TCR γ4δ1-Engineered αβT Cells Exhibit Effective Antitumor Activity

Kangxia He,Jianmin Zhang,Wei He,Lianxian Cui,Hongqin You,Yuxia Li

doi:10.2119/molmed.2016.00023

Abstract

T cell engineering with T cell receptors (TCRs) specific for tumors plays an important role in adoptive T-cell transfer (ATC) therapy for cancer. Here, we present a novel strategy to redirect peripheral blood-derived αβT cells against tumors via TCRγ4δ1 gene transduction. The broad-spectrum anti-tumor activity of TCRδ1 cells in innate immunity is dependent on CDR3δ1. TCRγ4δ1-engineered αβT cells were prepared by lentiviral transduction and characterized by analyzing in vitro and in vivo cytotoxicity to tumors, ability of proliferation and cytokine production, and their potential role in autoimmunity. Results show TCRγ4δ1 genes were transduced to approximately 36% of polyclonal αβT cells. TCRγ4δ1-engineered αβT cells exhibited an effective in-vitro TCRγδ-dependent cytotoxicity against various tumor cells via the perforin-granzyme pathway. They also showed a strong proliferative capacity and robust cytokine production. TCRγ4δ1-engineered αβT cells neither expressed mixed TCR dimers nor bound/killed normal cells in vitro. More importantly, adoptive transfer of TCRγ4δ1-engineered αβT cells into nude mice bearing a human HepG2 cell line significantly suppressed tumor growth. Our results demonstrate a novel role for TCRγ4δ1 in gene therapy and ATC for cancer.

Highlights

Adoptive T cell transfer (ACT) is a promising immunotherapy strategy for cancer
We previously identified a highfrequency complementary determining region 3 (CDR3) dominant sequence (CDR3δ1: CAFLPHADKLIFGKG), termed GTM, in TCRδ1 chain from tumor infiltrating lymphocytes (TILs) in human gastric cancer through RT-PCR and analysis of a large number of CDR3δ1 sequences
We confirmed that the CDR3δ1 peptide played a crucial role in tumor antigen recognition and bound to a wide variety of tumor cell lines and tissues similar to intact TCRγ4δ1

Summary

Introduction

Adoptive T cell transfer (ACT) is a promising immunotherapy strategy for cancer. Chimeric antigen receptor (CAR)-modified T cells have shown promise in the treatment of B cell malignancy. [1] ACT immunotherapy for solid tumors faces the challenge of specificity when targeting tumors. Vδ1 T cells are common in mucosa, especially the submucosal areas of the gastrointestinal, respiratory and genitourinary tracts. They recognize MHC class I–related molecules A and B (MICA and MICB) and UL-16–binding proteins (ULBPs) expressed at variable levels on epithelial tumor cells and some leukemias and lymphomas. [8] Due to broad-spectrum tumor recognition of TCRγδ, TCRgd gene transduction into effector T cells, such as αβT cells, may be an attractive therapeutic approach. It appears to resolve the fundamental problem of tumor targeting not found in TCRαβ. Previous studies by other groups and our laboratory have confirmed that TCRg 9d 2-transduced αβT cells, or TCRg 9d 2-modified peripheral blood mononuclear cells (PBMCs), mediate cytotoxicity against a broad range of tumor cell lines in vitro and suppress tumor growth in Daudi or SKOV3 tumor cell–bearing mice models. [9,10]

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Results

Conclusion