Supplementary MaterialsSupplemental data JCI0832103sd. cells to increase or even to engineer

Supplementary MaterialsSupplemental data JCI0832103sd. cells to increase or even to engineer for adoptive immunotherapy of human being infections or malignancy. Introduction Studies in rodents have demonstrated that adoptive immunotherapy with antigen-specific CD8+ cytotoxic T cells is effective for cancer and infections, and there is evidence that this approach has therapeutic activity in humans (1C8). For clinical applications, T cells of a desired antigen specificity are isolated or engineered to express receptors that target infected or transformed cells and are then expanded in culture (9C14). In some settings the transfer of cloned T cells has Rabbit polyclonal to Noggin been used to provide precise control of specificity and avoid toxicity. For example, in allogeneic stem cell transplantation, the administration of donor-derived T cell clones that target pathogens or malignant Vistide inhibitor database cells in the recipient can avoid graft-versus-host disease, which occurs with the infusion of unselected polyclonal donor T cells (3, 4, 15). However, the efficacy of adoptive immunotherapy in humans is usually often limited by the failure of cultured T cells, particularly cloned CD8+ T cells, to persist in vivo (16, 17), and insight into the basis for the poor survival of the transferred cells is usually lacking. The pool of lymphocytes from which CD8+ T cells for adoptive immunotherapy can be derived includes naive T cells (TN) and antigen-experienced memory T cells (TM), which can be divided into central memory (TCM) and effector memory (TEM) subsets that differ in phenotype, homing, and function (18). CD8+ TCM express CD62L and CCR7, which promote migration into LNs and proliferate rapidly if reexposed to antigen (19). CD8+ TEM lack CD62L, enabling migration to peripheral tissues, and exhibit immediate effector function (19). In response to antigen stimulation, both CD8+ TCM and TEM proliferate and differentiate into CD62LC cytolytic effector T cells (TE) that express high levels of granzymes and perforin Vistide inhibitor database but are short lived (20). Thus acquisition of an effector phenotype during culture has been suggested as a major reason for the poor survival of moved T cells (9). In the standard web host, T cell storage persists forever, indicating that some TM cells may be capable of self-renew or revert towards the storage pool after differentiating to TE in response to repeated antigen publicity (21). TEM and TCM possess specific phenotypic and useful properties, but it is certainly unidentified whether TE cells produced from each one of these TM subsets retain any intrinsic properties from the parental cell. Utilizing a non-human primate model highly relevant to human translation, we sought to determine whether TE clones derived from purified TCM or TEM differed Vistide inhibitor database in their ability to persist in vivo or establish T cell memory after adoptive transfer. Here we show that antigen-specific CD8+ TE clones derived from the TEM subset of TM survive in the blood for only a short period after adoptive transfer, fail to home to LNs or BM, and do not reacquire phenotypic markers of TM. By contrast, TE clones derived from TCM persist long term after adoptive transfer, migrate to TM niches, reacquire phenotypic properties of TM, and respond to antigen challenge. Results Characterization of CMV-specific CD8+ T cell clones from CD62L+ TCM and CD62LC Vistide inhibitor database TEM subsets. Immunocompetent with latent CMV contamination were used in this study. We recognized CMV epitopes recognized by CD8+ T cells in individual macaques by stimulating aliquots of PBMCs with CMV immediate early 1 (IE-1) or IE-2 peptides and analyzing IFN- production by circulation cytometry (22). We then determined whether the CD8+ T cells that made IFN- after CMV activation were present in TCM, TEM, and/or TN subsets using cytokine circulation cytometry after staining with CD8-, CD28-, and CD95 (Fas)Cspecific mAbs. TN and TCM are both CD62L+ and CD28+ but can be distinguished from each other by differential expression of Fas,.

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