Abstract
We examined the ability of recombinant adeno-associated virus (rAAV) to transfer regulated gene expression into T cell lines. An AAV-based vector containing the neomycin resistance gene and expressing the firefly luciferase (luc) gene under the regulatory control of the interleukin 2 promoter (pAAV-luc) was generated and adenovirus-free rAAV (rAAV-luc) was produced from this vector. Transfection of pAAV-luc into the human T cell line Jurkat resulted in luciferase expression while infection of Jurkat T cells with rAAV-luc resulted in significant luciferase expression only after selection for neomycin-resistant cells. Long-term growth of transduced Jurkat T cells showed that there was no detectable constitutive expression of luciferase and that luciferase gene expression remained inducible for at least 180 days. Luciferase expression was activated by PMA and ionomycin and by anti-CD3 antibodies and was inhibited by cyclosporin A. Examination of G418-resistant clones showed that rAAV-luc had integrated into the host chromosomes but that some of the clones lost some of the transferred DNA or lost expression from the transferred DNA. These results indicate that rAAV can transfer and integrate regulated gene expression into T cell lines but that the transferred genetic material may be lost or its expression may be silenced over time.
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References
Berns K, Linden R . The cryptic life style of adeno-associated virus BioEssays 1994 17: 237–245
Linden R et al. Site-specific integration by adeno-associated virus Proc Natl Acad Sci USA 1996 93: 11288–11294
Kotin R . Prospects for the use of adeno-associated virus as a vector for human gene therapy Hum Gene Ther 1994 5: 793–801
Xiao X, Xiao W, Li J, Samulski J . A novel 165-base-pair terminal repeat sequence is the sole cis requirement for the adeno-associated virus life cycle J Virol 1997 71: 941–948
Miller J et al. Recombinant adeno-assocoated virus (rAAV)-mediated expression of a human r-globin gene in human progenitor-derived erythroid cells Proc Natl Acad Sci USA 1994 91: 10183–10187
Zhou SZ et al. Adeno-associated virus 2-mediated gene transfer in murine hematopoietic progenitor cells Exp Hematol 1993 21: 928–933
Zhou SZ et al. Adeno-associated virus 2-mediated high efficiency gene transfer into immature and mature subsets of hematopoietic progenitor cells in human umbilical cord blood J Exp Med 1994 179: 1867–1875
Podsakoff G, Wong KKJ, Chatterjee S . Efficient gene transfer into nondividing cells by adeno-associated virus-based vectors J Virol 1994 68: 5656–5666
Goodman S et al. Recombinant adeno-associated virus mediated gene transfer into hematopoietic progenitor cells Blood 1994 84: 1492–1500
Walsh CE et al. Phenotypic correction of Fanconi Anemia in human hematopoietic cells with a recombinant adeno-associated virus vector J Clin Invest 1994 94: 1440–1448
Einerhand M et al. Regulated high-level human B-globin gene expression in erythroid cells following recombinant adeno-associated virus-mediated gene transfer Gene Therapy 1995 2: 336–343
Chiorini J et al. High-efficiency transfer of the T cell co-stimulatory molecule B7–2 to lymphoid cells using high-titer recombinant adeno-associated virus vectors Hum Gene Ther 1995 6: 1531–1541
Samulski RJ, Chang L-S, Shenk T . Helper-free stocks of recombinant adeno-associated viruses: normal integration does not require viral gene expression J Virol 1989 63: 3822–3828
Russell DW, Miller AD, Alexander IE . Adeno-associated virus vectors preferentially transduce cells in S phase Proc Natl Acad Sci USA 1994 91: 8915–8919
Ponnazhagan S et al. Lack of site-specific integration of the recombinant adeno-associated virus 2 genomes in human cells Hum Gene Ther 1997 8: 275–284
Fuleihan R, Ramesh N, Geha RS . X-linked agammaglobulinemia and immunoglobulin deficiency with normal or elevated IgM: immunodeficiencies of B cell development and differentiation Adv Immunol 1995 60: 37–56
Fuleihan R et al. Cyclosporin A inhibits CD40 ligand expression in T lymphocytes J Clin Invest 1994 93: 1315–1320
Noelle RJ . CD40 and its ligand in host defense Immunity 1996 4: 415–419
Levy J et al. Clinical spectrum of X-linked hyper-IgM syndrome J Pediatr 1997 131: 47–54
Desai-Mehta A Liangjun L, Ramsey-Goldman R, Datta SK . Hyperexpression of CD40 ligand by B and T cells in humanlupus and its role in pathogenic autoantibody production J ClinInvest 1996 97: 2063–2073
Koshy M, Berger D, Crow MK . Increased expression of CD40 ligand on sytemic lupus erythematosis lymphocytes J Clin Invest 1996 98: 826–837
Clegg CH et al. Thymus dysfunction and chronic inflammatory disease in gp39 transgenic mice Int Immunol 1997 9: 1111–1122
Dunn RJ et al. Thymic overexpression of CD40 ligand disrupts normal thymic epithelial organization J Histochem Cytochem 1997 45: 129–141
Riegel JS, Corthesy B, Flanagan WM, Crabtree GR . Regulation of the interleukin-2 gene Chem Immunol 1992 51: 266–298
Jain J, Loh C, Rao A . Transcriptional regulation of the IL-2 gene Curr Opin Immunol 1995 7: 333–342
Fisher K et al. Transduction with recombinant adeno-associated virus for gene therapy is limited by leading-strand synthesis J Virol 1996 70: 520–532
Ferrari F, Samulski T, Shenk T, Samulski R . Second-strand synthesis is a rate-limiting step for efficient transduction by recombinant adeno-associated virus vectors J Virol 1996 70: 3227–3234
Nishizuka Y . The role of protein kinase C in cell surface signal transduction and tumour promotion Nature 1984 308: 693–698
Berridge MJ, Irvine RF . Inositol trisphosphate, a novel second messenger in cellular signal transduction Nature 1984 312: 315–321
Imboden JB, Stobo JD . Transmembrane signaling by the T cell antigen receptor; perturbation of the T3-antigen receptor complex generates inositol phosphates and releases calcium ions from intracellular stores J Exp Med 1985 161: 446–456
Handschumacher RE et al. Cyclophilin: a specific cytosolic binding protein for cyclosporin A Science 1984 226: 544–546
Liu J et al. Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes Cell 1991 66: 807–815
Fruman DA, Klee CB, Bierer BE, Burakoff SJ . Calcineurin phosphatase activity in T lymphocytes is inhibited by FK 506 and cyclosporin A Proc Natl Acad Sci USA 1992 89: 3686–3690
Samulski R, Chang L, Shenk T . A recombinant plasmid from which an infectious adeno-associated virus genome can be excised in vitro and its use to study viral replication J Virol 1987 61: 3096–3101
Ausubel FM et al. Preparation and analysis of DNA. In: Janssen K (ed) . Current Protocols in Molecular Biology John Wiley & Sons: New York 1995 2.0.1–2.14.8
Hsieh C-L . Basic cytogenetic techniques: culturing, slide making, and G-banding. In: Celis JE (ed) . Cell Biology: A Laboratory Handbook Academic Press: San Diego 1994 415–421
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Zhang, PX., Fuleihan, R. Transfer of activation-dependent gene expression into T cell lines by recombinant adeno-associated virus. Gene Ther 6, 182–189 (1999). https://doi.org/10.1038/sj.gt.3300803
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DOI: https://doi.org/10.1038/sj.gt.3300803
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