International Immunology, Vol. 12, No. 12, 1741-1747,
December 2000
© 2000 Japanese Society for Immunology
Dendritic cells efficiently cross-prime HLA class I-restricted cytolytic T lymphocytes when pulsed with both apoptotic and necrotic cells but not with soluble cell-derived lysates
1 Istituto Nazionale per la Ricerca sul Cancro, 16132 Genova, Italy
2 Dipartimento di Oncologia, Biologia e Genetica, Università degli Studi di Genova, 161321 Genova, Italy
Correspondence to: G. Ferlazzo
Dendritic cells (DC) have been shown to efficiently present antigen to CD8+ cytolytic T cells (CTL) when pulsed with apoptotic cells as a source of cell-derived antigen. Such cross-priming could not be detected by the use of necrotic cells, while conflicting results have been reported for cell-derived soluble lysates. In this study, we reinvestigated this issue by using autologous Epstein–Barr virus-transformed lymphoblastoid cell lines (LCL) as a source of antigen to pulse monocyte-derived DC. Both autologous and HLA-mismatched allogeneic LCL have been used either in the form of apoptotic or of necrotic cells or of soluble cell lysates. At day 7, DC were co-cultured with the autologous CD8+ lymphocyte fraction for an additional 9 days, in the presence of exogenous IL-2 (added after 48 h). At the end of the culture period, CD8+ CTL efficiently lysed autologous LCL only when they had been co-cultured with DC pulsed with necrotic or apoptotic cells. That an efficient cross-priming of autologous CD8+ cells could be induced by DC pulsed with apoptotic or necrotic LCL but not with cell lysates was further demonstrated in assays of IFN-
production in response to short-term re-stimulation of CD8+ cells with LCL. In addition, LCL-specific CD8+ cells could specifically lyse autologous DC that had been pulsed with LCL-derived antigens, further suggesting that DC presented exogenous antigens on HLA class I molecules.
Keywords: apoptosis, cell lysate, cross-priming, dendritic cell, Epstein–Barr virus-transformed lymphoblastoid cell line
Transmitting editor: L. Moretta
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