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Blood, Vol. 95 No. 4 (February 15), 2000:
pp. 1342-1349
Stimulation of cytotoxic T cells against idiotype immunoglobulin
of malignant lymphoma with protein-pulsed or idiotype-transduced
dendritic cells
Frank Osterroth,
Annette Garbe,
Paul Fisch, and
Hendrik Veelken
From the Departments of Hematology/Oncology and Pathology, Freiburg
University Medical Center; the Department of Biology, Freiburg
University; and Cellgenix GmbH, Freiburg, Germany.
Because of their hypervariable regions and somatic mutations, the
antigen receptor molecules of lymphomas (idiotypes) are tumor-specific antigens and attractive targets for antilymphoma immunotherapy. For the optimal induction of human idiotype-specific cytotoxic T cells (CTL), idiotype was presented to
CD8+ peripheral blood mononuclear cells by
monocyte-derived autologous dendritic cells (DC) after the endocytosis
of idiotype protein or by idiotype-expressing DC. Recombinant
idiotype was obtained as a functionally folded Fab fragment by
periplasmic expression in Escherichia coli. Idiotype-expressing
DC were generated by transduction with recombinant Semliki forest virus
vectors encompassing heavy- or light-chain idiotype genes.
Autologous lymphoblastoid cell lines stably transfected with
Epstein-Barr virus-based idiotype expression vectors were used as
target cells to detect idiotype-specific lysis. CTL stimulated with
idiotype-loaded DC showed strong specific, CD8-mediated, and major
histocompatibility complex (MHC) class I-restricted cytotoxicity
against autologous heavy- and light-chain idiotype. In contrast,
stimulation with idiotype-transduced DC resulted in only moderate
natural killer cell activity. These data confirm the existence of
idiotype-specific CTL in patients with lymphoma, define a "good
manufacturing practice"-compatible protocol for the generation of
these cells without the requirement of viable lymphoma cells, and favor
the processing of exogenous antigen over DC transduction for the
induction of MHC I-restricted CTL against idiotypes with unknown antigenicity.
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