1. Lysosome-associated membrane glycoprotein 1 predicts fratricide amongst T cell receptor transgenic CD8+ T cells directed against tumor-associated antigens
- Author
-
Julia S. Gerke, Melanie Thiede, Stefan Burdach, Franziska Blaeschke, Michaela C. Baldauf, Gunther Richter, Thomas G. P. Grunewald, Andreas Kirschner, Uwe Thiel, and Dirk H. Busch
- Subjects
0301 basic medicine ,Adoptive cell transfer ,Genotype ,Annexins ,Colon ,T cell ,Receptors, Antigen, T-Cell ,TCR transgenic T cells ,Cell Separation ,CD8-Positive T-Lymphocytes ,Immunotherapy, Adoptive ,Retina ,03 medical and health sciences ,0302 clinical medicine ,CD107a ,Antigens, Neoplasm ,Lysosomal-Associated Membrane Protein 1 ,HLA-A2 Antigen ,Medicine ,Cytotoxic T cell ,Humans ,Transgenes ,Adaptor Proteins, Signal Transducing ,Oligonucleotide Array Sequence Analysis ,business.industry ,ELISPOT ,T-cell receptor ,amino-acid exchange scan ,Lysosome-Associated Membrane Glycoproteins ,Flow Cytometry ,Molecular biology ,Adoptive Transfer ,Tumor antigen ,ddc ,Transplantation ,030104 developmental biology ,medicine.anatomical_structure ,Retroviridae ,Oncology ,030220 oncology & carcinogenesis ,cross reactivity ,Receptors, Adrenergic, beta-3 ,Immunology ,Leukocytes, Mononuclear ,business ,K562 Cells ,fratricide ,CD8 ,Research Paper - Abstract
// Andreas Kirschner 1 , Melanie Thiede 1 , Franziska Blaeschke 1, 2 , Gunther H.S. Richter 1 , Julia S. Gerke 3 , Michaela C. Baldauf 3 , Thomas G.P. Grunewald 3, 4, 5 , Dirk H. Busch 6 , Stefan Burdach 1, * , Uwe Thiel 1, * 1 Laboratory for Functional Genomics and Transplantation Biology, Departments of Pediatrics and Children’s Cancer Research Center, Klinikum rechts der Isar, Technische Universitat Munchen, Munich, Germany 2 Laboratory for Immunotherapy, Dr. von Hauner Children’s Hospital, Medical center of the LMU Munich, Munich, Germany 3 Laboratory for Pediatric Sarcoma Biology, Institute of Pathology of the LMU Munich, Munich, Germany 4 German Cancer Consortium (DKTK), Heidelberg, Germany 5 German Cancer Research Center (DKFZ), Heidelberg, Germany 6 Institute for Medical Microbiology, Immunology and Hygiene, Technische Universitat Munchen, Munich, Germany * Share senior authorship Correspondence to: Andreas Kirschner, email: andi.kirschner@tum.de Keywords: fratricide, CD107a, TCR transgenic T cells, cross reactivity, amino-acid exchange scan Received: April 18, 2016 Accepted: May 30, 2016 Published: July 18, 2016 ABSTRACT Aim: Autologous as well as allogeneic CD8 + T cells transduced with tumor antigen specific T cell receptors (TCR) may cause significant tumor lysis upon adoptive transfer. Besides unpredictable life-threatening off-target effects, these TCRs may unexpectedly commit fratricide. We hypothesized lysosome-associated membrane glycoprotein 1 (LAMP1, CD107a) to be a marker for fratricide in TCR transgenic CD8 + T cells. Methods: We identified HLA-A*02:01/peptide-restricted T cells directed against ADRB3 295 . After TCR identification, we generated HLA-A*02:01/peptide restricted TCR transgenic T cells by retroviral transduction and tested T cell expansion rates as well as A*02:01/peptide recognition and ES killing in ELISpot and xCELLigence assays. Expansion arrest was analyzed via Annexin and CD107a staining. Results were compared to CHM1 319 -TCR transgenic T cells. Results: Beta-3-adrenergic receptor (ADRB3) as well as chondromodulin-1 (CHM1) are over-expressed in Ewing Sarcoma (ES) but not on T cells. TCR transgenic T cells demonstrated HLA-A*02:01/ADRB3 295 mediated ES recognition and killing in ELISpot and xCELLigence assays. 24h after TCR transduction, CD107a expression correlated with low expansion rates due to apoptosis of ADRB3 specific T cells in contrast to CHM1 specific transgenic T cells. Amino-acid exchange scans clearly indicated the cross-reactive potential of HLA-A*02:01/ADRB3 295 - and HLA-A*02:01/CHM1 319 -TCR transgenic T cells. Comparison of peptide motive binding affinities revealed extended fratricide among ADRB3 295 specific TCR transgenic T cells in contrast to CHM1 319 . Conclusion: Amino-acid exchange scans alone predict TCR cross-reactivity with little specificity and thus require additional assessment of potentially cross-reactive HLA-A*02:01 binding candidates. CD107a positivity is a marker for fratricide of CD8 + TCR transgenic T cells.
- Published
- 2016