C-Terminal Incorporation of &#945-Trifluoromethyl Substituted Amino Acids into Peptides via <i>in situ</i>-Deprotection of N-Teoc-Derivatives

Hollweck W, Sewald N, Michel T, Burger K. C-Terminal Incorporation of &amp;#945-Trifluoromethyl Substituted Amino Acids into Peptides via &lt;i&gt;in situ&lt;/i&gt;-Deprotection of N-Teoc-Derivatives. Liebigs Ann./Recueil. 1997;1997(12):2549-2551

Peptide modification by introduction of alpha-trifluoromethyl substituted amino acids

Sewald N, Hollweck W, Mütze K, et al. Peptide modification by introduction of alpha-trifluoromethyl substituted amino acids. Amino Acids. 1995;8(2):187-194

B cells immortalized by a mini-Epstein-Barr virus encoding a foreign antigen efficiently reactive specific cytotoxic T cells.

Lymphoblastoid cell lines (LCLs) are human B cells latently infected and immortalized by Epstein-Barr virus (EBV). Presenting viral antigens, they efficiently induce EBV-specific T-cell responses in vitro. Analogous ways to generate T-cell cultures specific for other antigens of interest are highly desirable. Previously, we constructed a mini-EBV plasmid that consists of less than half the EBV genome, is unable to cause virus production, but still immortalizes B cells in vitro. Mini-EBV&ndash;immortalized B-cell lines (mini-LCLs) are efficiently produced by infection of B cells with viruslike particles carrying only mini-EBV DNA. Mini-EBV plasmids can be engineered to express an additional gene in immortalized B cells. Here we present a mini-EBV coding for a potent CD8+ T-cell antigen, the matrix phosphoprotein pp65 of human cytomegalovirus (CMV). By means of this pp65 mini-EBV, pp65-expressing mini-LCLs could be readily established from healthy donors in a one-step procedure. We used these pp65 mini-LCLs to reactivate and expand effector T cells from autologous peripheral blood cells in vitro. When generated from cytomegalovirus (CMV)&ndash;seropositive donors, these effector T-cell cultures displayed strong pp65-specific HLA-restricted cytotoxicity. A large fraction of CD8+ T cells with pp65 epitope specificity was present in such cultures, as demonstrated by direct staining with HLA/peptide tetramers. We conclude that the pp65 mini-EBV is an attractive tool for CMV-specific adoptive immunotherapy. Mini-EBVs could also facilitate the generation of T cells specific for various other antigens of interest