Abstract
The importance of tumor-associated antigen-specific T cells in the effective control of cancer has been highlighted by recent advances in cancer immunotherapies that target the programmed cell death-1 (PD-1) pathway or that utilize modified T cell receptors. Phosphopeptide-specific T cells are of interest because they recognize a new class of tumor antigens that are derived from proteins relevant for cancer development and growth. These T cell lines or their antigen receptors can be used in combination with other forms of therapy to improve the immune response and survival of cancer patients. We describe here a protocol for the generation of human and transgenic murine phosphopeptide-specific T cells lines as tools for investigating T cell reactivity against melanoma phosphoantigens displayed by HLA-A*0201.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Zarling AL, Ficarro SB, White FM, Shabanowitz J, Hunt DF, Engelhard VH (2000) Phosphorylated peptides are naturally processed and presented by major histocompatibility complex class I molecules in vivo. J Exp Med 192(12):1755–1762. https://doi.org/10.1084/jem.192.12.1755
Zarling AL, Polefrone JM, Evans AM, Mikesh LM, Shabanowitz J, Lewis ST, Engelhard VH, Hunt DF (2006) Identification of class I MHC-associated phosphopeptides as targets for cancer immunotherapy. Proc Natl Acad Sci U S A 103(40):14889–14894. https://doi.org/10.1073/pnas.0604045103
Depontieu FR, Qian J, Zarling AL, McMiller TL, Salay TM, Norris A, English AM, Shabanowitz J, Engelhard VH, Hunt DF, Topalian SL (2009) Identification of tumor-associated, MHC class II-restricted phosphopeptides as targets for immunotherapy. Proc Natl Acad Sci U S A 106(29):12073–12078. https://doi.org/10.1073/pnas.0903852106
Ferguson AR, Nichols LA, Zarling AL, Thompson ED, Brinkman CC, Hargadon KM, Bullock TN, Engelhard VH (2008) Strategies and challenges in eliciting immunity to melanoma. Immunol Rev 222:28–42. https://doi.org/10.1111/j.1600-065X.2008.00620.x
Cobbold M, De La Peña H, Norris A, Polefrone JM, Qian J, English AM, Cummings KL, Penny S, Turner JE, Cottine J, Abelin JG, Malaker SA, Zarling AL, Huang HW, Goodyear O, Freeman SD, Shabanowitz J, Pratt G, Craddock C, Williams ME, Hunt DF, Engelhard VH (2013) MHC class I-associated phosphopeptides are the targets of memory-like immunity in leukemia. Sci Transl Med 5(203):203ra125. https://doi.org/10.1126/scitranslmed.3006061
Zarling AL, Obeng RC, Desch AN, Pinczewski J, Cummings KL, Deacon DH, Conaway M, Slingluff CL, Engelhard VH (2014) MHC-restricted phosphopeptides from insulin receptor substrate-2 and CDC25b offer broad-based immunotherapeutic agents for cancer. Cancer Res 74(23):6784–6795. https://doi.org/10.1158/0008-5472.CAN-14-0043
Li Y, Depontieu FR, Sidney J, Salay TM, Engelhard VH, Hunt DF, Sette A, Topalian SL, Mariuzza RA (2010) Structural basis for the presentation of tumor-associated MHC class II-restricted phosphopeptides to CD4+ T cells. J Mol Biol 399(4):596–603. https://doi.org/10.1016/j.jmb.2010.04.037
Mohammed F, Cobbold M, Zarling AL, Salim M, Barrett-Wilt GA, Shabanowitz J, Hunt DF, Engelhard VH, Willcox BE (2008) Phosphorylation-dependent interaction between antigenic peptides and MHC class I: a molecular basis for the presentation of transformed self. Nat Immunol 9(11):1236–1243. https://doi.org/10.1038/ni.1660
Mohammed F, Stones DH, Zarling AL, Willcox CR, Shabanowitz J, Cummings KL, Hunt DF, Cobbold M, Engelhard VH, Willcox BE (2017) The antigenic identity of human class I MHC phosphopeptides is critically dependent upon phosphorylation status. Oncotarget 8(33):54160–54172. https://doi.org/10.18632/oncotarget.16952
Engelhard VH, Obeng RC, Cummings KL, Petroni GR, Ambakhutwala AL, Chianese-Bullock KA, Smith KT, Lulu A, Varhegyi N, Smolkin ME, Myers P, Mahoney KE, Shabanowitz J, Buettner N, Hall EH, Haden K, Cobbold M, Hunt DF, Weiss G, Gaughan E, Slingluff CL Jr (2020) MHC-restricted phosphopeptide antigens: preclinical validation and first-in-humans clinical trial in participants with high-risk melanoma. J Immunother Cancer 8(1):e000262. https://doi.org/10.1136/jitc-2019-000262
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Obeng, R.C., Ambakhutwala, A.L. (2021). Generation of Phosphopeptide-Specific T Cell Lines as Tools for Melanoma Immunotherapy. In: Hargadon, K.M. (eds) Melanoma. Methods in Molecular Biology, vol 2265. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1205-7_45
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1205-7_45
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1204-0
Online ISBN: 978-1-0716-1205-7
eBook Packages: Springer Protocols