T-Cell Receptor Gene–Modified T Cells with Shared Renal Cell Carcinoma Specificity for Adoptive T-Cell Therapy

Abstract

Purpose: Adoptive therapy with genetically engineered T cells carrying redirected antigen specificity is a new option for the treatment of cancer. This approach is not yet available for metastatic renal cell carcinoma (RCC), due to the scarcity of therapeutically useful reagents. We analyzed tumor-infiltrating lymphocytes (TIL) from RCC to identify T-cell specificities with shared tumor-specific recognition to develop T-cell receptor (TCR)-engineered T lymphocytes for adoptive therapy of RCC.Experimental Design: We established a T-cell clone from TIL that recognized a human leukocyte antigen (HLA)-A2-restricted tumor antigen. The TCR α-and β-chain genes were isolated, modified by codon optimization and murinization, and retrovirally transduced into peripheral blood lymphocytes (PBL). A TCR-expressing indicator line (B3Z-TCR53) was established to screen for antigen prevalence in RCC, other malignancies, and normal cell counterparts.Results: TCR53-engineered PBL recapitulated the specificity of the TIL and showed tumor-specific HLA-A2-restricted effector activities (IFN-γ, tumor necrosis factor-α, interleukin-2, macrophage inflammatory protein-1β, cytotoxicity). PBL-TCR53 of healthy donors and RCC patients exhibited similar transduction efficiency, expansion, and polyfunctional profile. Using B3Z-TCR53 cells, 130 tumor and normal cells were screened and shared TCR53 peptide: MHC expression was found in >60% of RCC and 25% of tumor lines of other histology, whereas normal tissue cells were not recognized.Conclusions: To date, TCR53 is the only TCR with shared HLA-A2-restricted recognition of RCC. It fulfills the criteria for utilization in TCR gene therapy and advances T cell-based immunotherapy to patients with RCC and other malignancies expressing the TCR ligand.

Chimeric PD-1:28 Receptor Upgrades Low-Avidity T cells and Restores Effector Function of Tumor-Infiltrating Lymphocytes for Adoptive Cell Therapy

“…Constructs, expression vectors, electroporation, and retroviral transduction pMP71 plasmids encoding TCR-T58, TCR-D115 (both tyrosinase-epitope(AA 366-377 )-specific/HLA-A2-restricted), and TCR53 have been described previously (27). Various costimulatory chimeric receptors were designed on the basis of the extracellular domain (ECD) of PD-1 and the intracellular domain (ICD) of CD28 containing the signaling motifs YMNM, PRRP, and PYAP (Fig.…”

Tumor lactic acidosis suppresses CTL function by inhibition of p38 and JNK/c‐Jun activation

“…18 KT195 is a HLA-A2 negative renal cell carcinoma line (tumor cell collection, Deutsches Krebsforschungszentrum, Heidelberg, Germany). T cells retrovirally transduced with the RCC-reactive HLA-A2 restricted T cell receptor (TCR53) are described in Leisegang et al 19 The cell population was frozen until use. One day before the experiment, cells were thawed and rested overnight in CTL medium (M) with 50 U/ml IL-2.…”

“…The same results were observed for TNFa and IL2 (data not shown). Lytic granule exocytosis was also impaired, albeit less severely, with on average 50% 19 was analyzed in the presence of lactic acid. The observed 90% suppression of IFNc production and 40% reduction of degranulation were comparable to that seen for CTL-JB4 (data not shown).…”

High DGK-α and Disabled MAPK Pathways Cause Dysfunction of Human Tumor-Infiltrating CD8+ T Cells That Is Reversible by Pharmacologic Intervention

“…Therefore, the TILs analyzed will exhibit different HLA restrictions and will have different Ag specificity. In the specific situation of RCC, tumor-associated Ags shared among a high percentage of patients are not known (7), precluding the use of one selected pMHC for stimulation if TILs of a larger patient group were to be analyzed. Primary tumor cell cultures corresponding to the respective patient's TILs are not an appropriate source for stimulation because their MHC and/ or Ag presentation may be variable (including loss thereof), and inhibitory ligands may be present (19).…”

“…Clinical evidence includes the observations of spontaneous remission and response to immunotherapy with long-lasting regression in some patients (2,3). Histologically, RCC has a large immune cell infiltrate of CD8 + T lymphocytes with some expressing TCRs that enable the T cells to recognize and kill tumor cells in vitro (4)(5)(6)(7). Despite the local presence of putative tumor-reactive cytotoxic T cells, RCCs are generally not rejected indicating deficits in the cytotoxic response at the tumor site.…”