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High-titer lentiviral vectors stimulate fetal calf serum-specific human CD4 T-cell responses: implications in human gene therapy

Gene Therapy volume 16, pages 788–795 (2009)Cite this article

Abstract

Human immunodeficiency virus-1-derived lentiviral vectors have been increasingly used for gene delivery in both pre-clinical and clinical models. Numerous studies have shown that dendritic cells (DC) transduced with concentrated lentiviral vectors can induce primary T-cell responses to viral and tumor antigens. In this study, we attempted to generate influenza hemagglutinin-specific CD4 T cells using lentiviral vectors containing the signal sequence and human lysosome-associated membrane protein to target hemagglutinin to the major histocompatibility complex class II processing pathway. Autologous dendritic cells were generated in serum-free medium and transduced with concentrated, high-titer lentiviruses to stimulate autologous T cells. Unexpectedly, we failed to generate influenza hemagglutinin-specific CD4 T cells rather than T cells specific for fetal calf serum (FCS). By limiting dilution, we established several FCS-specific CD4 T-cell clones restricted by human leukocyte antigen-DR1 and human leukocyte antigen-DR4. Lentiviruses produced in human serum-adapted 293 cells or in serum-free medium were unable to sensitize dendritic cells for recognition by FCS-specific CD4 T-cell clones. Our results indicate that residual FCS in concentrated lentiviral pellets is, in part, responsible for its immunogenicity. These FCS-specific CD4 T cells may be useful in testing clinical grade lentiviral vectors for the presence of contaminating FCS.

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References

  1. Kordower JH, Emborg ME, Bloch J, Ma SY, Chu Y, Leventhal L et al. Neurodegeneration. Science 2000; 290: 767–773.

    Article  CAS  PubMed  Google Scholar 

  2. Consiglio A, Quattrini A, Martino S, Bensadoun JC, Dolcetta D, Trojani A et al. In vivo gene therapy of metachromatic leukodystrophy by lentiviral vectors: correction of neuropathology and protection against learning impairments in affected mice. Nat Med 2001; 7: 310–316.

    Article  CAS  PubMed  Google Scholar 

  3. Azzouz M, Le T, Ralph GS, Walmsley L, Monani UR, Lee DC et al. Lentivector-mediated SMN replacement in a mouse model of spinal muscular atrophy. J Clin Invest 2004; 114: 1726–1731.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Levine BL, Humeau LM, Boyer J, MacGregor RR, Rebello T, Lu X et al. Gene transfer in humans using a conditionally replicating lentiviral vector. Proc Natl Acad Sci USA 2006; 103: 17372–17377.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Naldini L, Blomer U, Gage FH, Trono D, Verma IM . Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector. Proc Natl Acad Sci USA 1996; 93: 11382–11388.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Naldini L, Blomer U, Gallay P, Ory D, Mulligan R, Gage FH et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 1996; 272: 263–267.

    Article  CAS  PubMed  Google Scholar 

  7. Saenz DT, Poeschla EM . FIV: from lentivirus to lentivector. J Gene Med 2004; 6: S95–104.

    Article  CAS  PubMed  Google Scholar 

  8. Blomer U, Naldini L, Kafri T, Trono D, Verma IM, Gage FH . Highly efficient and sustained gene transfer in adult neurons with a lentivirus vector. J Virol 1997; 71: 6641–6649.

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D et al. A third- generation lentivirus vector with a conditional packaging system. J Virol 1998; 72: 8463–8471.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Zufferey R, Nagy D, Mandel RJ, Naldini L, Trono D . Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo. Nat Biotech 1997; 15: 871–875.

    Article  CAS  Google Scholar 

  11. Zufferey R, Dull T, Mandel RJ, Bukovsky A, Quiroz D, Naldini L et al. Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery. J Virol 1998; 72: 9873–9880.

    CAS  PubMed  PubMed Central  Google Scholar 

  12. van Lunzen J, Glaunsinger T, Stahmer I, von Baehr V, Baum C, Schilz A et al. Transfer of autologous gene-modified T cells in HIV-infected patients with advanced immunodeficiency and drug-resistant virus. Mol Ther 2007; 15: 1024–1033.

    Article  CAS  PubMed  Google Scholar 

  13. Gruber A, Kan-Mitchell J, Kuhen KL, Mukai T, Wong-Staal F . Dendritic cells transduced by multiply deleted HIV-1 vectors exhibit normal phenotypes and functions and elicit an HIV-specific cytotoxic T-lymphocyte response in vitro. Blood 2000; 96: 1327–1333.

    CAS  PubMed  Google Scholar 

  14. Dyall J, Latouche J-B, Schnell S, Sadelain M . Lentivirus-transduced human monocyte-derived dendritic cells efficiently stimulate antigen-specific cytotoxic T lymphocytes. Blood 2001; 97: 114–121.

    Article  CAS  PubMed  Google Scholar 

  15. Firat H, Zennou V, Garcia-Pons F, Ginhoux F, Cochet M, Danos O et al. Use of a lentiviral flap vector for induction of CTL immunity against melanoma. Perspectives for immunotherapy. J Gene Med 2002; 4: 38–45.

    Article  PubMed  Google Scholar 

  16. Breckpot K, Dullaers M, Bonehill A, Van Meirvenne S, Heirman C, De Greef C et al. Lentivirally transduced dendritic cells as a tool for cancer immunotherapy. J Gene Med 2003; 5: 654–667.

    Article  CAS  PubMed  Google Scholar 

  17. Lizee G, Gonzales MI, Topalian SL . Lentivirus vector-mediated expression of tumor-associated epitopes by human antigen presenting cells. Hum Gene Ther 2004; 15: 393–404.

    Article  CAS  PubMed  Google Scholar 

  18. Chen X, Wang B, Chang L-J . Induction of primary anti-HIV CD4 and CD8 T cell responses by dendritic cells transduced with self-inactivating lentiviral vectors. Cell Immunol 2006; 243: 10–18.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Breckpot K, Aerts JL, Thielemans K . Lentiviral vectors for cancer immunotherapy: transforming infectious particles into therapeutics. Gene Therapy 2007; 14: 847–862.

    Article  CAS  PubMed  Google Scholar 

  20. Wu TC, Guarnieri FG, Staveley-O’Carroll KF, Viscidi RP, Levitsky HI, Hedrick L et al. Engineering an intracellular pathway for major histocompatibility complex class II presentation of antigens. Proc Natl Acad Sci USA 1995; 92: 11671–11675.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Riddell SR, Elliott M, Lewinsohn DA, Gilbert MJ, Wilson L, Manley SA et al. T-cell mediated rejection of gene-modified HIV-specific cytotoxic T lymphocytes in HIV-infected patients. Nat Med 1996; 2: 216–223.

    Article  CAS  PubMed  Google Scholar 

  22. Bonini C, Ferrari G, Verzeletti S, Servida P, Zappone E, Ruggieri L et al. HSV-TK gene transfer into donor lymphocytes for control of allogeneic graft-versus-leukemia. Science 1997; 276: 1719–1724.

    Article  CAS  PubMed  Google Scholar 

  23. Selvaggi TA, Walker RE, Fleisher TA . Development of antibodies to fetal calf serum with arthus-like reactions in human immunodeficiency virus-infected patients given syngeneic lymphocyte infusions. Blood 1997; 89: 776–779.

    CAS  PubMed  Google Scholar 

  24. Tuschong L, Soenen SL, Blaese RM, Candotti F, Muul LM . Immune response to fetal calf serum by two adenosine deaminase-deficient patients after T cell gene therapy. Hum Gene Ther 2002; 13: 1605–1610.

    Article  CAS  PubMed  Google Scholar 

  25. Mackensen A, Drager R, Schlesier M, Mertelsmann R, Lindemann A . Presence of IgE antibodies to bovine serum albumin in a patient developing anaphylaxis after vaccination with human peptide-pulsed dendritic cells. Cancer Immunol Immunother 2000; 49: 152–156.

    Article  CAS  PubMed  Google Scholar 

  26. Baekelandt V, Claeys A, Eggermont K, Lauwers E, De Strooper B, Nuttin B et al. Characterization of lentiviral vector-mediated gene transfer in adult mouse brain. Hum Gene Ther 2002; 13: 841–853.

    Article  CAS  PubMed  Google Scholar 

  27. Baekelandt V, Eggermont K, Michiels M, Nuttin B, Debyser Z . Optimized lentiviral vector production and purification procedure prevents immune response after transduction of mouse brain. Gene Therapy 2003; 10: 1933–1940.

    Article  CAS  PubMed  Google Scholar 

  28. Wang RF, Wang X, Atwood AC, Topalian SL, Rosenberg SA . Cloning genes encoding MHC class II-restricted antigens: mutated CDC27 as a tumor antigen. Science 1999; 284: 1351–1354.

    Article  CAS  PubMed  Google Scholar 

  29. Amendola M, Venneri MA, Biffi A, Vigna E, Naldini L . Coordinate dual-gene transgenesis by lentiviral vectors carrying synthetic bidirectional promoters. Nat Biotechnol 2005; 23: 108–116.

    Article  CAS  PubMed  Google Scholar 

  30. Zhou X, Cui Y, Huang X, Yu Z, Thomas AM, Ye Z et al. Lentivirus-mediated gene transfer and expression in established human tumor antigen-specific cytotoxic T cells and primary unstimulated T cells. Hum Gene Ther 2003; 14: 1089–1105.

    Article  CAS  PubMed  Google Scholar 

  31. Tiscornia G, Singer O, Verma IM . Production and purification of lentiviral vectors. Nat Protocols 2006; 1: 241–245.

    Article  CAS  PubMed  Google Scholar 

  32. Riddell SR, Greenberg PD . The use of anti-CD3 and anti-CD28 monoclonal antibodies to clone and expand human antigen-specific T cells. J Immunol Methods 1990; 128: 189–201.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We are grateful to Dr Nikunj Somia for helpful discussion on lentiviral purification and Dr Nicola Philpott for English editing. LB was a recipient of a Baxter Oncology Postdoctoral Fellowship of the Amy Strelzer Manasevit Scholars Program, the National Marrow Donor Program. XZ was a recipient of an American Society of Hematology Junior Faculty Award. This work was supported by the Children's Cancer Research Fund in Minneapolis, Viking Children's Fund and the University of Minnesota Masonic Cancer Center (XZ). LB and HG designed, carried out the research, analyzed the data and wrote the paper; XH, ST and MW designed, carried out the research, analyzed the data and edited the paper; RSM discussed the work and wrote the paper; XZ oversaw the work and wrote the paper.

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Author notes

  1. L Bao and H Guo: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pediatrics, Division of Blood and Marrow Transplantation, University of Minnesota Medical School, Minneapolis, MN, USA

    L Bao, H Guo, X Huang, M Wong & X Zhou

  2. Masonic Cancer Center, University of Minnesota Medical School, Minneapolis, MN, USA

    L Bao, H Guo, X Huang, M Wong, R S McIvor & X Zhou

  3. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA

    L Bao, H Guo, X Huang, M Wong & X Zhou

  4. Center for Genome Engineering,

    L Bao, H Guo, X Huang, M Wong, R S McIvor & X Zhou

  5. , University of Minnesota Medical School, Minneapolis, MN, USA

    L Bao, H Guo, X Huang, M Wong, R S McIvor & X Zhou

  6. Graduate Program in Microbial Engineering, University of Minnesota Graduate School, Minneapolis, MN, USA

    S Tammana & X Zhou

  7. Department of Genetics, Cell Biology and Development, University of Minnesota Medical School, Minneapolis, MN, USA

    R S McIvor

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  1. L Bao
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Correspondence to X Zhou.

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Bao, L., Guo, H., Huang, X. et al. High-titer lentiviral vectors stimulate fetal calf serum-specific human CD4 T-cell responses: implications in human gene therapy. Gene Ther 16, 788–795 (2009). https://doi.org/10.1038/gt.2009.34

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  • DOI: https://doi.org/10.1038/gt.2009.34

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