Antigen presenting cells transfected with LMP2a RNA induce CD4+ LMP2a-specific cytotoxic T lymphocytes which kill via a Fas-independent mechanism.
Recent reports have demonstrated that EBV can be used as a target of specific CTL-based treatments in severe chronic EBV, immunoblastic B cell lymphoma and Hodgkin's disease (HD). Based upon the promising results form these in vivo studies, it has been suggested that an antigen-specific CTL-based immunotherapy may be of benefit in treating EBV-associated tumors such as HD and nasopharyngeal carcinoma (NPC) which express the potentially immunogenic antigens, LMP1 and LMP2a. Recent work form our group has demonstrated that LMP2a-specific CTLs may be generated in vitro using autologous antigen presenting cells which have been transfected with polyadenylated LMP2a RNA in the presence of a cationic lipid. In this study, we demonstrate that the presence of the lipid enhances dendritic cell (DC) transfection efficiency and appears to protect the intracellular LMP2a RNA from degradation by cellular RNAses. Significantly, these improvements resulted in the transfected DCs having a superior ability to stimulate autologous T cell proliferation. These LMP2a + DCs were used to stimulate LMP2a-specific effector cells which were predominantly a mixture of cytotoxic and helper CD4+ T cells. The molecular mechanisms whereby these CD4+ T cells lyzed their LMP2a-expressing targets was investigated and we show that, although expressing Fas ligand on their surface, LMP2a-specific CD4+ effector cells kill their targets using the Ca2+-dependent perforin/granzyme pathway which is the same mechanism used by CD8+ CTLs.
Su
Zhen
Z
Medizinische Klinik III, Ludwigs-Maximilians-Universität and Institut für Molekulare Immunologie, GSF National Research Centre for Environment and Health, München, Germany.
Peluso
Mario V
MV
Raffegerst
Silke H
SH
Schendel
Dolores J
DJ
Roskrow
Marie A
MA
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000938:Antigen-Presenting Cells / Q000502:physiology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D023201:CD40 Ligand / Q000235:genetics; D002118:Calcium / Q000502:physiology; D003602:Cytotoxicity, Immunologic / Q000276:immunology*; D006801:Humans; D008055:Lipids / Q000008:administration & dosage; D018942:Macrolides; D012313:RNA / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology*; D019014:fas Receptor / Q000502:physiology*
D000900:Anti-Bacterial Agents; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D008055:Lipids; D018942:Macrolides; D014763:Viral Matrix Proteins; D019014:fas Receptor; D023201:CD40 Ligand; D012313:RNA; C036978:concanamycin A; D002118:Calcium