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Clinical‐scale expansion of human cytomegalovirus–specific cytotoxic T lymphocytes from peripheral blood mononuclear cells requiring single‐peptide stimulation and feeder cells but not additional antigen‐presenting cells
Author(s) -
Sakagawa Hisako,
Azuma Hiroshi,
Fujihara Mitsuhiro,
Ikeda Hisami
Publication year - 2006
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2006.00773.x
Subject(s) - cytotoxic t cell , peripheral blood mononuclear cell , t cell , microbiology and biotechnology , antigen , biology , antigen presenting cell , immunology , population , immune system , medicine , in vitro , biochemistry , environmental health
BACKGROUND: The aim of this study was to find a simple and feasible method for ex vivo expansion of human cytomegalovirus (HCMV)‐specific cytotoxic T cells from peripheral blood mononuclear cells (PBMNCs) without the aid of exogenous antigen‐presenting cells (APCs) such as cultured dendritic cells. STUDY DESIGN AND METHODS: PBMNCs from three HLA‐A*2402‐seropositive donors were stimulated with HCMV pp65 341‐350 peptide on Day 1 and then cultured with interleukin‐2 and allogeneic feeder cells for 3 to 4 weeks. HCMV peptide–specific T cells were purified with HLA‐A*2402/pp65 341‐350 tetramer on Days 12 to 13 and harvested on Days 23 to 27. RESULTS: The initial numbers of PBMNCs were 2 × 10 7 , 1.5 × 10 7 , and 2.5 × 10 7 and the increases in HCMV peptide–specific T cells were 3.5 × 10 4 ‐, 2.0 × 10 3 ‐, and 1.1 × 10 3 ‐fold, respectively. The estimated final numbers of tetramer‐positive cells were 9.1 × 10 7 , 9.0 × 10 6 , and 5.3 × 10 6 , respectively. The purities of the tetramer‐positive cell population in culture were 72.6, 75.0, and 80.9 percent, respectively. The cells killed peptide‐pulsed B‐lympoblastoid cell lines and secreted interferon‐γ in a HLA‐restricted manner. They did not have natural killer cell activity or lymphokine activated killer cell activity. Most of them had an effector‐memory phenotype. They did not express killer inhibitory receptors. CONCLUSION: This method makes it possible to obtain more than 1 × 10 7 HCMV‐specific T cells from approximately 2 × 10 7 to 5 × 10 7 PBMNCs without exogenous APCs such as cultured dendritic cells.