Abstract
Three different second-generation lentiviral self-inactivating vectors containing CMV, EF1α and PGK promoter, respectively, and all carrying the exogenous GFP gene, were compared for expression in human B-cell precursor ALL blasts. At a comparable percentage of transduction and vector DNA copy number, CMV clearly showed better efficiency of transcription. Human bone marrow stromal cells were favored compared to the MRC-5 cell line, as support for cell viability during infection. Cells were infected and analyzed after variable culture times ranging from 4 to 12 days, to reduce the possibility of pseudotransduction. In 10/14 samples, we detected more than 20% GFP-positive cells after exposure to high-titer viral supernatants. We then tested a similar vector carrying the human CD40L cDNA and, in similar infection conditions, obtained more than 20% transduction in 6/6 samples. The levels of transduction obtained were sufficient to induce the upregulation of CD83 molecule in cocultured immature dendritic cells.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Schrappe M et al. Long-term results of large prospective trials in childhood acute lymphoblastic leukemia. Leukemia 2000; 14: 2193–2194.
Schmidt-Wolf GD, Schmidt-Wolf IG . Immunomodulatory gene therapy for haematological malignancies. Br J Haematol 2002; 117: 23–32.
Stripecke R, Levine AM, Pullarkat V, Cardoso AA . Immunotherapy with acute leukemia cells modified into antigen-presenting cells: ex vivo culture and gene transfer methods. Leukemia 2002; 16: 1974–1983.
Mascarenhas L et al. Gene delivery to human B-precursor acute lymphoblastic leukemia cells. Blood 1998; 92: 3537–3545.
Stripecke R et al. Lentiviral vectors for efficient delivery of CD80 and granulocyte-macrophage-colony-stimulating factor in human acute lymphoblastic leukemia and acute myeloid leukemia cells to induce antileukemic immune responses. Blood 2000; 96: 1317–1326.
Biagi E et al. Efficient lentiviral transduction of primary human acute myelogenous and lymphoblastic leukemia cells. Haematologica 2001; 86: 13–16.
Buchschacher Jr GL, Wong-Staal F . Development of lentiviral vectors for gene therapy for human diseases. Blood 2000; 95: 2499–2504.
Zufferey R et al. Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo. Nat Biotechnol 1997; 15: 871–875.
Follenzi A et al. Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences. Nat Genet 2000; 25: 217–222.
Salmon P et al. High-level transgene expression in human hematopoietic progenitors and differentiated blood lineages after transduction with improved lentiviral vectors. Blood 2000; 96: 3392–3398.
VandenDriessche T et al. Lentiviral vectors containing the human immunodeficiency virus type-1 central polypurine tract can efficiently transduce nondividing hepatocytes and antigen-presenting cells in vivo. Blood 2002; 100: 813–822.
Bambacioni F et al. Lentiviral vectors show dramatically increased efficiency of transduction of human leukemic cell lines. Haematologica 2001; 10: 1095–1096.
Van Tendeloo VF, Van Broeckhoven C, Berneman ZN . Gene therapy: principles and applications to hematopoietic cells. Leukemia 2001; 15: 523–544.
Blomer U et al. Highly efficient and sustained gene transfer in adult neurons with a lentivirus vector. J Virol 1997; 71: 6641–6649.
Miyoshi H et al. Development of a self-inactivating lentivirus vector. J Virol 1998; 72: 8150–8157.
Case SS et al. Stable transduction of quiescent CD34(+)CD38(−) human hematopoietic cells by HIV-1-based lentiviral vectors. Proc Natl Acad Sci USA 1999; 96: 2988–2993.
Naldini L et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 1996; 272: 263–267.
Campana D et al. Human B-cell progenitors and bone marrow microenvironment. Hum Cell 1996; 9: 317–322.
Iwakuma T, Cui Y, Chang LJ . Self-inactivating lentiviral vectors with U3 and U5 modifications. Virology 1999; 261: 120–132.
Zufferey R et al. Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery. J Virol 1998; 72: 9873–9880.
Yam PY et al. Design of HIV vectors for efficient gene delivery into human hematopoietic cells. Mol Ther 2002; 5: 479–484.
Zennou V et al. HIV-1 genome nuclear import is mediated by a central DNA flap. Cell 2000; 101: 173–185.
Zufferey R, Donello JE, Trono D, Hope TJ . Woodchuck hepatitis virus posttranscriptional regulatory element enhances expression of transgenes delivered by retroviral vectors. J Virol 1999; 73: 2886–2892.
Schambach A et al. Context dependence of different modules for posttranscriptional enhancement of gene expression from retroviral vectors. Mol Ther 2000; 2: 435–445.
Ramezani A, Hawley TS, Hawley RG . Lentiviral vectors for enhanced gene expression in human hematopoietic cells. Mol Ther 2000; 2: 458–469.
Todisco E et al. CD40 ligand-stimulated B cell precursor leukemic cells elicit interferon-gamma production by autologous bone marrow T cells in childhood acute lymphoblastic leukemia. Leukemia 2002; 16: 2046–2054.
Ghia P et al. Chemoattractants MDC and TARC are secreted by malignant B-cell precursors following CD40 ligation and support the migration of leukemia-specific T cells. Blood 2001; 98: 533–540.
Cardoso AA et al. Ex vivo generation of human anti-pre-B leukemia-specific autologous cytolytic T cells. Blood 1997; 90: 549–561.
Cardoso AA et al. Pre-B acute lymphoblastic leukemia cells may induce T-cell anergy to alloantigen. Blood 1996; 88: 41–48.
Ranheim EA, Kipps TJ . Activated T cells induce expression of B7/BB1 on normal or leukemic B cells through a CD40-dependent signal. J Exp Med 1993; 177: 925–935.
Schultze JL et al. CD40-activated human B cells: an alternative source of highly efficient antigen presenting cells to generate autologous antigen-specific T cells for adoptive immunotherapy. J Clin Invest 1997; 100: 2757–2765.
Van den Hove LE et al. CD40 triggering of chronic lymphocytic leukemia B cells results in efficient alloantigen presentation and cytotoxic T lymphocyte induction by up-regulation of CD80 and CD86 costimulatory molecules. Leukemia 1997; 11: 572–580.
Kato K, Cantwell MJ, Sharma S, Kipps TJ . Gene transfer of CD40-ligand induces autologous immune recognition of chronic lymphocytic leukemia B cells. J Clin Invest 1998; 101: 1133–1141.
Takahashi S et al. Autologous antileukemic immune response induced by chronic lymphocytic leukemia B cells expressing the CD40 ligand and interleukin 2 transgenes. Hum Gene Ther 2001; 12: 659–670.
Dotti G et al. Transgenic expression of CD40 ligand produces an in vivo antitumor immune response against both CD40(+) and CD40(−) plasmacytoma cells. Blood 2002; 100: 200–207.
Dotti G et al. Adenovector-induced expression of human-CD40-ligand (hCD40L) by multiple myeloma cells. A model for immunotherapy. Exp Hematol 2001; 29: 952–961.
Gruber TA, Skelton DC, Kohn DB . Requirement for NK cells in CD40 ligand-mediated rejection of Philadelphia chromosome-positive acute lymphoblastic leukemia cells. J Immunol 2002; 168: 73–80.
Dilloo D et al. CD40 ligand induces an antileukemia immune response in vivo. Blood 1997; 90: 1927–1933.
Saudemont A et al. Gene transfer of CD154 and IL12 cDNA induces an anti-leukemic immunity in a murine model of acute leukemia. Leukemia 2002; 16: 1637–1644.
Vereecque R et al. Gene transfer of GM-CSF, CD80 and CD154 cDNA enhances survival in a murine model of acute leukemia with persistence of a minimal residual disease. Gene Therapy 2000; 7: 1312–1316.
Urashima M et al. An oral CD40 ligand gene therapy against lymphoma using attenuated Salmonella typhimurium. Blood 2000; 95: 1258–1263.
Wierda WG et al. CD40-ligand (CD154) gene therapy for chronic lymphocytic leukemia. Blood 2000; 96: 2917–2924.
Tolba KA et al. Development of herpes simplex virus-1 amplicon-based immunotherapy for chronic lymphocytic leukemia. Blood 2001; 98: 287–295.
Kuwashima N, Kageyama S, Eto Y, Urashima M . CD40 ligand immunotherapy in cancer: an efficient approach. Leuk Lymphoma 2001; 42: 1367–1377.
Loskog A, Totterman TH, Bohle A, Brandau S . In vitro activation of cancer patient-derived dendritic cells by tumor cells genetically modified to express CD154. Cancer Gene Ther 2002; 9: 846–853.
Wendtner CM et al. Efficient gene transfer of CD40 ligand into primary B-CLL cells using recombinant adeno-associated virus (rAAV) vectors. Blood 2002; 100: 1655–1661.
Makrynikola V, Bradstock KF . Adhesion of precursor-B acute lymphoblastic leukaemia cells to bone marrow stromal proteins. Leukemia 1993; 7: 86–92.
Sastry L et al. Titering lentiviral vectors: comparison of DNA, RNA and marker expression methods. Gene Therapy 2002; 9: 1155–1162.
Pongers-Willemse et al. Realtime quantitative PCR for the detection of minimal residual disease in acute lymphoblastic leukemia using junctional region specific TaqMan probes. Leukemia 1998; 12: 2006–2014.
Albert DM et al. Establishment of cell lines of uveal melanoma. Methodology and characteristics. Invest Ophthalmol Vis Sci 1984; 25: 1284–1299.
Franco S et al. Clonal variation in phenotype and life span of human embryonic fibroblasts (MRC-5) transduced with the catalytic component of telomerase (hTERT). Exp Cell Res 2001; 268: 14–25.
Gallardo HF, Tan C, Ory D, Sadelain M . Recombinant retroviruses pseudotyped with the vesicular stomatitis virus G glycoprotein mediate both stable gene transfer and pseudotransduction in human peripheral blood lymphocytes. Blood 1997; 90: 952–957.
Liu ML, Winther BL, Kay MA . Pseudotransduction of hepatocytes by using concentrated pseudotyped vesicular stomatitis virus G glycoprotein (VSV-G)-Moloney murine leukemia virus-derived retrovirus vectors: comparison of VSV-G and amphotropic vectors for hepatic gene transfer. J Virol 1996; 70: 2497–2502.
Haas DL, Case SS, Crooks GM, Kohn DB . Critical actors influencing stable transduction of human CD34(+) cells with HIV-1-derived lentiviral vectors. Mol Ther 2000; 2: 71–80.
Min WS, Song CW, Uckun FM . Thermal sensitivity and thermal tolerance of human B-lineage acute lymphoblastic leukemia (ALL) cells. Int J Radiat Oncol Biol Phys 1990; 18: 147–153.
Yeoh EJ et al. Classification, subtype discovery, and prediction of outcome in pediatric acute lymphoblastic leukemia by gene expression profiling. Cancer Cell 2002; 1: 133–143.
Takahashi S et al. Transgenic expression of CD40L and interleukin-2 induces an autologous antitumor immune response in patients with non-Hodgkin's lymphoma. Cancer Gene Ther 2001; 8: 378–387.
Kipps TJ, Chu P, Wierda WG . Immunogenetic therapy for B-cell malignancies. Semin Oncol 2000; 27: 104–109.
Bleharski JR et al. Signaling lymphocytic activation molecule is expressed on CD40 ligand-activated dendritic cells and directly augments production of inflammatory cytokines. J Immunol 2001; 167: 3174–3181.
Dull T et al. A third-generation lentivirus vector with a conditional packaging system. J Virol 1998; 72: 8463–8471.
Koya RC et al. Transduction of acute myeloid leukemia cells with third generation self-inactivating lentiviral vectors expressing CD80 and GM-CSF: effects on proliferation, differentiation, and stimulation of allogeneic and autologous anti-leukemia immune responses. Leukemia 2002; 16: 1645–1654.
Zaiss AK, Son S, Chang LJ . RNA 3' readthrough of oncoretrovirus and lentivirus: implications for vector safety and efficacy. J Virol 2002; 76: 7209–7219.
Powell SK et al. In vitro analysis of transformation potential associated with retroviral vector insertions. Hum Gene Ther 1999; 10: 2123–2132.
Bukovsky AA, Song JP, Naldini L . Interaction of human immunodeficiency virus-derived vectors with wild-type virus in transduced cells. J Virol 1999; 73: 7087–7092.
Manganini M et al. A human immunodeficiency virus type 1 pol gene-derived sequence (cPPT/CTS) increases the efficiency of transduction of human nondividing monocytes and T lymphocytes by lentiviral vectors. Hum Gene Ther 2002; 13: 1793–1807.
Limon A et al. High-titer retroviral vectors containing the enhanced green fluorescent protein gene for efficient expression in hematopoietic cells. Blood 1997; 90: 3316–3321.
Manabe A et al. Bone marrow-derived stromal cells prevent apoptotic cell death in B-lineage acute lymphoblastic leukemia. Blood 1992; 79: 2370–2377.
Campana D, Manabe A, Evans WE . Stroma-supported immunocytometric assay (SIA): a novel method for testing the sensitivity of acute lymphoblastic leukemia cells to cytotoxic drugs. Leukemia 1993; 7: 482–488.
Kumagai M et al. Stroma-supported culture in childhood B-lineage acute lymphoblastic leukemia cells predicts treatment outcome. J Clin Invest 1996; 97: 755–760.
Allavena P et al. IL-10 prevents the differentiation of monocyte to dendritic cells but promotes their maturation to macrophages. Eur J Immunol 1998; 28: 359–369.
Sallusto F, Lanzavecchia A . Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor-alpha. J Exp Med 1994; 179: 1109–1118.
Acknowledgements
This work was supported in part by grants from the Associazione Italiana Ricerca sul Cancro (to MI and AB), the Istituto Superiore di Sanità (ISS, Rome, AIDS grants 30C.40, 30D.41 and 40C.66 to MI), the Italian Ministry for University and Research (project FIRB no. RBAU01J2ER and RBAU01H8SX to MI), the Fondazione Tettamanti, Monza, the MIUR – Oncologia (to AB) and Progetti di Ricerca Finalizzata Ministero della Salute RF202/02 (to AB and MB). MB was supported by fellowships from the Fundaçao Ary Frauzino (FAF, Brazil), the Fundaçao PROVITA (Brazil) and the Coordenaçao de Aperfeiçoamento de Pessoal de Nivel Superior (CAPES, Brazil). GD'A was supported by a grant from an FIRC (Fondazione Italiana Ricerca Cancro) fellowship.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bonamino, M., Serafini, M., D'Amico, G. et al. Functional transfer of CD40L gene in human B-cell precursor ALL blasts by second-generation SIN lentivectors. Gene Ther 11, 85–93 (2004). https://doi.org/10.1038/sj.gt.3302141
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3302141
Keywords
This article is cited by
-
Human mesenchymal stromal/stem cells recruit resident pericytes and induce blood vessels maturation to repair experimental spinal cord injury in rats
Scientific Reports (2020)
-
Transposon-mediated generation of CAR-T cells shows efficient anti B-cell leukemia response after ex vivo expansion
Gene Therapy (2020)
-
PCA3 long noncoding RNA modulates the expression of key cancer-related genes in LNCaP prostate cancer cells
Tumor Biology (2016)
-
T cells stimulated by CD40L positive leukemic blasts-pulsed dendritic cells meet optimal functional requirements for adoptive T-cell therapy
Leukemia (2006)
-
Epstein–Barr virus vector-mediated gene transfer into human B cells: potential for antitumor vaccination
Gene Therapy (2006)