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Expansion of human cytomegalovirus‐specific T lymphocytes from unfractionated peripheral blood mononuclear cells with artificial antigen‐presenting cells
Author(s) -
Paine Ananta,
Oelke Mathias,
Blasczyk Rainer,
EizVesper Britta
Publication year - 2007
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.1537-2995.2007.01439.x
Subject(s) - cytotoxic t cell , peripheral blood mononuclear cell , immunology , cd8 , antigen , human leukocyte antigen , biology , microbiology and biotechnology , in vitro , biochemistry
BACKGROUND: The aim of this study was to find a simple and feasible method for ex vivo expansion of human cytomegalovirus (CMV)‐specific cytotoxic T cells from unfractionated peripheral blood mononuclear cells (PBMNCs). STUDY DESIGN AND METHODS: Unfractionated PBMNCs from three HLA‐A*0201‐CMV–seropositive donors were stimulated with CMVpp65 495‐503 peptide–loaded HLA‐A*0201‐immunoglobulin fusion protein (HLA‐A2‐Ig) based artificial antigen‐presenting cells (aAPCs) on Day 1. Once a week the CMV‐specific T cells were harvested and restimulated with fresh aAPCs. T‐cell cultures were maintained for 28 days and then analyzed. RESULTS: With aAPCs and starting with 1 × 10 7 freshly isolated PBMNCs that were less than 0.1 percent CMV‐specific, more than 1 × 10 7 T cells with a CMV‐specific frequency greater than 93 percent in all donors tested were generated. Expanded CD8+ cytotoxic T lymphocytes were functionally active and showed antigen‐specific secretion of interferon‐γ and cytotoxic activity. No alloreactivity against unpulsed HLA‐A*0201–positive cells was detected. CONCLUSION: Herein is reported the successful in vitro expansion of CMV‐specific cytotoxic CD8+ T cells from unfractionated PBMNCs of healthy CMV‐seropositive blood donors by the use of HLA‐A2‐Ig–based aAPCs. This study demonstrates that more than 1 × 10 7 CMV‐specific T cells can be generated from approximately 1 × 10 7 unfractionated PBMNCs within 1 month under highly reproducible conditions.