Isolation and characterization of NY-ESO-1–specific T cell receptors restricted on various MHC molecules

Abstract

SignificanceT immune cells can be engineered to express tumor-specific T cell receptor (TCR) genes and thereby kill cancer cells. This approach—termed TCR gene therapy—is effective but can cause serious adverse events if the target is also expressed in healthy, noncancerous tissue. NY-ESO-1 is a tumor-specific antigen that has been targeted successfully and safely through TCR gene therapies for melanoma, synovial sarcoma, and myeloma. However, trials to date have focused exclusively on a single NY-ESO-1–derived epitope presented on HLA-A*02:01, limiting application to patients expressing that allele. In this work, we isolate TCRs that collectively recognize multiple NY-ESO-1–derived epitopes presented by multiple MHC alleles. We thereby outline a general approach for expanding targeted immunotherapies to more diverse MHC haplotypes.

High throughput pMHC-I tetramer library production using chaperone-mediated peptide exchange

“…7d). The observed diversity in TCR CDR3 regions that recognize NY-ESO-1 tetramers are consistent with the known high cross-reactivity of this epitope 33 , and provide clues for engineering more exclusive TCRs for immunotherapy applications 34 (Fig. 7e).…”

Harnessing neoantigen specific CD4 T cells for cancer immunotherapy

“…Whereas the majority of early studies have prioritized single TCRs recognizing targets in the context of the common HLA-A*02:01 allele (of North American and European Caucasian populations), only recently have investigators searched for TCRs restricted to other common alleles. 123 Priority targeting of known driver mutations, which are more likely to be clonal "trunk" mutations, will be advantageous so as to limit immune escape from antigen loss or incomplete clearance due to preexisting intratumor heterogeneity. [124][125][126] A final consideration for effective NeoAg-specific therapies is that of optimal TCR avidity.…”

Targeting monoamine oxidase A-regulated TAM polarization for cancer immunotherapy

“…The HEK 293T and Phoenix-ECO retroviral packaging cell lines, the A375 human melanoma cell line, and the L-929 mouse connective tissue cell line were purchased from the American Type Culture Collection (ATCC). The A375-A2-ESO cell line was previously reported 98 .…”

“…The Retro/ESO-TCR vector was constructed by inserting into the parental pMSGV vector a synthetic gene encoding an HLA-A2-restricted, NY-ESO-1 tumour antigen-speci c human CD8 TCR (clone 3A1) 98 . Vsv-gpseudotyped Retro/ESO-TCR retroviruses were generated by transfecting HEK 293T cells following a standard calcium precipitation protocol and an ultracentrifugation concentration protocol 102 ; the viruses were then used to transduce PG13 cells to generate a stable retroviral packaging cell line producing GALV-pseudotyped Retro/ESO-TCR retroviruses (denoted as the PG13-ESO-TCR cell line).…”

“…Healthy donor PBMCs were cultured in a 12-well plate in C10 medium (1 x 10 6 cells/ml/well) for 2 days, stimulated with Dynabeads™ Human T-Activator CD3/CD28 (10 μl/ml) (GIBCO, 11161D) and recombinant human IL-2 (20 ng/ml) (Peprotech). After 2 days, dynabeads were removed and cells were spin-infected with frozen-thawed Retro/ESO-TCR retroviral supernatants supplemented with polybrene (10 µg/ml) at 660 g at 30 °C for 90 minutes following an established protocol 98 . Transduced human CD8 + T cells (denoted as ESO-T cells) were expanded for another 6-8 days in C10 medium containing recombinant human IL-2 (20 ng/ml) (Peprotech), and then cryopreserved for future use.…”