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Human peripheral blood monuclear cells transfected with messenger RNA stimulate antigen-specific cytotoxic T-lymphocytes in vitro
Authors:R. Teufel  J.-P. Carralot  B. Scheel  J. Probst  S. Walter  G. Jung  I. Hoerr  H.-G. Rammensee  S. Pascolo
Affiliation:(1) CureVac GmbH, Paul-Ehrlich-Str. 15, 72076 Tübingen, Germany;(2) Department of Immunology, Institute for Cell Biology, Auf der Morgenstelle 15, 72076 Tübingen, Germany;(3) Institute for Organic Chemistry, Auf der Morgenstelle 18, 72076 Tübingen, Germany
Abstract:
The efficiency of test vaccines needs to be evaluated by quantification of the triggered cellular immune response. Usually, for these assays, autologous target cells expressing the vaccine antigen are required. In the context of messenger RNA (mRNA)-based vaccinations, the target cells used for the read-out are mRNA-transfected monocyte-derived dendritic cells (Mo-DCs). Their production typically requires samples of 100 ml blood from the patients, and limits the number of assays that can be performed. We show here that fresh peripheral blood mononuclear cells (PBMCs) can be transfected with mRNA by electroporation. Such cells are as efficient as mRNA-transfected Mo-DCs for their ability to activate memory T cells in vitro. Thus, mRNA-transfected PBMCs are a convenient replacement of mRNA-transfected Mo-DCs for the in vitro monitoring of natural or vaccine-induced immune responses.Received 17 February 2005; received after revision 1 May 2005; accepted 7 Juni 2005
Keywords:mRNA  B lymphocyte  dendritic cell  Monocyte  T lymphocyte  MHC tetramer
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