Multimer Staining of Cytomegalovirus Phosphoprotein 65–Specific T Cells for Diagnosis and Therapeutic Purposes: A Comparative Study

Abstract

These findings demonstrate the power of multimer staining and a certain limitation of serologic testing to define appropriate donors for transplantation. Therefore, whenever possible, CMV-seropositive donors of transplants to seropositive recipients should be screened for their CD8(+) T cell frequency. All 3 multimer technologies can be used, yielding similar results. The streptamer technology additionally offers the advantage of selecting CMV phosphoprotein 65-specific CD8(+) T cells at the good manufacturing practice level for adoptive T cell transfer.

Adoptive transfer of pp65-specific T cells for the treatment of chemorefractory cytomegalovirus disease or reactivation after haploidentical and matched unrelated stem cell transplantation

“…Therefore, a combination of CD4 ϩ and CD8 ϩ T cells for adoptive transfer is supposed to be beneficial to restore a sustained and protective immunity. Although purity is theoretically higher in T cells sorted by peptide major histocompatibility complex multimers, 25 compared with cytokine-capture systems, the isolation of IFN-␥-secreting cells enables the generation of CD4 ϩ and CD8 ϩ T-cell responses to multiple epitopes, 26 the application of functionally active (IFN-␥ secreting) T cells, and applicability to all patients, independent of the HLA type.…”

Immunotherapy for transplantation-associated viral infections

“…There are a number of cell selection reagents available, including tetramers, pentamers, and streptamers. Similar frequencies of multimerpositive T cells can be detected following selection using all three techniques, but tetramers possess the lowest background signal and can potentially deliver the purest product (50).…”

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

“…However, this difference has not increased the sensibility of the technique. In fact, Yao et al 28 did not find any differences in CMV phosphoprotein 65-specific CD8( þ ) T-cell frequencies using tetramers and pentamers in CMV-seropositive healthy volunteers and CMV-seropositive patients before and after allogeneic PBSC transplantation. Tetramers have been evaluated by the European Working Group on Clinical Cell Analysis, and efforts have been made towards a reproducible assay for routine enumeration of antigen-specific CD8 þ T cells.…”

“…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.…”