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Ex Vivo Characterization of Allo-MHC-Restricted T Cells Specific for a Single MHC-Peptide Complex¹

Mikaël J. Pittet^2,*, Asma Gati, Frederic-Anne Le Gal, Gilles Bioley^*, Philippe Guillaume, Magda de Smedt^¶, Jean Plum^¶, Daniel E. Speiser^*, Jean-Charles Cerottini, Pierre-Yves Dietrich, Pedro Romero^3,* and Alfred Zippelius^3,

^* Division of Clinical Onco-Immunology, Ludwig Institute for Cancer Research, University Hospital, Lausanne, Switzerland; Department of Oncology, University Hospital Zurich, Zurich, Switzerland; Division of Oncology, Laboratory of Tumor Immunology, University Hospital, Geneva, Switzerland; Ludwig Institute for Cancer Research, Lausanne Branch, University of Lausanne, Epalinges, Switzerland; and ^¶ Department of Clinical Chemistry, Microbiology and Immunology, University Hospital, Ghent, Belgium

Alloreactive T cells are thought to be a potentially rich source of high-avidity T cells with therapeutic potential since tolerance to self-Ags is restricted to self-MHC recognition. Given the particularly high frequency of alloreactive T cells in the peripheral immune system, we used numerous MHC class I multimers to directly visualize and isolate viral and tumor Ag-specific alloreactive CD8 T cells. In fact, all but one specificities screened were undetectable in ex vivo labeling. In this study, we report the occurrence of CD8 T cells specifically labeled with allo-HLA-A*0201/Melan-A/MART-1_26–35 multimers at frequencies that are in the range of 10^–4 CD8 T cells and are thus detectable ex vivo by flow cytometry. We report the thymic generation and shaping of tumor Ag-specific, alloreactive T cells as well as their fate once seeded in the periphery. We show that these cells resemble their counterparts in HLA-A*0201-positive individuals, based on their structural and functional attributes.

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