Abstract:
:Based upon the success of using polyclonal, Epstein-Barr virus (EBV)-specific CTL lines for the prophylaxis and treatment of patients with post-transplant lymphoproliferative disease (PTLPD), there is now considerable incentive to develop CTL directed against the sub-dominant EBV antigens EBNA1, LMP1 and LMP2, which are expressed by the tumor cells of Hodgkin disease and nasopharyngeal carcinoma. To develop a system for generating LMP2a-specific CTL in vitro, we transfected autologous immature dendritic cells (DC), which had been grown in the absence of serum, with LMP2a RNA in the presence of the cationic lipid DOTAP. This transfection method did not adversely affect the DC in terms of immunophenotyping and they expressed high levels of HLA class I and II and critical costimulatory molecules. These LMP2a(+) DC, as compared to DC which had been transfected with irrelevant RNA, were shown to be highly immunostimulatory in autologous mixed lymphocyte reactions and, importantly, could stimulate the generation of CD8(+) and CD4(+) CTL which exclusively recognized LMP2a-expressing targets. This specific cytotoxicity was confirmed using antibody blocking experiments and cytotoxicity assays of the separated T cell subsets. Using this DC-based system we could also reactivate LMP2a-specific memory in EBV-seropositive donors whose polyclonal CTL response to LCL stimulation did not contain a LMP2a-specific component.
journal_name
Eur J Immunoljournal_title
European journal of immunologyauthors
Su Z,Peluso MV,Raffegerst SH,Schendel DJ,Roskrow MAdoi
10.1002/1521-4141(200103)31:3<947::aid-immu947>3.0keywords:
subject
Has Abstractpub_date
2001-03-01 00:00:00pages
947-58issue
3eissn
0014-2980issn
1521-4141pii
10.1002/1521-4141(200103)31:3<947::AID-IMMU947>3.0journal_volume
31pub_type
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