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Ex vivo generation of human cytomegalovirus‐specific cytotoxic T cells by peptide‐pulsed dendritic cells
Author(s) -
Kleihauer Annette,
Grigoleit Ulrich,
Hebart Holger,
Moris Arnaud,
Brossart Peter,
Muhm Alexandra,
Stevanovic Stefan,
Rammensee Hans Georg,
Sinzger Christian,
Riegler Susanne,
Jahn Gerhard,
Kanz Lothar,
Einsele Hermann
Publication year - 2001
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.2001.02681.x
Subject(s) - cytotoxic t cell , human cytomegalovirus , ex vivo , adoptive cell transfer , cd8 , transplantation , immunotherapy , t cell , biology , stem cell , immunology , virology , antigen , immune system , in vivo , medicine , in vitro , microbiology and biotechnology , virus , biochemistry
Adoptive transfer of donor‐derived human cytomegalovirus (HCMV)‐specific T‐cell clones can restore protective immunity after stem cell transplantation. Ex vivo induction of HCMV‐specific T cells using HCMV‐infected fibroblasts as stimulator cells confines this approach to HCMV‐seropositive donors and requires the presence of infectious virus during the stimulation procedure. In this study, we describe a potential alternative strategy to generate HCMV‐specific T cells ex vivo for adoptive immunotherapy. Generation of HCMV‐specific cytotoxic T lymphocytes (CTLs) ex vivo was investigated using peptide‐pulsed dendritic cells as antigen‐presenting cells. HCMV‐specific T cells were generated and sufficiently expanded for adoptive immunotherapy in 6 out of 14 HCMV‐seropositive and 2 out of 11 HCMV‐seronegative donors. The CTLs recognized HCMV‐infected autologous fibroblasts. No lysis was observed with either non‐infected autologous or HLA‐mismatched infected fibroblasts. Staining with tetrameric HLA/peptide complexes revealed significant enrichment for peptide‐specific T cells of up to 28% and > 90% of CD8 + T cells after three and five specific stimulations respectively. In addition, the expansion rates indicated that ex vivo generation of > 1 × 10 9 HCMV‐specific T cells was possible after 6–7 weeks when cultures were initiated with 1–5 × 10 6 responder cells. Thus, the approach with peptide‐pulsed DCs to generate HCMV‐specific CTLs is feasible for clinical application after allogeneic stem cell transplantation.