Skip to main content

Immunity of Lentiviral Vector-Modified Dendritic Cells

  • Protocol
  • First Online:
Gene Therapy of Cancer

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 542))

Summary

Innovative approaches to induce a strong immune response are key to the success of immunotherapy. Dendritic cells (DCs) are professional antigen-presenting cells (APCs) equipped with co-stimulatory, adhesion, and major histocompatibility complex (MHC) molecules needed for initiation and reactivation of the immune response. DCs are able to initiate and stimulate both innate and adaptive immune responses and, by secretion of cytokines, chemokines, and expression of regulatory molecules, to shape the adaptive immune response toward a long-lasting memory immunity. DCs from the peripheral blood of immune-compromised patients, however, often display an immature phenotype with defective functions. This emphasizes the importance and potential of engineering antigen-specific DCs in vitro. A state-of-the-art approach to overcome the prevailing immune dysfunction(s) in patients is to engineer DCs or DC progenitors to generate fully functional DCs for the modification of host immunity. Lentiviral vectors (LVs) are highly efficient gene transfer vehicles for engineering DC functions. Examples of lentiviral vectors encoding immune-modulatory genes and useful functional assays for the analysis of effector immune cell response are described in this chapter.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Guermonprez P, Valladeau J, Zitvogel L, Thery C, Amigorena S. (2002) Antigen presentation and T cell stimulation by dendritic cells. Annu Rev Immunol 20:621–667.

    Article  PubMed  CAS  Google Scholar 

  2. Condon C, Watkins SC, Celluzzi CM, Thompson K, Falo LDJ. (1996) DNA-based immunization by in vivo transfection of dendritic cells. Nat Med 2:1122–1128.

    Article  PubMed  CAS  Google Scholar 

  3. Liu M. (1998) Transfected human dendritic cells as cancer vaccines. Nat Biotechnol 16:335–336.

    Article  PubMed  CAS  Google Scholar 

  4. Chen X, He J, Chang L-J. (2004) Alteration of T cell immunity by lentiviral transduction of human monocyte-derived dendritic cells. Retrovirology 1:37.

    Article  PubMed  Google Scholar 

  5. Wang B, He J, Liu C, Chang LJ. (2006) An effective cancer vaccine modality: Lentiviral modification of dendritic cells expressing multiple cancer-specific antigens. Vaccine 24:3477–3489.

    Article  PubMed  CAS  Google Scholar 

  6. Ludewig B, Ehl S, Karrer U, Odermatt B, Hengartner H, Zinkernagel RM. (1998) Dendritic cells efficiently induce protective antiviral immunity. J Virol 72:3812–3818.

    PubMed  CAS  Google Scholar 

  7. Kirk CJ, Mule JJ. (2000) Gene-modified dendritic cells for use in tumor vaccines. Hum Gene Ther 11:797–806.

    Article  PubMed  CAS  Google Scholar 

  8. Jenne L, Schuler G, Steinkasserer A. (2001) Viral vectors for dendritic cell-based immunotherapy. Trends Immunol 22:102–107.

    Article  PubMed  CAS  Google Scholar 

  9. Murphy A, Westwood JA, Teng MW, Moeller M, Darcy PK, Kershaw MH. (2005) Gene modification strategies to induce tumor immunity. Immunity 22:403–414.

    Article  PubMed  CAS  Google Scholar 

  10. Breckpot K, Aerts JL, Thielemans K. (2007) Lentiviral vectors for cancer immunotherapy: transforming infectious particles into therapeutics. Gene Ther 14:847–862.

    Article  PubMed  CAS  Google Scholar 

  11. Lipscomb MF, Masten BJ. (2002) Dendritic cells: immune regulators in health and disease. Physiol Rev 82:97–130.

    PubMed  CAS  Google Scholar 

  12. Mosmann TR, Coffman RL. (1989) TH1 and TH2 cells: different patterns of lymphokine secretion lead to different functional properties. Annu Rev Immunol 7:145–173.

    Article  PubMed  CAS  Google Scholar 

  13. Kalinski P, Hilkens CM, Wierenga EA, Kapsenberg ML. (1999) T-cell priming by type-1 and type-2 polarized dendritic cells: the concept of a third signal. Immunol Today 20:561–567.

    Article  PubMed  CAS  Google Scholar 

  14. Chambers CA. (2001) The expanding world of co-stimulation: the two-signal model revisited. Trends Immunol 22:217–223.

    Article  PubMed  CAS  Google Scholar 

  15. Chirathaworn C, Kohlmeier JE, Tibbetts SA, Rumsey LM, Chan MA, Benedict SH. (2002) Stimulation through intercellular adhesion molecule-1 provides a second signal for T cell activation. J Immunol 168:5530–5537.

    PubMed  CAS  Google Scholar 

  16. Salomon B, Bluestone JA. (1998) LFA-1 interaction with ICAM-1 and ICAM-2 regulates Th2 cytokine production. J Immunol 161:5138–5142.

    PubMed  CAS  Google Scholar 

  17. Sabatte J, Maggini J, Nahmod K, Amaral MM, Martinez D, Salamone G, Ceballos A,. (2007) Interplay of pathogens, cytokines and other stress signals in the regulation of dendritic cell function. Cytokine Growth Factor Rev 18:5–17.

    Article  PubMed  CAS  Google Scholar 

  18. Han S, Wang B, Cotter MJ, Yang LJ, Zucali J, Moreb JS, Chang LJ. (2008) Overcoming Immune Tolerance Against Multiple Myeloma With Lentiviral Calnexin-engineered Dendritic Cells. Mol Ther 16: 269–279.

    Article  PubMed  CAS  Google Scholar 

  19. Chang L-J, Zaiss A-K. (2001) Methods for the preparation and use of lentivirus vectors. In: Morgan J, ed. Gene Therapy Protocols, Vol. 2, 2nd ed. Humana Press, Totowa, NJ, 303–318.

    Chapter  Google Scholar 

  20. Chang L-J, Zaiss A-K. (2001) Self inactivating lentiviral vectors in combination with a sensitive Cre/loxP reporter system. In: Walker J, ed. Methods in Molecular Medicine. Humana Press, Totowa, NJ, 367–382.

    Google Scholar 

  21. Lyons AB, Parish CR. (1994) Determination of lymphocyte division by flow cytometry. J Immunol Methods 171:131–137.

    Article  PubMed  CAS  Google Scholar 

  22. Jedema I, van der Werff NM, Barge RM, Willemze R, Falkenburg JH. (2004) New CFSE-based assay to determine susceptibility to lysis by cytotoxic T cells of leukemic precursor cells within a heterogeneous target cell population. Blood 103:2677–2682.

    Article  PubMed  CAS  Google Scholar 

  23. Betts MR, Brenchley JM, Price DA, De Rosa SC, Douek DC, Roederer M, Koup RA. (2003) Sensitive and viable identification of antigen-specific CD8 + T cells by a flow cytometric assay for degranulation. J Immunol Methods 281:65–78.

    Article  PubMed  CAS  Google Scholar 

  24. Hopken UE, Lehmann I, Droese J, Lipp M, Schuler T, Rehm A. (2005) The ratio between dendritic cells and T cells determines the outcome of their encounter: proliferation versus deletion. Eur J Immunol 35:2851–2863.

    Article  PubMed  Google Scholar 

  25. Mannering SI, Morris JS, Jensen KP, Purcell AW, Honeyman MC, van Endert PM, Harrison LC. (2003) A sensitive method for detecting proliferation of rare autoantigen-specific human T cells. J Immunol Methods 283:173–183.

    Article  PubMed  CAS  Google Scholar 

  26. Denkberg G, Cohen CJ, Reiter Y. (2001) Critical role for CD8 in binding of MHC tetramers to TCR: CD8 antibodies block specific binding of human tumor-specific MHC-peptide tetramers to TCR. J Immunol 167:270–276.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Lung-Ji Chang .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2009 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Han, S., Chang, LJ. (2009). Immunity of Lentiviral Vector-Modified Dendritic Cells. In: Walther, W., Stein, U. (eds) Gene Therapy of Cancer. Methods in Molecular Biology™, vol 542. Humana Press. https://doi.org/10.1007/978-1-59745-561-9_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-561-9_13

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-934115-85-5

  • Online ISBN: 978-1-59745-561-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics