MHC-related protein 1–restricted recognition of cancer via a semi-invariant TCR-α chain
Garry Dolton, Hannah Thomas, Li Rong Tan, Cristina Rius Rafael, Stephanie Doetsch, Giulia-Andreea Ionescu, Lucia F. Cardo, Michael D. Crowther, Enas Behiry, Théo Morin, Marine E. Caillaud, Devinder Srai, Lucia Parolini, Md Samiul Hasan, Anna Fuller, Katie Topley, Aaron Wall, Jade R. Hopkins, Nader Omidvar, Caroline Alvares, Joanna Zabkiewicz, John Frater, Barbara Szomolay, Andrew K. Sewell
Garry Dolton, Hannah Thomas, Li Rong Tan, Cristina Rius Rafael, Stephanie Doetsch, Giulia-Andreea Ionescu, Lucia F. Cardo, Michael D. Crowther, Enas Behiry, Théo Morin, Marine E. Caillaud, Devinder Srai, Lucia Parolini, Md Samiul Hasan, Anna Fuller, Katie Topley, Aaron Wall, Jade R. Hopkins, Nader Omidvar, Caroline Alvares, Joanna Zabkiewicz, John Frater, Barbara Szomolay, Andrew K. Sewell
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Research Article Immunology Oncology

MHC-related protein 1–restricted recognition of cancer via a semi-invariant TCR-α chain

Abstract

The T cell antigen presentation platform MR1 consists of 6 allomorphs in humans that differ by no more than 5 amino acids. The principal function of this highly conserved molecule involves presenting microbial metabolites to the abundant mucosal-associated invariant T (MAIT) cell subset. Recent developments suggest that the role of MR1 extends to presenting antigens from cancer cells, a function dependent on the K43 residue in the MR1 antigen binding cleft. Here, we successfully cultured cancer-activated, MR1-restricted T cells from multiple donors and confirmed that they recognized a wide range of cancer types expressing the most common MR1*01 and/or MR1*02 allomorphs (over 95% of the population), while remaining inert to healthy cells including healthy B cells and monocytes. Curiously, in all but one donor these T cells were found to incorporate a conserved TCR-α chain motif, CAXYGGSQGNLIF (where X represents 3–5 amino acids), because of pairing between 10 different TRAV genes and the TRAJ42 gene segment. This semi-invariance in the TCR-α chain is reminiscent of MAIT cells and suggests recognition of a conserved antigen bound to K43.

Authors

Garry Dolton, Hannah Thomas, Li Rong Tan, Cristina Rius Rafael, Stephanie Doetsch, Giulia-Andreea Ionescu, Lucia F. Cardo, Michael D. Crowther, Enas Behiry, Théo Morin, Marine E. Caillaud, Devinder Srai, Lucia Parolini, Md Samiul Hasan, Anna Fuller, Katie Topley, Aaron Wall, Jade R. Hopkins, Nader Omidvar, Caroline Alvares, Joanna Zabkiewicz, John Frater, Barbara Szomolay, Andrew K. Sewell

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Figure 7

Cancer reactivity requires TCR replacement.

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Cancer reactivity requires TCR replacement. (A) WT or TCR-KO E6.1 Jurkat...
(A) WT or TCR-KO E6.1 Jurkat cells with MC.7.G5 or K8T-2 TCRs stained with TRBV25 or TRBV28 antibodies, respectively. Conditions in triplicate, error bars depict SD. (B) WT or TCR-KO E6.1 Jurkat cells with MC.7.G5, MC.27.759S, K8T-1, or K8T-2 TCRs in a CD69 assay (24 hours) with C1R WT or overexpressed MR1*01. Performed over 3 assays: MC.7.G5, MC.27.759S, and K8T-1/K8T-2. Conditions in triplicate, error bars depict SD. Statistics for WT and TCR-KO Jurkat cells versus C1R + MR1*01. Individual P values for a Shapiro-Wilk normality and paired 2-tailed t test. Collective P value for a Wilcoxon signed-rank test. (C) Purified CD8+ T cells from 2 healthy donors (216T and 192D) transduced with MC.7.G5 TCR, comparing KI with TCR replacement. Staining with rCD2 (TCR comarker) and TRBV25 for MC.7.G5 TCR expression. (D) Donors 216T and 192D CD8+ T cells, either untransduced, MC.7.G5 TCR KI or TCR replaced in an overnight activation assays with cancer cells, followed by TNF ELISA. MC.7.G5 clone, included for comparison. Duplicate conditions. (E) Donor 216T CD8+ T cells, either untransduced, MC.7.G5 TCR KI or TCR replaced in an overnight activation assay with cancer cells, followed by Granzyme B ELISA. Triplicate conditions, error bars depict SD. (F) Donor 216T CD8+ T cells, either untransduced, MC.7.G5 TCR KI or TCR replaced in an overnight activation assay versus MR1*01 or MR1*01/*02 cancer cells, followed by TNF ELISA. Triplicate conditions, with error bars depicting SD. P value for a multivariate permutation test for paired comparison. (G) Donor 216T CD8+ T cells, either untransduced, MC.7.G5 TCR KI or TCR replaced in an overnight flow cytometry–based killing assay at a 1:1 ratio with cancer cells FM72, or PBMCs from 3 healthy donors. CD14 and CD19 antibodies used to identify monocytes (M) and B cells (B) during analysis. MC.7.G5 clone data repeated (*) on each graph for comparison. Cancer cells include melanoma (FM72, MEL624, MEL526, FM3, FM88, and FM74), kidney (ACHN), pancreatic (BxPC-3 and MIA PaCa-2), leukemia/CML (K-562), breast (MCF-7), ovarian (A2780), and cervical (SiHa) cancers.