The T cell repertoire selected in vitro against EBV: diversity, specificity, and improved purification through early IL-2 receptor alpha-chain (CD25)-positive selection.
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.
Ibisch
C
C
Institut de Biologie, Institut National de la Santé et de la Recherche Médicale, Nantes, France.
Saulquin
X
X
Gallot
G
G
Vivien
R
R
Ferrand
C
C
Tiberghien
P
P
Houssaint
E
E
Vié
H
H
D000328:Adult; D000818:Animals; D000956:Antigens, Viral / Q000032:analysis / Q000235:genetics / Q000276:immunology; D002461:Cell Line, Transformed; D002469:Cell Separation / Q000379:methods*; D002472:Cell Transformation, Viral; D002999:Clone Cells; D018920:Coculture Techniques; D018984:Epitopes, T-Lymphocyte / Q000096:biosynthesis* / Q000276:immunology; D015332:Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D016130:Immunophenotyping; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000096:biosynthesis* / Q000235:genetics; D015375:Receptors, Interleukin-2 / Q000096:biosynthesis*; D016176:T-Lymphocyte Subsets / Q000166:cytology* / Q000276:immunology / Q000378:metabolism* / Q000821:virology
D000956:Antigens, Viral; D018984:Epitopes, T-Lymphocyte; D016693:Receptors, Antigen, T-Cell, alpha-beta; D015375:Receptors, Interleukin-2