Abstract
Killer T-cells play important roles in immunity to infection and cancer by detecting intracellular anomalies at the cell surface and destroying the cells that bear them. Conventional killer T-cells scan the intracellular proteome by sampling peptides presented at the cell surface by major histocompatibility complex (MHC) molecules. It is becoming apparent that some T-cells can also respond to pathogens and neoplasms by sensing intracellular changes through molecules other than MHC. We describe an unbiased methodology for T-cell receptor ligand discovery that requires no a priori knowledge regarding the nature of the antigen.
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
Porter DL, Levine BL et al (2011) Chimeric antigen receptor–modified T cells in chronic lymphoid leukemia. N Engl J Med 356:725–733
Grupp SA, Kalos M, Barrett D et al (2013) Chimeric antigen receptor–modified T cells for acute lymphoid leukemia. N Engl J Med 368:1509–1518
Andersen R, Donia M, Ellebaek E et al (2016) Long-lasting complete responses in patients with metastatic melanoma after adoptive cell therapy with tumor-infiltrating lymphocytes and an attenuated il2 regimen. Clin Cancer Res 22:3734–3745
Kenter G, Mezzadra R, Slagter M et al (2018) Low and variable tumor reactivity of the intratumoral TCR repertoire in human cancers. Nat Med 25:89–94
Schmitt TM, Ragnarsson GB, Greenberg PD (2009) T cell receptor gene therapy for cancer. Hum Gene Ther 20:1240–1248
Legut M, Dolton G, Mian AA et al (2018) CRISPR-mediated TCR replacement generates superior anticancer transgenic t cells. Blood 131:311–322
Tan MP, Gerry AB, Brewer JE et al (2015) T cell receptor binding affinity governs the functional profile of cancer-specific CD8 + T cells. Clin Exp Immunol 180:255–270
Bridgeman JS, Sewell AK, Miles JJ et al (2012) Structural and biophysical determinants of αβ T-cell antigen recognition. Immunology 135:9–18
Gonzalez-Galarza FF, McCabe A, Dos Santos EJM et al (2020) Allele frequency net database (AFND) 2020 update: gold-standard data classification, open access genotype data and new query tools. Nucleic Acids Res 48:D783–D788
Attaf M, Legut M, Cole DK et al (2015) The T cell antigen receptor: the Swiss army knife of the immune system. Clin Exp Immunol 181:1–18
Crowther MD, Dolton G, Legut M et al (2020) Genome-wide CRISPR-Cas9 screening reveals ubiquitous T cell cancer targeting via the monomorphic MHC class I-related protein MR1. Nat Immunol. 21(2):178–185
Crowther MD, Sewell AK (2021) The burgeoning role of MR1-restricted T-cells in infection, cancer and autoimmune disease. Curr Opin Immunol. https://doi.org/10.1016/j.coi.2020.12.002
Kjer-Nielsen L, Patel O, Corbett AJ et al (2012) MR1 presents microbial vitamin B metabolites to MAIT cells. Nature 491:717–723
Tilloy F, Treiner E, Park SH et al (1999) An invariant T cell receptor alpha chain defines a novel TAP-independent major histocompatibility complex class Ib-restricted alpha/beta T cell subpopulation in mammals. J Exp Med 189:1907–1921
Morita CT, Beckman EM, Bukowski JF et al (1995) Direct presentation of nonpeptide Prenyl pyrophosphate antigens to human gamma delta T cells. Immunity 3:495–507
Karunakaran MM, Willcox CR, Salim M et al (2020) Butyrophilin-2A1 directly binds germline-encoded regions of the Vγ9Vδ2 TCR and is essential for Phosphoantigen sensing. Immunity 52:487–498.e6
Doench JG, Fusi N, Sullender M et al (2016) Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. Nat Biotechnol 34:184–191
Wang B, Wang M, Zhang W et al (2019) Integrative analysis of pooled CRISPR genetic screens using MAGeCKFlute. Nat Protoc 14:756–780
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Crowther, M.D., Legut, M., Sewell, A.K. (2022). Ligand Identification for Orphan MHC-Agnostic T-Cell Receptors by Whole Genome CRISPR–Cas9 Screening. In: Huang, H., Davis, M.M. (eds) T-Cell Repertoire Characterization. Methods in Molecular Biology, vol 2574. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2712-9_1
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2712-9_1
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2711-2
Online ISBN: 978-1-0716-2712-9
eBook Packages: Springer Protocols