Robust Identification of Suitable T-Cell Subsets for Personalized CMV-Specific T-Cell Immunotherapy Using CD45RA and CD62L Microbeads

Abstract

Viral infections and reactivations remain a serious obstacle to successful hematopoietic stem cell transplantation (HSCT). When antiviral drug treatment fails, adoptive virus-specific T-cell transfer provides an effective alternative. Assuming that naive T cells (TN) are mainly responsible for GvHD, methods were developed to generate naive T-cell-depleted products while preserving immune memory against viral infections. We compared two major strategies to deplete potentially alloreactive T cells: CD45RA and CD62L depletion and analyzed phenotype and functionality of the resulting CD45RA−/CD62L− naive T-cell-depleted as well as CD45RA+/CD62L+ naive T-cell-enriched fractions in the CMV pp65 and IE1 antigen model. CD45RA depletion resulted in loss of terminally differentiated effector memory T cells re-expressing CD45RA (TEMRA), and CD62L depletion in loss of central memory T cells (TCM). Based on these differences in target cell-dependent and target cell-independent assays, antigen-specific T-cell responses in CD62L-depleted fraction were consistently 3–5 fold higher than those in CD45RA-depleted fraction. Interestingly, we also observed high donor variability in the CD45RA-depleted fraction, resulting in a substantial loss of immune memory. Accordingly, we identified donors with expected response (DER) and unexpected response (DUR). Taken together, our results showed that a naive T-cell depletion method should be chosen individually, based on the immunophenotypic composition of the T-cell populations present.

Reappearance of Effector T Cells Predicts Successful Recovery from COVID-19

“…To characterize the involvement of different subsets of αβ CD4 conv , αβ CD8 + , and γδ T cells based on their antigen experience 18,19 , we developed a staining panel dedicated to identifying four distinct populations based on CD62L and CD45RA expression ( Supplementary Fig. 2B ).…”

Novel immunological approach to asses donor reactivity of transplant recipients using a humanized mouse model

“…A recent report on adoptive virus-specific T cell transfer showed that the CD45RA-depleted population with loss of TEMRA cells substantially lost donor-specific immune responses. In contrast, the CD62L-depleted population with loss of TCM cells consistently yielded 3-5 folds higher donor Ag responses [44]. In addition, it has been reported that the main driving force behind AR is TEM cells in the transplant field [45].…”

Antiviral T-Cell Frequencies in a Healthy Population: Reference Values for Evaluating Antiviral Immune Cell Profiles in Immunocompromised Patients

“…IFN-γ ELISpot assay was performed as described previously [44,45]; details are provided in supplementary information (Methods, Tab.S3).…”