MHC-restricted fratricide of human lymphocytes expressing survivin-specific transgenic T cell receptors
Matthias Leisegang, Susanne Wilde, Stefani Spranger, Slavoljub Milosevic, Bernhard Frankenberger, Wolfgang Uckert, Dolores J. Schendel
Matthias Leisegang, Susanne Wilde, Stefani Spranger, Slavoljub Milosevic, Bernhard Frankenberger, Wolfgang Uckert, Dolores J. Schendel
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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.