Abstract
Gene silencing with RNAi is an invaluable technique in cell biology to knock down the target gene expression. Dendritic cells (DC) are the most effective antigen-presenting cells (APC), and the efficacy of antigen presentation is tightly controlled by the stimulatory as well as inhibitory mechanisms. In recent studies, RNAi technology has been employed to silence the expression of the intrinsic inhibitors of antigen presentation in DC, improving the efficacy of DC vaccines against tumor antigens in pre-clinical studies. Here, we describe the technique of using siRNA oligonucleotides, adenovirus expressing shRNA (Ad-shRNA), or lentivirus expressing shRNA (Lv-shRNA) to knock down inhibitors of antigen presentation in both mouse and human DC.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Rosenberg SA et al (2004) Cancer immunotherapy: moving beyond current vaccines. Nat Med 10:909–915
Iwasaki A, Medzhitov R (2004) Toll-like receptor control of the adaptive immune responses. Nat Immunol 5(10):987–995
Kaisho T, Akira S (2003) Regulation of dendritic cell function through Toll-like receptors. Curr Mol Med 3(4):373–385
Kubo T et al (2004) Regulatory T cell suppression and anergy are differentially regulated by proinflammatory cytokines produced by TLR-activated dendritic cells. J Immunol 173:7249–7258
Goldstein DR (2004) Toll-like receptors and other links between innate and acquired alloimmunity. Curr Opin Immunol 16(5):538–544
Pasare C, Medzhitov R (2003) Toll pathway-dependent blockade of CD4 + CD25+ T cell-mediated suppression by dendritic cells. Science 299:1033–1036
Lee EG et al (2000) Failure to regulate TNF-induced NF-kappaB and cell death responses in A20-deficient mice. Science 289:2350–2354
Boone DL et al (2004) The ubiquitin-modifying enzyme A20 is required for termination of Toll-like receptor responses. Nat Immunol 5:1052–1060
Wertz IE et al (2004) De-ubiquitination and ubiquitin ligase domains of A20 downregulate NF-kappaB signalling. Nature 430:694–699
Sarma V et al (1995) Activation of the B-cell surface receptor CD40 induces A20, a novel zinc finger protein that inhibits apoptosis. J Biol Chem 270:12343–12346
Saitoh T et al (2005) A20 is a negative regulator of IFN regulatory factor 3 signaling. J Immunol 174:1507–1512
Song XT et al (2008) A20 is an antigen presentation attenuator, and its inhibition overcomes regulatory T cell-mediated suppression. Nat Med 14:258–265
Hong B et al (2011) Mucosal and systemic anti-HIV immunity controlled by A20 in mouse dendritic cells. J Clin Invest 121(2):739–751
Song XT et al (2011) A Th1-inducing adenoviral vaccine for boosting adoptively transferred T cells. Mol Ther 19(1):211–217
Hong B et al (2009) Human suppressor of cytokine signaling 1 controls immunostimulatory activity of monocyte-derived dendritic cells. Cancer Res 69(20):8076–8084
Breckpo K et al (2009) Attenuated expression of A20 markedly increases the efficacy of double-stranded RNA-activated dendritic cells as an anti-cancer vaccine. J Immunol 182(2):860–870
Acknowledgement
This work was supported by NIH grant R01 CA148748 and DOD W81XWH-10-1-0281.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Song, XT. (2014). Genetic Modification of Dendritic Cells with RNAi. In: Lawman, M., Lawman, P. (eds) Cancer Vaccines. Methods in Molecular Biology, vol 1139. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0345-0_11
Download citation
DOI: https://doi.org/10.1007/978-1-4939-0345-0_11
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0344-3
Online ISBN: 978-1-4939-0345-0
eBook Packages: Springer Protocols