Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10889311 | Journal of Immunological Methods | 2009 | 8 Pages |
Abstract
Development of methods for efficient in vitro stimulation and expansion of peptide specific CD8+ T cells is compelling not only with respect to adoptive T cell therapy but also regarding analysis of T cell responses and search for new immunogenic peptides. In the present study, a new approach to in vitro T cell stimulation was investigated. By use of an antigenic peptide derived from the cytomegalovirus (CMVp) we tested the stimulatory efficacy of recombinant plate bound MHC molecules (PB-MHC), being immobilized in culture plates. A single stimulation of non-adherent peripheral blood mononuclear cells (NA-PBMCs) with PB-MHC/CMVp resulted in significant expansion of CMVp specific CD8+ T cells, which was comparable to that achieved by CMVp pulsed mature dendritic cells (DCs). By repeated exposure of NA-PBMCs to PB-MHC/CMVp more than 60% CMVp specific CD8+ T cells, representing a 240-fold expansion, were reached after only two stimulations. Although stimulation with PB-MHC/CMVp clearly demonstrated efficient peptide specific expansion of CD8+ T cells, there was a tendency to proliferative exhaustion of the cells after 3-4 stimulations. Thus, it will be of interest to examine the effect of new stimulatory cocktails, e.g. cytokines and co-stimulatory molecules, by use of the present rapid and easy-to-use method of expanding peptide specific T cells.
Keywords
Related Topics
Life Sciences
Biochemistry, Genetics and Molecular Biology
Biotechnology
Authors
Esben G.W. Schmidt, Søren Buus, Mette Thorn, Anette Stryhn, Christian Leisner, Mogens H. Claesson,