MHC-restricted fratricide of human lymphocytes expressing survivin-specific transgenic T cell receptors
Matthias Leisegang, … , Wolfgang Uckert, Dolores J. Schendel
Matthias Leisegang, … , Wolfgang Uckert, Dolores J. Schendel
Published October 11, 2010
Citation Information: J Clin Invest. 2010;120(11):3869-3877. https://doi.org/10.1172/JCI43437.
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Research Article Oncology

MHC-restricted fratricide of human lymphocytes expressing survivin-specific transgenic T cell receptors

Abstract

The apoptosis inhibitor protein survivin is overexpressed in many tumors, making it a candidate target molecule for various forms of immunotherapy. To explore survivin as a target antigen for adoptive T cell therapy using lymphocytes expressing survivin-specific transgenic T cell receptors (Tg-TCRs), we isolated HLA-A2–allorestricted survivin-specific T cells with high functional avidity. Lymphocytes expressing Tg-TCRs were derived from these T cells and specifically recognized HLA-A2+ survivin+ tumor cells. Surprisingly, HLA-A2+ but not HLA-A2– lymphocytes expressing Tg-TCRs underwent extensive apoptosis over time. This demise was caused by HLA-A2–restricted fratricide that occurred due to survivin expression in lymphocytes, which created ligands for Tg-TCR recognition. Therefore, survivin-specific TCR gene therapy would be limited to application in HLA-A2–mismatched stem cell transplantation. We also noted that lymphocytes that expressed survivin-specific Tg-TCRs killed T cell clones of various specificities derived from HLA-A2+ but not HLA-A2– donors. These results raise a general question regarding the development of cancer vaccines that target proteins that are also expressed in activated lymphocytes, since induction of high-avidity T cells that expand in lymph nodes following vaccination or later accumulate at tumor sites might limit themselves by self-MHC–restricted fratricide while at the same time inadvertently eliminating neighboring T cells of other specificities.

Authors

Matthias Leisegang, Susanne Wilde, Stefani Spranger, Slavoljub Milosevic, Bernhard Frankenberger, Wolfgang Uckert, Dolores J. Schendel

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Figure 2

Redirection of antigen specificity by retroviral transfer of survivin-specific TCR genes.

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Redirection of antigen specificity by retroviral transfer of survivin-sp...
PBLs of a healthy HLA-A2– donors were transduced with TCR-A71, -A66, and -A72. (A) Unsorted TCR-transduced PBLs were analyzed on day 10 for Tg-TCR expression using murine TCRβ constant region–specific antibody. (B) The relative cytotoxicity is given for nonsorted TCR-transduced PBLs following stimulation with T2 cells loaded with graded amounts of survivin peptide at an E/T ratio of 20:1. Relative values of half-maximal killing are depicted at the right. (C) RNA expression of survivin in tumor cells was assessed by RT-PCR using survivin-specific primers and β2-microglobulin–specific primers (β2m) as a control. Survivin-specific cytotoxicity of TCR-modified PBLs was assessed in a standard 4-hour chromium release assay using different tumor cell lines at varying E/T ratios: (D) UT-SCC-15, U-373, and FM-86 (all: A2+, S+) and (E) KT-195, KT-195-VC (A2–, S+), KT-195-A2 (A2+, S+) as target cells. Untransduced PBLs and PBLs transduced with a GFP control vector served as controls. Cytotoxicity data represent means of duplicates measured at each E/T ratio or peptide concentration. (F) IFN-γ release at 24 hours is depicted following coculture with tumor cell lines at an E/T ratio of 2:1. (G) Unloaded or survivin or flu peptide–loaded aAPCs were cocultured with either untransduced PBLs or PBLs expressing TCR-A72 at an E/T ratio of 1:2. IFN-γ values for F and G are shown as mean of duplicates ± mean deviation. These experiments were done with 6 different donors, except for B and G (n = 2). Asterisk indicates values below the detection limit.