Reversible HLA multimers (Streptamers) for the isolation of human cytotoxic T lymphocytes functionally active against tumor- and virus-derived antigens

Viral-specific adoptive immunotherapy after allo-SCT: the role of multimer-based selection strategies

“…30 Other authors have observed the unusual functional status of the specific T cells after tetramer selection, given that lytic function and proliferation of the specific T cells was impaired after joining with conventional tetramers. 31 However, this phenomenon of functional disruption has not been observed in vivo conditions. 32 This group found that CMV-specific CTLs selected with tetramer technology and transferred to SCT recipients efficiently contributed to the control of viral replication.…”

“…Following dissociation, the streptamer-treated CTLs are functionally indistinguishable from untreated T cells. 31 Wang et al 35 demonstrated that the in vitro treatment of the T cells with peptide-loaded OT-I-streptamers induced an increased proliferation and strong expression of CD69 and CD25 activation markers. It also induced a sustained phosphorylation of Akt (a serine threonine kinase, which is critically involved in T-cell survival) and ERK1/2 (serine/threonine kinase important for cellular proliferation and survival), a weak activation of caspase-3, a weak anexin V and propidium iodide expression and expression of Bcl-x L (that prevents TCR-mediated apoptosis and promotes T-cell survival).…”

“…Moreover, streptamers have comparable sensitivity and specificity to conventional tetramers and pentamers, 28,31 and the streptamer technology additionally offers the advantage of selecting specific T cells under good manufacturing practice conditions for adoptive T-cell transfer. This later aspect has so far proved a significant challenge for translational researchers.…”

Reversible Major Histocompatibility Complex I-Peptide Multimers Containing Ni2+-Nitrilotriacetic Acid Peptides and Histidine Tags Improve Analysis and Sorting of CD8+ T Cells

“…A major shortcoming of these tetramers is that they avidly bind to and cross-link cell surface TCR and CD8, thereby inducing strong T cell activation, which frequently provokes T cell death (4,5,8,9). Thus, isolation of antigen-specific CD8ϩ T cells by tetramers harbors the risk of substantial T cell loss.…”

“…Thus, isolation of antigen-specific CD8ϩ T cells by tetramers harbors the risk of substantial T cell loss. To circumvent this, "reversible" tetramers have been developed that contain low affinity biotin analogues and therefore dissociate upon addition of free biotin (4,5,9). However, although these reagents improved sorting and cloning of live antigenspecific CD8ϩ T cells, they are costly to produce and of limited stability, even at room temperature, commonly used for FACS sorting.…”