Peptide specific expansion of CD8 super(+) T cells by recombinant plate bound MHC/peptide complexes

• Published in: Journal of immunological methods Vol. 340; no. 1; pp. 25 - 32
• Main Authors: Schmidt, EGW, Buus, S, Thorn, M, Stryhn, A, Leisner, C, Claesson, M H
• Format: Journal Article
• Language: English
• Published: 01.01.2009
• Subjects: Cytomegalovirus
• ISSN: 0022-1759
• DOI: 10.1016/j.jim.2008.09.020

Development of methods for efficient in vitro stimulation and expansion of peptide specific CD8 super(+) 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 super(+) 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 super(+) 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 super(+) 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.