Isolation and Characterization of an HLA-DRB1*04-Restricted HPV16-E7 T Cell Receptor for Cancer Immunotherapy

Abstract

High-risk human papillomavirus (HPV) infection is a causal factor in oropharyngeal and gynecological malignancies, and development of HPV-targeted immunotherapy could be used to treat patients with these cancers. T cell-mediated adoptive immunotherapy targeting E6 and E7, two HPV16 proteins consistently expressed in tumor cells, appears to be both attractive and safe. However, isolation of HPV-specific T cells is difficult owing to the low frequency of these cell precursors in the peripheral blood. In addition, HPV-positive cancer cells often down-regulate major histocompatibility complex (MHC) class I expression ex vivo, limiting the efficacy of MHC class I-restricted approaches. Of particular interest is that both CD4 and CD8 T cells can mediate the responses. Given that CD4 T cells play a critical role in coordinating effective antitumor responses, the generation of a T helper response in patients with HPV16-associated malignancies would unleash the ultimate potential of immunotherapy. In this view, T-cell receptor (TCR) gene transfer could be a relevant strategy to generate HPV16-E7-specific and MHC class II-restricted T cells in sufficient numbers. An HPV16-E7/HLA-DRB1*04 TCR has been isolated from a cancer patient with complete response, and retroviral particles encoding this TCR have been produced. The transgenic TCR is highly expressed in transduced T cells, with a functional inducible caspase-9 suicide gene safety cassette. TCR transgenic T cells are HPV16-E7 specific and HLA-DRB1*04 restricted, as determined by interferon (IFN)-γ secretion. CD8 and CD4 T cells are equivalently transduced and secrete interleukin-2 and IFN-γ when cultured with appropriate targets. We also demonstrate that TCR transgenic T cells recognize the endogenously processed and presented HPV16-E7 peptide. In conclusion, our data indicate that the production of MHC class II-restricted HPV16-E7-specific T cells is feasible through TCR gene transfer and could be used for immunotherapy.

The Tumor Microenvironment and Immunotherapy of Oropharyngeal Squamous Cell Carcinoma

“…Another study reported the isolation of an HLA-DRB1*04 restricted TCR reactive to the HPV16 E7 70-89 epitope. TCRtransduced cells were shown to specifically produce IFNg when stimulated with HPV16 E6 and E7 containing tumor lysate (160). The isolation of TCR responsive to HPV epitopes restricted by a series of HLA class I and II molecules will lead to a warehouse of HPV-specific TCR required for personalized treatment of patients with recurrent or metastatic HPV-induced OPSCC.…”

HSV-2-Specific Human Female Reproductive Tract Tissue Resident Memory T Cells Recognize Diverse HSV Antigens

“…Figure 1 is an example in which stimulation of PBMC with moDC-mock-HeLa led to non-zero, CD137-expressing CD4 and CD8 T cells, consistent with our previous publication ( 18 ). HPV-specific T cells are rare: in vitro elicitation of HPV E6 or E7-reactive T cells from PBMC uses repetitive cycles of re-stimulation with high concentrations of peptides ( 109 ), and in vivo vaccinations use many cycles of peptide-loaded DC ( 110 ). It would be possible to query our HSV-reactive T cell lines with HPV expression constructs ( 111 ) to detect HPV-specific reactivity in our system.…”

TCR-engineered T cell therapy in solid tumors: State of the art and perspectives

“…( 52 – 54 ). More recently, some groups combined VDJ sequencing and reference sequences from the ImMunoGeneTics database to identify functional TCRs ( 55 , 56 ). The development of single-cell RNA sequencing (scRNA-seq) focused on TCR sequences (scTCR-seq) has also facilitated the identification of TCRs from primed T cells.…”