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The Journal of Immunology, 2000, 164: 4924-4932.
Copyright © 2000 by The American Association of Immunologists

The T Cell Repertoire Selected In Vitro Against EBV: Diversity, Specificity, and Improved Purification Through Early IL-2 Receptor {alpha}-Chain (CD25)-Positive Selection1

Catherine Ibisch2,*, Xavier Saulquin2,*, Géraldine Gallot*, Régine Vivien*, Christophe Ferrand{dagger}, Pierre Tiberghien{dagger}, Elisabeth Houssaint* and Henri Vié3,*

* Institut de Biologie, Institut National de la Santé et de la Recherche Médicale, Nantes, France; and {dagger} Etablissement de Transfusion Sanguine de Franche Comté, Besançon, France

Polyclonal T cell lines specific for EBV proteins have proved efficient in preventing EBV-related immunoblastic lymphoma after allogeneic bone marrow transplantation. To gain insight into the composition of the EBV-specific T cell repertoire that ensured patient protection, we performed for the first time an extensive characterization of eight cytotoxic T cell lines selected in vitro against EBV-transformed autologous lymphoblastoid cell lines (BLCL). These T cell lines consist of 50–100 distinct T cell clones, of which 32–96% are specific for autologous BLCL. Moreover, we demonstrate that reactivities against only five EBV proteins (BZLF1, BMLF1, EBNA-3A, EBNA-3C, and LMP2) cover 86% (32/37) of the specificities detected. In addition, we describe an improved method of T cell harvesting using a CD25 selection procedure which reduces the time required to obtain specific T cells and improves the purity of EBV-specific T cells, thus showing promise for use in adoptive transfer protocols.




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