Induction of Epstein-Barr virus-specific cytotoxic T-lymphocyte responses using dendritic cells pulsed with EBNA-3A peptides or UV-inactivated, recombinant EBNA-3A vaccinia virus.
Cell-mediated immunity, especially the cytotoxic T lymphocyte (CTL), provides resistance to Epstein-Barr virus (EBV), as is demonstrated by the occurrence of posttransplant lymphoproliferative disease in immunosuppressed patients. We set out to use dendritic cells (DCs) to elicit anti-EBV-specific CTLs in culture. In unselected, HLA-B8(+) donors, monocyte-derived mature DCs were pulsed with the HLA-B8-restricted EBNA-3A peptide, FLRGRAYGL, and added to autologous T cells for 7 days at a DC:T ratio of 1:5 to 1:60. The cultured cells specifically lysed EBNA-3A peptide-pulsed, HLA-B8(+), B-lymphoblastoid cell lines in a 5-hour (51)Cr-release assay. The generation of CTLs did not require the addition of interleukin-2. In comparison, monocytes were weak antigen-presenting cells. DCs were then infected with recombinant vaccinia-EBNA-3A. Vaccinia infection significantly decreased the viability of immature DCs after 3 days of culture (to 25% to 45%) but had a smaller effect on mature DC recovery (40% to 70%). To decrease these cytopathic effects and to expand the potential use of vaccinia vectors for DC therapy in immunocompromised patients, we successfully used psoralen and UV-inactivated virus. Mature DCs pulsed with either live or inactivated vaccinia EBNA-3A virus could elicit strong EBNA-3A-specific CTLs. Therefore, mature DCs are powerful stimulators of EBV-specific CTLs and their major histocompatibility complex class I products can even be charged with UV-inactivated recombinant vaccinia.
Subklewe
M
M
Laboratory of Cellular Physiology and Immunology, Rockefeller University, New York, NY, USA. subklem@rockvax.rockefeller.edu
Chahroudi
A
A
Schmaljohn
A
A
Kurilla
M G
MG
Bhardwaj
N
N
Steinman
R M
RM
D017951:Antigen Presentation; D000956:Antigens, Viral / Q000276:immunology*; D003602:Cytotoxicity, Immunologic; D004274:DNA, Recombinant; D003713:Dendritic Cells / Q000276:immunology* / Q000821:virology; D015795:HLA-B8 Antigen / Q000276:immunology; D006801:Humans; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014466:Ultraviolet Rays; D014616:Vaccinia virus / Q000235:genetics / Q000276:immunology* / Q000528:radiation effects; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*
D000956:Antigens, Viral; D004274:DNA, Recombinant; D015795:HLA-B8 Antigen; D014764:Viral Proteins