:10.1038/sj.gt.3300436.
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.
E. Verhoeyen, V. Dardalhon, O. Ducrey-Rundquist, D. Trono, N. Taylor, and F. L. Cosset. IL-7 surface-engineered lentiviral vectors promote survival and efficient gene transfer in resting primary T lymphocytes. Blood. 101:2167–2174 (2003) doi:10.1182/blood-2002-07-2224.
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.
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.
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.
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.
M. Sadelain, I. Riviere, and R. Brentjens. Targeting tumours with genetically enhanced T lymphocytes. Nat. Rev. Cancer. 3:35–45 (2003) doi:10.1038/nrc971.
M. T. Stephan, V. Ponomarev, R. J. Brentjens, A. H. Chang, K. V. Dobrenkov, G. Heller, and M. Sadelain. T cell-encoded CD80 and 4–1BBL induce auto- and transcostimulation, resulting in potent tumor rejection. Nat. Med. 13:1440–1449 (2007) doi:10.1038/nm1676.
T. N. Schumacher. T-cell-receptor gene therapy. Nat. Rev. Immunol. 2:512–519 (2002) doi:10.1038/nri841.
O. J. Muller, F. Kaul, M. D. Weitzman, R. Pasqualini, W. Arap, J. A. Kleinschmidt, and M. Trepel. Random peptide libraries displayed on adeno-associated virus to select for targeted gene therapy vectors. Nat. Biotechnol. 21:1040–1046 (2003) doi:10.1038/nbt856.
D. G. Miller, M. A. Adam, and A. D. Miller. Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection. Mol. Cell Biol. 10:4239–4242 (1990).
L. Naldini, U. Blomer, P. Gallay, D. Ory, R. Mulligan, F. H. Gage, I. M. Verma, and D. Trono. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science. 272:263–267 (1996) doi:10.1126/science.272.5259.263.
E. Costello, M. Munoz, E. Buetti, P. R. Meylan, H. Diggelmann, and M. Thali. Gene transfer into stimulated and unstimulated T lymphocytes by HIV-1-derived lentiviral vectors. Gene Ther. 7:596–604 (2000) doi:10.1038/sj.gt.3301135.
D. B. Kohn. Lentiviral vectors ready for prime-time. Nat. Biotechnol. 25:65–66 (2007) doi:10.1038/nbt0107-65.
L. E. Ailles, and L. Naldini. HIV-1-derived lentiviral vectors. Curr. Top. Microbiol. Immunol. 261:31–52 (2002).
J. Cronin, X. Y. Zhang, and J. Reiser. Altering the tropism of lentiviral vectors through pseudotyping. Curr. Gene Ther. 5:387–398 (2005) doi:10.2174/1566523054546224.
V. Sandrin, S. J. Russell, and F. L. Cosset. Targeting retroviral and lentiviral vectors. Curr. Top. Microbiol. Immunol. 281:137–178 (2003).
E. Verhoeyen, and F. L. Cosset. Surface-engineering of lentiviral vectors. J. Gene Med. 6(Suppl 1):S83–S94 (2004) doi:10.1002/jgm.494.
R. Waehler, S. J. Russell, and D. T. Curiel. Engineering targeted viral vectors for gene therapy. Nat. Rev. Genet. 8:573–587 (2007) doi:10.1038/nrg2141.
D. Lavillette, S. J. Russell, and F. L. Cosset. Retargeting gene delivery using surface-engineered retroviral vector particles. Curr. Opin. Biotechnol. 12:461–466 (2001) doi:10.1016/S0958-1669(00)00246-9.
S. Ager, B. H. Nilson, F. J. Morling, K. W. Peng, F. L. Cosset, and S. J. Russell. Retroviral display of antibody fragments; interdomain spacing strongly influences vector infectivity. Hum. Gene Ther. 7:2157–2164 (1996) doi:10.1089/hum.1996.7.17-2157.
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).
S. Chowdhury, K. A. Chester, J. Bridgewater, M. K. Collins, and F. Martin. Efficient retroviral vector targeting of carcinoembryonic antigen-positive tumors. Mol. Ther. 9:85–92 (2004) doi:10.1016/j.ymthe.2003.10.004.
S. Funke, A. Maisner, M. D. Muhlebach, U. Koehl, M. Grez, R. Cattaneo, K. Cichutek, and C. J. Buchholz. Targeted cell entry of lentiviral vectors. Mol. Ther. 16:1427–1436 (2008) doi:10.1038/mt.2008.128.
K. Morizono, G. Bristol, Y. M. Xie, S. K. Kung, and I. S. Chen. Antibody-directed targeting of retroviral vectors via cell surface antigens. J. Virol. 75:8016–8020 (2001) doi:10.1128/JVI.75.17.8016-8020.2001.
K. Morizono, Y. Xie, G. E. Ringpis, M. Johnson, H. Nassanian, B. Lee, L. Wu, and I. S. Chen. Lentiviral vector retargeting to P-glycoprotein on metastatic melanoma through intravenous injection. Nat. Med. 11:346–352 (2005) doi:10.1038/nm1192.
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.
A. L. Boerger, S. Snitkovsky, and J. A. Young. Retroviral vectors preloaded with a viral receptor-ligand bridge protein are targeted to specific cell types. Proc. Natl. Acad. Sci. U.S.A. 96:9867–9872 (1999) doi:10.1073/pnas.96.17.9867.
L. Yang, L. Bailey, D. Baltimore, and P. Wang. Targeting lentiviral vectors to specific cell types in vivo. Proc. Natl. Acad. Sci. U.S.A. 103:11479–11484 (2006) doi:10.1073/pnas.0604993103.
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.
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).
C. Lois, E. J. Hong, S. Pease, E. J. Brown, and D. Baltimore. Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science. 295:868–872 (2002) doi:10.1126/science.1067081.
A. L. Szymczak, C. J. Workman, Y. Wang, K. M. Vignali, S. Dilioglou, E. F. Vanin, and D. A. Vignali. Correction of multi-gene deficiency in vivo using a single ‘self-cleaving’ 2A peptide-based retroviral vector. Nat. Biotechnol. 22:589–594 (2004) doi:10.1038/nbt957.
J. Fang, J.-J. Qian, S. Yi, T. C. Harding, G. H. Tu, M. VanRoey, and K. Jooss. Stable antibody expression at therapeutic levels using the 2A peptide. Nat. Biotech. 23:584–590 (2005) doi:10.1038/nbt1087.
K. I. Joo, and P. Wang. Visualization of targeted transduction by engineered lentiviral vectors. Gene Ther. 15:1348–1396 (2008) doi:10.1038/gt.2008.87.
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.
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).
M. G. Rudolph, R. L. Stanfield, and I. A. Wilson. How TCRs bind MHCs, peptides, and coreceptors. Annu. Rev. Immunol. 24:419–466 (2006) doi:10.1146/annurev.immunol.23.021704.115658.
M. Reth. Antigen receptors on B lymphocytes. Annu. Rev. Immunol. 10:97–121 (1992) doi:10.1146/annurev.iy.10.040192.000525.
Y. E. Lu, T. Cassese, and M. Kielian. The cholesterol requirement for sindbis virus entry and exit and characterization of a spike protein region involved in cholesterol dependence. J. Virol. 73:4272–4278 (1999).
I. Mellman, R. Fuchs, and A. Helenius. Acidification of the endocytic and exocytic pathways. Annu. Rev. Biochem. 55:663–700 (1986) doi:10.1146/annurev.bi.55.070186.003311.
M. Kielian, and F. A. Rey. Virus membrane-fusion proteins: more than one way to make a hairpin. Nat. Rev. Microbiol. 4:67–76 (2006) doi:10.1038/nrmicro1326.
D. L. Gibbons, M. C. Vaney, A. Roussel, A. Vigouroux, B. Reilly, J. Lepault, M. Kielian, and F. A. Rey. Conformational change and protein–protein interactions of the fusion protein of Semliki Forest virus. Nature. 427:320–325 (2004) doi:10.1038/nature02239.
M. Umashankar, C. Sanchez-San Martin, M. Liao, B. Reilly, A. Guo, G. Taylor, and M. Kielian. Differential cholesterol binding by class II fusion proteins determines membrane fusion properties. J. Virol. 82:9245–9253 (2008) doi:10.1128/JVI.00975-08.
Acknowledgements
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.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yang, H., Joo, KI., Ziegler, L. et al. Cell Type-Specific Targeting with Surface-Engineered Lentiviral Vectors Co-displaying OKT3 Antibody and Fusogenic Molecule. Pharm Res 26, 1432–1445 (2009). https://doi.org/10.1007/s11095-009-9853-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11095-009-9853-y